BIOMARKERS FOR COLORECTAL CANCER

BACKGROUND

Colorectal cancer (CRC) is the second most deadly cancer, causing approximately 500,000 deaths per year. Early detection is paramount to reducing the mortality associated with this disease. Yet, present screening methods are less than effective. Furthermore, present methods are time-consuming, costly and inconvenient for patients.

Colonoscopy is the most commonly used screening method. A variety of factors limit the effectiveness of this method, however. For example, changes in the colon are sometimes invisible during a colonoscopy, and biopsies must be taken during the procedure and examined under a microscope to detect cancerous or precancerous changes. Also, the method's accuracy depends on the experience of the practitioner. Thus, results from different colonoscopies can vary and precancerous changes may go undetected. Colonoscopy also causes patient discomfort and carries certain risks, such as bleeding or puncture of the lining of the colon.

Thus, there is a need for a quick, reliable, non-invasive test for colorectal cancer.

SUMMARY

In one embodiment, a method for detecting colorectal cancer in a patient comprises obtaining a biological sample from the patient and evaluating the sample or a fraction of the sample for the presence of at least one biomarker selected from the group of peptides having the sequences of SEQ ID NOs: 1-388, wherein the presence of said at least one biomarker is indicative of colorectal cancer. In another, the methods involve evaluating the sample for the presence of a biomarker selected from the group of peptides having the amino acid sequence of SEQ ID NOs: 176-388. In another embodiment, the methods comprise evaluating the sample for the presence of peptides having the amino acid sequence of SEQ ID NOs: 176, 177, and 234.

In one aspect, the colorectal cancer is in early stage, such as stage T1 or T2. The biological sample can be, for example, blood, serum or plasma. In another, the evaluation step comprises assays such as mass spectrometry, an immunoassay such as ELISA, immuno-mass spectrometry or suspension bead array.

In another embodiment, the method further comprises, prior to the evaluation step, harvesting low molecular weight peptides from the biological sample to generate at least one fraction comprising the peptides. In one embodiment, the size of the low molecular weight peptides is less than 50 KDa, preferably less than 25 KDa, and more preferably less than 15 KDa. In another aspect, the method also comprises digesting the low molecular weight peptides. Such digestion can be accomplished using enzymatic or chemical means. In one example, trypsin can be used to digest the peptides.

In another aspect, a method for monitoring the progression of colorectal cancer in a patient comprises (i) obtaining a biological sample from the patient, (ii) evaluating the sample or a fraction of the sample for the presence of at least one biomarker selected from the group of peptides having the sequences of SEQ ID NOs: 1-388, wherein the presence of said at least one biomarker is indicative of colorectal cancer, and optionally, repeating steps (i) and (ii) as necessary. In another, the methods involve evaluating the sample for the presence of a biomarker selected from the group of peptides having the amino acid sequence of SEQ ID NOs: 176-388. In another embodiment, the methods comprise evaluating the sample for the presence of peptides having the amino acid sequence of SEQ ID NOs: 176, 177, and 234. In one embodiment, the method further comprises a step of harvesting low molecular weight peptides from the sample to generate at least one fraction comprising the peptides.

In other aspects, the invention relates to antibodies specific for identified biomarkers for colorectal cancer, as well as kits for detecting colorectal cancer in a patient, comprising at least one such antibody.

Other objects, features and advantages will become apparent from the following detailed description. The detailed description and specific examples are given for illustration only since various changes and modifications within the spirit and scope of the invention will become apparent to those skilled in the art from this detailed description. Further, the examples demonstrate the principle of the invention and cannot be expected to specifically illustrate the application of this invention to all the examples where it will be obviously useful to those skilled in the prior art.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 provides a CID Spectrum of peptide “TFSLSSTLLR” from Zinc finger protein 553 identified only in LMW of colorectal cancer serum (accession number Q4G0R3, amino acid residues 367-376).

FIG. 2 provides a CID Spectrum of peptide “DMKPENLLCMGPELVK” from Serine/threonine-protein kinase MAK identified only in LMW of colorectal cancer serum (accession number P20794, amino acid residues 125-140).

FIG. 3 provides a CID Spectrum of peptide “SGVQQLIQYYQDQK” from Apolipoprotein F precursor identified only in LMW of colorectal cancer serum (accession number Q13790, amino acid residues 233-246).

DETAILED DESCRIPTION

Low molecular weight (LMW) peptides have been discovered that are indicative of colorectal cancer. Evaluating patient samples for the presence of such LMW peptides is an effective means of detecting colorectal cancer and monitoring the progression of the disease, for example during treatment. The LMW peptides are particularly useful in detecting colorectal cancer during its early stages.

The LMW peptides, or biomarkers, can be detected using a variety of methods known in the art. For example, antibodies can be utilized in immunoassays to detect the presence of a biomarker. Exemplary immunoassays include, e.g., ELISA, radioimmunoassay, immunofluorescent assay, “sandwich” immunoassay, western blot, immunoprecipitation assay and immunoelectrophoresis assays. In other examples, beads, microbeads, arrays, microarrays, etc. can be applied in detecting the LMW peptides. Exemplary assays that can be used include suspension bead assays (Schwenk et al., “Determination of binding specificities in highly multiplexed bead-based assays for antibody proteomics,” Mol. Cell Proteomics, 6(1): 125-132 (2007)), antibody microarrays (Borrebaeck et al., “High-throughput proteomics using antibody microarrays: an update,” Expert Rev. Mol. Diagn. 7(5): 673-686 (2007)), aptamer arrays (Walter et al., “High-throughput protein arrays: prospects for molecular diagnostics,” Trends Mol. Med. 8(6): 250-253 (2002)), affybody arrays (Renberg et al., “Affibody molecules in protein capture microarrays: evaluation of multidomain ligands and different detection formats,” J. Proteome Res. 6(1): 171-179 (2007)), and reverse phase arrays (VanMeter et al., “Reverse-phase protein microarrays: application to biomarker discorvery and translational medicine,” Expert Rev. Mol. Diagn. 7(5): 625-633 (2007)). The referenced publications are hereby incorporated by reference.

In another example, the inventive biomarkers can be detected using mass spectrometry (MS). One example of this approach is tandem mass spectrometry (MS/MS), which involves multiple steps of mass selection or analysis, usually separated by some form of fragmentation. Most such assays use electrospray ionization followed by two stages of mass selection: a first stage (MS1) selecting the mass of the intact analyte (parent ion) and, after fragmentation of the parent by collision with gas atoms, a second stage (MS2) selecting a specific fragment of the parent, collectively generating a selected reaction monitoring assay. In one embodiment, collision-induced dissociation is used to generate a set of fragments from a specific peptide ion. The fragmentation process primarily gives rise to cleavage products that break along peptide bonds. Because of the simplicity in fragmentation, the observed fragment masses can be compared to a database of predicted masses for known peptide sequences. A number of different algorithmic approaches have been described to identify peptides and proteins from tandem mass spectrometry (MS/MS) data, including peptide fragment fingerprinting (SEQUEST, MASCOT, OMSSA and X!Tandem), peptide de novo sequencing (PEAKS, LuteFisk and Sherenga) and sequence tag based searching (SPIDER, GutenTAG).

Likewise, multiple reaction monitoring (MRM) can be used to identify the inventive biomarkers in patient samples. This technique applies the MS/MS approach to, for example, tryptic digests of the input sample, followed by selected ion partitioning and sampling using MS to objectify and discreetize the analyte if interest by following the exact m/z ion of the tryptic fragment that represents the analyte. Such an approach can be performed in multiplex so that multiple ions can be measured at once, providing an antibody-free method for analyte measurement. See, e.g. Andersen et al., “Quantitative mass spectrometric multiple reaction monitoring assays for major plasma proteins,” Molecular & Cellular Proteomics, 5.4: 573-588 (2006); Whiteaker et al., “Integrated pipeline for mass spectrometry-based discorvery and confirmation of biomarkers demonstrated in a mouse model of breast cancer,” J. Proteome Res. 6(10): 3962-75 (2007). Both publications are incorporated herein by reference.

In another example, the inventive biomarkers can be detected using nanoflow reverse-phase liquid chromatography-tandem mass spectrometry. See, e.g., Domon B, Aebersold R. “Mass spectrometry and protein analysis.” Science, 312(5771):212-7(2006), which is incorporated herein by reference in its entirety. Using this approach, experimentalists obtain peptide fragments, usually by trypsin digest, and generate mass spectrograms of the fragments, which are then compared to a database, such as SEQUEST, for protein identification.

In another aspect, the inventive biomarkers can be detected using immuno-mass spectrometry. See, e.g., Liotta L et al. “Serum peptidome for cancer detection: spinning biologic trash into diagnostic gold.” J Clin Invest.,116(1):26-30 (2006); Nedelkov, “Mass spectrometry-based immunoassays for the next phase of clinical applications,” Expert Rev. Proteomics, 3(6): 631-640 (2006), which are hereby incorporated by reference. Immuno-mass spectrometry provides a means for rapidly determining the exact size and identity of a peptide biomarker isoform present within a patient sample. When developed as a high throughput diagnostic assay, a drop of patient's blood, serum or plasma can be applied to a high density matrix of microcolumns or microwells filled with a composite substratum containing immobilized polyclonal antibodies, directed against the peptide marker. All isoforms of the peptide that contain the epitope are captured. The captured population of analytes including the analyte fragments are eluted and analyzed directly by a mass spectrometer such as MALDI-TOF MS. The presence of the specific peptide biomarker at its exact mass/charge (m/z) location would be used as a diagnostic test result. The analysis can be performed rapidly by simple software that determines if a series of ion peaks are present at defined m/z locations.

In yet another example, the inventive biomarkers can be detected using standard immunoassay-based approaches whereby fragment specific antibodies are used to measure and record the presence of the diagnostic fragments. See, e.g., Naya et al. “Evaluation of precursor prostate-specific antigen isoform ratios in the detection of prostate cancer.” Urol Oncol. 23(1):16-21 (2005). Moreover, additional well known immunoassays, such as ELISAs (Maeda et al., “Blood tests for asbestos-related mesothelioma,” Oncology 71: 26-31 (2006)), microfluidic ELISAs (Lee et al., “Microfluidic enzyme-linked immunosorbent assay technology,” Adv. Clin. Chem. 42: 255-259 (2006)), nanocantilever immunoassays (Kurosawa et al., “Quartz crystal microbalance immunosensors for environmental monitoring,” Biosens Bioelectron, 22(4): 473-481 (2006)), and plasmon resonance immunoassays (Nedelkov, “Development of surface Plasmon resonance mass spectrometry array platform,” Anal. Chem. 79(15): 5987-5990 (2007)) can be employed. The referenced publications are hereby incorporated by reference.

In a further example, the inventive biomarkers can be detected using electrochemical approaches. See, e.g., Lin et al., “Electrochemical immunosensor for carcinoembryonic antigen based on antigen immobilization in gold nanoparticles modified chitosan membrane,” Anal. Sci. 23(9): 1059-1063 (2007).

In one embodiment, the LMW peptides are harvested from a biological sample prior to the evaluation step. For example, 100 μl of serum can be mixed with 2×SDS-PAGE Laemmli Buffer (containing 200 mM DTT), boiled for 10 minutes, and loaded on Prep Cell (Model 491 Prep Cell, Bio-Rad Laboratories, CA) comprising a 5 cm length 10% acrylamide gel. Electrophoresis is performed under a constant voltage of 250V. Immediately after the bromophenol blue indicator dye is eluted from the system, LMW peptides and proteins migrate out of the gel and are trapped in a dialysis membrane in the elution chamber. These molecules can be eluted at a flow rate of 400 ml/min by a buffer with the same composition of the Tris-Glycine running buffer and collected for 10 minutes in one fraction.

Alternatively, LMW peptides can be harvested using from a sample using a capture-particle that comprises a molecular sieve portion and an analyte binding portion as described in U.S. patent application Ser. No. 11/527,727, filed Sep. 27, 2006, which is hereby incorporated by reference. Briefly, either the molecular sieve portion or the analyte binding portion or both comprise a cross-linked region having modified porosity, or pore dimensions sufficient to exclude high molecular weight molecules.

In another embodiment, the LMW peptides are digested prior to detection, so as to reduce the size of the peptides. Such digestion can be carried out using standard methods well known in the field. Exemplary treatments, include but are not limited to, enzymatic and chemical treatments. Such treatments can yield partial as well as complete digestions. One example of an enzymatic treatment is a trypsin digestion.

The inventive biomarkers are particularly useful in detecting colorectal cancer during its early stages, i.e., prior to metastasis and large tumor volume (e.g. greater than 2 cm).

Antibodies specific for the inventive biomarkers can be produced readily using well known methods in the art. See, e.g. J. Sambrook, E. F. Fritsch and T. Maniatis, Molecular Cloning, a Laboratory Manual, second edition, Cold Spring Harbor Laboratory Press, pp. 18.7-18.18, 1989). For example, the inventive biomarkers can be prepared readily using an automated peptide synthesizer. Next, injection of an immunogen, such as (peptide)_n-KLH (n=1-30) in complete Freund's adjuvant, followed by two subsequent injections of the same immunogen suspended in incomplete Freund's adjuvant into immunocompetent animals, is followed three days after an i.v. boost of antigen, by spleen cell harvesting. Harvested spleen cells are then fused with Sp2/0-Ag14 myeloma cells and culture supernatants of the resulting clones analyzed for anti-peptide reactivity using a direct-binding ELISA. Fine specificity of generated antibodies can be detected by using peptide fragments of the original immunogen.

In certain embodiments, one or more antibodies directed to the inventive biomarkers is provided in a kit, for use in a diagnostic method. Such kits also can comprise reagents, instructions and other products for performing the diagnostic method.

Examples
Example 1
Identification of Biomarkers for Colorectal Cancer

Blood Collection and Serum Preparation

Blood samples were drawn from patients before the colonoscopy test under full Institutional Review Board approval and patient's consent. Specimens were collected in red-top Vacutainer Tubes and allowed to clot for 1 hour on ice, followed by centrifugation at 4° C. for 10 minutes at 2000 g. The serum supernatant was divided in aliquots and stored in −80° C. until needed. 10 serum samples with negative outcome were pooled in a single control group. 10 serum samples from patients with a diagnosed T1 or T2 stage colorectal cancer were pooled in a single disease group. Each experiment has been performed using 3 different aliquots from the same pool, both for the control and for the disease group.

Low Molecular Weight (LMW) Protein Harvesting by Continuous Elution Electrophoresis

100 μl of serum was mixed with 2×SDS-PAGE Laemmli Buffer (containing 200 mM DTT), boiled for 10 minutes, and loaded on Prep Cell (Model 491 Prep Cell, Bio-Rad Laboratories, CA) in which 5 cm length 10% acrylamide gel was polymerized. Electrophoresis was performed under a constant voltage of 250V. Immediately after the bromophenol blue indicator dye was eluted from the system, LMW peptides and proteins migrate out of the gel and are trapped in a dialysis membrane in the elution chamber. These molecules were eluted at a flow rate of 400 μl/min by a buffer with the same composition of the Tris-Glycine running buffer and collected for 10 minutes in one fraction.

SOS Removal from the Prep Cell Fractions

LMW fractions obtained by the Prep Cell were processed using a commercially available ion-exchange matrix (Proteo Spin Detergent Clean-Up Micro Kit, Norgen Biotek Corporation, Canada) following protocols outlined by the manufacturer for both acidic and basic proteins, resulting in a final volume of 55 μl.

Nanoflow Reversed-Phase Liquid Chromatography-Tandem MS (nanoRPLC-MS/MS)

The SDS-free LMW fractions obtained from the described procedure were analyzed by traditional bottom-up MS approaches. This was accomplished by treating the samples by reduction using 20 mM DTT, followed by alkylation using 100 mM iodoacetamide and lastly, trypsin digestion (Promega, Wis.) at 37° C. overnight in 50 mM ammonium bicarbonate in the presence of 1M urea in a final volume of 200 μl. Tryptic peptides were desalted by μC₁₈Zip Tip (Millipore, Mass.) and analyzed by reversed-phase liquid chromatography nanospray tandem mass spectrometry using a linear ion-trap mass spectrometer (LTQ, ThermoElectron, San Jose, Calif.). Reverse phase column was slurry-packed in-house with 5 μm, 200 Å pore size C₁₈resin (Michrom BioResources, CA) in 100 μm i.d.×10 cm long fused silica capillary (Polymicro Technologies, Phoenix, Ariz.) with a laser-pulled tip. After sample injection, the column was washed for 5 min with mobile phase A (0.4% acetic acid, 0.005% heptafluorobutyric acid) and peptides were eluted using a linear gradient of 0% mobile phase B (0.4% acetic acid, 0.005% heptafluorobutyric acid, 80% acetonitrile) to 50% mobile phase B in 30 min at 250 nl/min, then to 100% B in an additional 5 min. The LTQ mass spectrometer was operated in a data-dependent mode in which each full MS scan was followed by five MS/MS scans where the five most abundant molecular ions were dynamically selected and fragmented by collision-induced dissociation (CID) using a normalized collision energy of 35%.

Bioinformatic Analysis

Tandem mass spectra were matched against Swiss-Prot human protein database through SEQUEST algorithm incorporated in Bioworks software (version 3.2, Thermo Electron) using tryptic cleavage constraints and static cysteine alkylation by iodoacetamide. For a peptide to be considered legitimately identified, it had to achieve Delta Cn value above 0.1, cross correlation scores of 1.5 for [M+H]¹⁺, 2.0 for [M+2H]²⁺, 2.5 for [M+3H]³⁺, and a probability cut-off for randomized identification of p<0.01.

The results are provided in Table 1. In short, 175 peptides were identified as biomarkers that correlate to the disease state. Thus, evaluating patient samples for the presence of one or more of these biomarkers will provide a useful method for detecting colorectal cancer.

TABLE 1

SEQ ID
Residue

Proteins
P (pro)
MW
Accession
Amino acid sequence
NO.
number

SOCS2_HUMAN (O14508)
8.25E-03
22158.33
O14508
TGPEAPRNGTVHLYLTKPLYTSAP
1
138-168

Suppressor of cytokine

SLQHLCR

signaling 2 (SOCS-2)

(Cytokine-inducible SH2 protein

2)

PGS1_HUMAN (P21810)
5.33E-03
41627.58
P21810
VPSGLPDLKLLQVVYLHSNNITK
2
292-314

Biglycan precursor

(Bone/cartilage proteoglycan I)

(PG-S1)

KCC4_HUMAN (Q16566)
4.36E-03
51892.92
Q16566
IVEKGYYSERDAADAVK
3
130-146

Calcium/calmodulin-dependent

protein kinase type IV (EC

2.7.1.123) (CAM kinase

ZF260_HUMAN (Q3ZCT1) Zinc
9.82E-03
46890.02
Q3ZCT1
HHTGEKPYECEK
4
101-112

finger protein 260 homolog

(Zfp-260)

GP107_HUMAN (Q5VW38)
8.28E-03
66948.05
Q5VW38
KQSVSVTLLILDISR
5
113-127

Protein GPR107 precursor

(Lung seven transmembrane

receptor 1)

Q6UVY4 (Q6UVY4)
4.18E-03
65025.7
Q6UVY4
NYIILNMTENIDCEVFRQHR
6
390-409

Podocalyxin-like protein

ELOA2_HUMAN (Q8IYF1)
2.13E-03
83896.48
Q8IYF1
YLTYDQLRKQK
7
386-396

RNA polymerase II transcription

factor SIll subunit A2 (Elongin

A2) (EloA2) (T

ZN462_HUMAN (Q96JM2) Zinc
6.58E-03
161426.3
Q96JM2
SLLENAEAK
8
1402-1410

finger protein 462

UDB28_HUMAN (Q9BY64)
2.13E-03
60866.27
Q9BY64
ENVMKLSIIQHDQPVK
9
440-455

UDP-glucuronosyltransferase

2B28 precursor (EC 2.4.1.17)

(UDPGT)

A1ATR_HUMAN (P20848)
7.51E-03
47860.97
P20848
TEILEGLNVNLTETPEAK
10
100-117

Alpha-1-antitrypsin-related

protein precursor

Q52LR0 (QS2LR0) Cingulin-like
4.48E-03
149014
Q52LR0
RQVEEAEEEIDRLESSK
11
1213-1229

1

Q5J915 (Q5J915) Migration-
5.97E-03
8760.694
Q5J915
RGKPEILELEGLILLK
12
37-52

inducing protein 7

Q8WWY8 (Q8WWY8)
1.83E-03
50826.5
Q8WWY8
INHEPTTFQK
13
363-372

Membrane-bound phosphatidic

acid-selective phospholipase

A1 (Lipase H) (Lipase, mem

MUCEN_HUMAN (Q9ULC0)
1.22E-03
27435.32
Q9ULCO
TISHESGEHSAQGKTK
14
245-260

Endomucin precursor

(Endomucin-2) (Gastric cancer

antigen Ga34)

NALOL_HUMAN (Q9UQQ1) N-
4.75E-04
80574.27
Q9UQQ1
YVLYGNHRDSWVHGAVDPSSGTAV
15
362-391

acetylated-alpha-linked acidic

LLELSR

dipeptidase-like protein (EC

3.4.17.21) (NAAL

PEPL_HUMAN (O60437)
7.80E-04
204524.7
O60437
QLENEVKSTQEEIWTLR
16
908-924

Periplakin (195 kDa cornified

envelope precursor protein)

(190 kDa paraneoplast

ABLM3_HUMAN (O94929)
1.26E-03
77751.66
O94929
EEIKHGQSLLALDK
17
156-169

Actin-binding LIM protein 3

(Actin-binding LIM protein

family member 3) (abLIM

Q12800 (Q12800) Transcription
6.35E-03
57276.84
Q12800
ANPTQLNTVEFLWDPAK
18
165-181

factor LSF

Q6PJ08 (Q6PJ08) SLC29A4
6.04E-04
56365.03
Q6PJ08
DSPAHEVTGSGGAYMRFDVPR
19
295-315

protein

Q6ZPC4 (Q6ZPC4)
4.13E-03
13386.64
Q6ZPC4
NDPLLGMCSEMGFSGVRESFQR
20
102-123

Hypothetical protein FLJ26077

Q8IY84 (Q8IY84) Hypothetical
9.65E-03
49574.63
Q81Y84
SILEGTYSVPPHVSEPCHRLIR
21
282-303

protein MGC42105

TR137_HUMAN (O94972)
1.34E-04
107837.5
O94972
YEYRVEMVHQSCNDPTKNIIR
22
341-361

Tripartite motif protein 37

(Mulibrey nanism protein)

Q495G5 (Q495G5) MMAA
1.09E-03
47105.97
Q495G5
VGLSGPPGAGKSTFIEYFGK
23
146-165

protein

Q4G0R3 (Q4G0R3) Zinc finger
8.23E-03
67736.06
Q4GOR3
TFSLSSTLLR
24
367-376

protein 553

Q659A1 (Q659A1) Hypothetical
6.70E-03
109923.9
Q659A1
FMMSKNPSVPVSAVFMDK
25
349-366

protein DKFZp31 312410

Z183L_HUMAN (Q8IZP6) Zinc
5.85E-03
36235.56
Q8IZP6
ETGFCGFGDSCKFLHDR
26
201-217

finger protein 183-like 1 (RING

finger protein 161)

Q8TEE6 (Q8TEE6) FLJ00251
2.44E-03
110414.6
Q8TEE6
AAGQCNNMGQKR
27
754-765

protein (Fragment)

PRP18_HUMAN (Q99633) Pre-
1.63E-03
39834.98
Q99633
ESITDIIKFMLQR
28
248-260

mRNA splicing factor 18

(PRP18 homolog) (hPRP18)

UBP21_HUMAN (Q9UK80)
9.83E-03
62616.42
Q9UK80
TSGPRPRGPLR
29
73-83

Ubiquitin carboxyl-terminal

hydrolase 21 (EC 3.1.2.15)

(Ubiguitin_thiolesteras

PSD3_HUMAN (O43242) 26S
2.65E-03
60939.61
O43242
EREQQDLEFAKEMAEDDDDSFP
30
513-534

proteasome non-ATPase

regulatory subunit 3 (26S

proteasome regulatory subun

MYG_HUMAN (P02144)
2.18E-03
17041.91
P02144
HGATVLTALGGILK
31
64-77

Myoglobin

ITPR3 HUMAN (014573)
9.83E-04
303842.3
Q14573
NVRSMPLIYWFSR
32
2187-2199

Inositol 1,4,5-trisphosphate

receptor type 3 (Type 3 inositol

1,4,5-trisphosph

ESPL1_HUMAN (Q14674)
1.10E-03
232963.8
Q14674
GVSLLLSVLRDPALQK
33
877-892

Separin (EC 3.4.22.49)

(Separase) (Caspase-like

protein ESPL1) (Extra spindle

Q573B4 (Q573B4) Proto-
7.31E-03
58296.41
Q573B4
HYTRYYNGHTK
34
266-276

oncogene tyrosine-protein

kinase LCK

DAAM2_HUMAN (Q86T65)
9.87E-03
123420.4
Q86T65
RGLRAVEVELEYQR
35
886-899

Disheveled associated activator

of morphogenesis 2

VCIP1_HUMAN (Q96JH7)
4.82E-03
134235.9
Q96JH7
CLLCGALSELHVPPEWLAPGGK
36
469-490

Deubiquitinating protein

VC1P135 (EC 3.4.22.-)

(Valosin-containing protein p97

THYG_HUMAN (P01266)
6.86E-03
304534.4
P01266
RFLAVQSVISGR
37
283-294

Thyroglobulin precursor

Q8IXT1 (Q8IXT1) Hypothetical
7.64E-03
111478.8
Q8IXT1
ESDHSSLNNK
38
566-575

protein FLJ25416

(Q9BVY9) ZNF160 protein
8.21E-03
17100.86
Q9BVY9
TPECVKGVVTDLLR
39
81-94

IC1_HUMAN (P05155) Plasma
5.14E-03
55119.49
P05155
FQPTLLTLPR
40
391-400

protease C1 inhibitor precursor

(C1 Inh) (C1Inh)

KGP1B_HUMAN (P1461 9)
3.56E-03
77754.71
P14619
PATQQAQKQSASTLQGEPRTK
41
55-75

cGMP-dependent protein

kinase 1, beta isozyme (EC

2.7.1.37) (cGK 1 beta) (cGKI

Q5D862 (Q5D862) Ifapsoriasin
7.33E-03
247926.3
Q5D862
ASHFQSHSSERQRHGSSQVWK
42
2325-2345

Q6ZQY5 (Q6ZQY5)
7.81E-03
119930.9
Q6ZQY5
YENFEDPMGTIDK
43
428-440

Hypothetical protein FLJ46795

RB612_HUMAN (Q8IUD2)
2.83E-03
128007.2
Q8IUD2
LLEMLQSKGLSAKATEEDHER
44
312-332

RAB6 interacting protein 2

(ERG protein 1)

S13A3_HUMAN (Q8WWT9)
1.14E-03
66797.6
Q8WWT9
AQETVPWNIILLLGGGFAMAK
45
421-441

Solute carrier family 13 member

3 (Sodium-dependent high-

affinity dicarboxylat

UBP29_HUMAN (Q9HBJ7)
2.80E-03
104090.1
Q9HBJ7
NMLKEIDKTSFYSICNK
46
124-140

Ubiquitin carboxyl-terminal

hydrolase 29 (EC 3.1.2.15)

(Ubiguitin thiolesteras

VNN3_HUMAN (Q9NY84)
4.12E-04
56054.02
Q9NY84
EEALLLMNKNIDVLEK
47
48-63

Vascular non-inflammatory

molecule 3 precursor (Vanin-3)

MAK_HUMAN (P20794)
3.64E-03
70536.42
P20794
DMKPENLLCMGPELVK
48
125-140

Serine/threonine-protein kinase

MAK (EC 2.7.1.37) (Male germ

cell-associated kin

RPB2_HUMAN (P30876) DNA-
4.42E-03
133810.7
P30876
NKTQISLVR
49
1142-1150

directed RNA polymerase II 140

kDa polypeptide (EC 2.7.7.6)

(RNA polymerase

UBE2I_HUMAN (P63279)
3.13E-03
17995.2
P63279
LRMLFKDDYPSSPPK
50
60-74

Ubiquitin-conjugating enzyme

E2 I (EC 6.3.2.19) (Ubiquitin-

protein ligase I) (

PEBB_HUMAN (Q13951) Core-
3.46E-03
21494.7
Q13951
EYVDLEREAGKVYLK
51
84-98

binding factor, beta subunit

(CBF-beta) (Polyomavirus

enhancer binding pro

Q5NV64 (Q5NV64) V2-11
4.01E-06
10532.11
Q5NV64
DSERPSGIPER
52
50-60

protein (Fragment)

Q6GMV9 (Q6GMV9)
2.04E-08
25629.86
Q6G MV9
YLAWYQQKPGQAPR
53
53-66

Hypothetical protein

Q7Z2U7 (Q7Z2U7)
1.41E-03
24999.29
Q7Z2U7
LTVLRQPK
54
125-132

Hypothetical protein

FOG1_HUMAN (Q8IXO7) Zinc
8.46E-03
104481.6
Q81X07
PTEEEPGSPWSGPDELEPVVQDGQ
55
77-101

finger protein ZFPM1 (Zinc

R

finger protein multitype 1)

(Friend of GATA pro

Q8N4L2 (Q8N4L2) Hypothetical
3.39E-03
28062.26
Q8N4L2
FNTLAKCPHCKK
56
168-179

protein TMEM55A

Q9H8C8 (Q9H8C8)
4.38E-03
148896.6
Q9H8C8
LTEVADFHHAASKVLR
57
188-203

Hypothetical protein FLJ13755

CRP_HUMAN (P02741) C-
9.64E-06
25022.68
P02741
KAFVFPK
58
25-31

reactive protein precursor

[Contains: C-reactive protein(1-

205)]

CRP_HUMAN (P02741) C-
9.64E-06
25022.68
P02741
RQDNEILIFWSK
58
76-87

reactive protein precursor

[Contains: C-reactive protein(1-

205)]

CRP_HUMAN (P02741) C-
9.64E-06
25022.68
P02741
YEVQGEVFTKPQLWP
58
210-224

reactive protein precursor

[Contains: C-reactive protein(1-

205)]

CFAI_HUMAN (P05156)
3.59E-04
65676.66
P05156
HGNTDSEGIVEVK
59
118-130

Complement factor I precursor

(EC 3.4.21.45) (C3B/C4B

inactivator) [Contains: C

DPP4_HUMAN (P27487)
8.28E-03
88222.52
P27487
RIQNYSVMDICDYDESSGRWNCLV
60
318-343

Dipeptidyl peptidase 4 (EC

AR

3.4.14.5) (Dipeptidyl peptidase

IV) (DPP IV) (T-cell

APOF_HUMAN (Q13790)
2.03E-06
33442.05
Q13790
SGVQQLIQYYQDQK
61
233-246

Apolipoprotein F precursor

(Apo-F) (Lipid transfer inhibitor

protein) (LTIP)

PUM1_HUMAN (Q14671)
6.75E-03
126394.8
Q14671
MIDVAEPGQR
62
1128-1137

Pumilio homolog 1 (Pumilio-1)

(HsPUM)

Q5NV67 (Q5NV67) V1-11
2.53E-11
10481.04
Q5NV67
LLIYYDDLLPSGVSDR
63
47-62

protein (Fragment)

Q68CZ6 (Q68CZ6)
9.82E-03
69606.88
Q68CZ6
WAEESLHSLTSK
64
292-303

Hypothetical protein

DKFZp686I1 868

LCTL_HUMAN (Q6UWM7)
4.68E-03
65046.79
Q6UWM7
ELHVNHYRFSLSWPR
65
102-116

Lactase-like protein precursor

(Klotho/Lactase-phlorizin

hydrolase-related prot

TECTB_HUMAN (Q96PL2)
6.77E-03
36931.6
Q96PL2
EAPFVLEASEIGSDLFAGVEAKGL
66
170-198

Beta-tectorin precursor

SIRFK

UHRF1_HUMAN (Q96T88)
5.95E-03
89756.99
Q96T88
LGLTMQYPEGYLEALANRER
67
601-620

Ubiquitin-like containing PHD

and RING finger domains

protein 1 (EC 6.3.2.-) (

TRIM7_HUMAN (Q90029)
7.86E-03
56595.02
Q9C029
KELEDCEVFRSTEK
68
186-199

Tripartite motif protein 7

(Glycogenin-interacting protein)

(RING finger prote

RMP_HUMAN (O94763) RNA
1.85E-03
56729.11
094763
KLLPLSVTPEAFSGTVIEK
69
439-457

polymerase II subunit 5-

mediating protein (RPB5-

mediating protein)

Q5NV69 (Q5NV69) V1-13
6.37E-09
10328.92
Q5NV69
LLIYGNSNRPSGVPDR
70
48-63

protein (Fragment)

Q68DN6 (Q68DN6)
5.28E-03
196563.1
Q68DN6
FDAEISQWKERGLGNLK
71
1054-1070

Hypothetical protein

DKFZp781D1923

Q6XYE5 (Q6XYES) FP17548
7.90E-03
23803.52
Q6XYE5
GTLPETPPLPLRSEMR
72
178-193

Q96MC4 (Q96MC4)
1.89E-036
9795.48
Q96MC4
AELEKER
73
536-542

Hypothetical protein FLJ32655

SRPK1_HUMAN (Q96SB4)
5.28E-03
92349.12
Q96SB4
SAEHYTETALDEIRLLK
74
284-300

Serine/threonine-protein kinase

SRPK1 (EC 2.7.1 .37)

(Serine/arginine-rich prot

Q9H371 (Q9H371) PR01495
6.96E-03
8858.949
Q9H371
SHHPSESLLTVGSILNSLVRLQRS
75
51-74

KINH_HUMAN (P33176)
2.44E-03
109616.9
P33176
VHEMEKEHLNK
76
680-690

Kinesin heavy chain

(Ubiquitous kinesin heavy

chain) (UKHC)

UBE3A_HUMAN (Q05086)
2.30E-03
100581.4
Q05086
AFRRGFHMVTNESPLK
77
732-747

Ubiquitin-protein ligase E3A

(EC 6.3.2.-) (E6AP ubiquitin-

protein ligase) (Onc

QSNV85 (Q5NV85) V2-8
5.35E-07
10508.96
Q5NV85
ITCGGNNIGSK
78
20-30

protein (Fragment)

NOTC4_HUMAN (Q99466)
2.54E-03
209478.2
Q99466
APSCGFHHCHHGGLCLPSPK
79
1088-1107

Neurogenic locus notch

homolog protein 4 precursor

(Notch 4) (hNotch4) [Contai

ZNF71_HUMAN (Q9NQZ8)
8.47E-03
54462.41
Q9NQZ8
AFSQNMHLIVHQRTHTGEK
80
250-268

Endothelial zinc finger protein

induced by tumor necrosis

factor alpha (Zinc f

Q9UL82 (Q9UL82) Myosin-
8.88E-05
11437.69
Q9UL82
DTERPSGIPER
81
50-60

reactive immunoglobulin light

chain variable region

(Fragment)

APOD_HUMAN (P05090)
2.08E-05
21261.77
P05090
IPTTFENGR
82
52-60

Apolipoprotein D precursor

(Apo-D) (ApoD)

APOD HUMAN (P05090)
2.08E-05
21261.77
P05090
VLNQELR
82
76-82

Apolipoprotein D precursor

(Apo-D) (ApoD)

Q5NV81 (Q5NV81) V1-16
3.96E-09
10277.88
QSNV81
LLIYSNNQRPSGVPDR
83
47-62

protein (Fragment)

O95267 (O95267) Calcium and
1.35E-03
90272.02
095267
ATQTESQPWIGSEGPSGPFVLSSP
84
672-696

DAG-regulated guanine

R

nucleotide exchange factor II

LV1D_HUMAN (P01702) Ig
4.33E-04
11446.54
P01702
LLIYDNNKRPSGIPDR
85
47-62

lambda chain V-I region NIG-64

LV2D_HUMAN (P01707) Ig
7.97E-06
11553.64
P01707
SGDTASLTISGLR
86
69-81

lambda chain V-II region TRO

HBAZ_HUMAN (P02008)
2.21E-03
15496.2
P02008
VDPVNFK
87
93-99

Hemoglobin zeta subunit

(Hemoglobin zeta chain) (Zeta-

globin) (HBAZ)

Q15043 (QQ5043) KIAAOO62
1.27E-03
58379.58
Q15043
LLLPPPAAWDLAVRLR
88
10-25

protein (Fragment)

Q5VWOO (Q5VWOO) Novel
4.91E-03
50770.95
Q5VWOO
FLEERASSSLDSMPGPAGR
89
382-400

protein

Q68C18 (Q68C18) Hypothetical
3.80E-03
48598.88
Q68C18
ELDIGISATYCGAHSVPK
90
199-216

protein HMFT1272 (Fragment)

Q6NS96 (Q6NS96) IGLV3-21
5.62E-07
24883.22
Q6NS96
LSGSNSGNTATLTISR
91
80-95

protein

Q6S362 (Q6S362) Usher
6.05E-03
575234.6
Q6S362
LSSATPTSLQVVWSTPAR
92
2443-2460

syndrome 2A isoform B

(Autosomal recessive, mild)

SMS1_HUMAN (Q86VZ5)
3.25E-04
49175
Q86VZ5
EMIKIPMPELERSQYPMEWGK
93
120-140

Phosphatidylcholine:ceramide

cholinephosphotransferase 1

(EC 2.7.-.-) (Transmem

RYR2_HUMAN (Q92736)
3.02E-03
564136.1
Q92736
AFLDNAAEDLEK
94
3059-3070

Ryanodine receptor 2 (Cardiac

muscle-type ryanodine

receptor) (RyR2) (RYR-2) (C

ZMY15_HUMAN (Q9H091)
7.23E-03
77264.05
Q9H091
ELESLVPR
95
277-284

Zinc finger MYND domain

containing protein 15

KV1E_HUMAN (P01597) Ig
1.17E-03
11653.77
P01597
YLNWYQQKPGK
96
32-42

kappa chain V-I region DEE

IGF1B_HUMAN (P05019)
7.48E-03
21826.96
P05019
NAECRGK
97
186-192

Insulin-like growth factor IB

precursor (IGF-IB)

(Somatomedin C) (Mechano

grow

SC24C_HUMAN (P53992)
1.25E-03
118239.2
P53992
GTEPFVTGVRGQVPPLVTTNFLVK
98
341-364

Protein transport protein

Sec24C (SEC24-related protein

C)

CENB2_HUMAN (Q15057)
7.20E-03
87973.21
Q15057
LDKLVKLCIAMIDTGK
99
35-50

Centaurin-beta 2 (Cnt-b2)

Q5JZ85 (Q5JZ85)
4.18E-03
24449.01
Q5JZ85
SYSCQVMHEGSTAEK
100
202-216

OTTHUMP00000028776

Q5NV79 (Q5NV79) V5-4
2.91E-06
10672.02
Q5NV79
FSGSSSGADR
101
66-75

protein (Fragment)

Q5SGD5 (Q5SGD5) Maguin-
3.65E-03
61848.88
Q5SGD5
SSSTEPSLLVSWFTR
102
535-549

like protein variant II (Maguin-

like protein variant III) (Maguin-

like prote

Q8N5F4 (Q8N5F4) IGLC1
2.23E-04
24945.16
Q8N5F4
ITCSGDALPK
103
39-48

protein

LV1E_HUMAN (P01 703) Ig
2.75E-04
10897.52
P01703
VFGGGTK
104
92-98

lambda chain V-I region NEWM

C1QA_HUMAN (P02745)
2.95E-06
26000.19
P02745
KGHIYQGSEADSVFSGFLIFPSA
105
223-245

Complement C1q

subcomponent, A chain

precursor

QSNV82 (QSNV82) V4-2
7.12E-11
11135.42
QSNV82
IYWYQQKPGSPPQYLLR
106
35-51

protein (Fragment)

Q6ZTG8 (Q6ZTG8)
3.99E-03
115755
Q6ZTG8
ELCWLLRDER
107
752-761

Hypothetical protein FLJ44670

Q8TBC9 (Q8TBC9) IGLV3-25
3.35E-04
24851.22
Q8TBC9
DNERPSGIPER
108
69-79

protein

FA38A_HUMAN (Q92508)
9.65E-03
232890.5
Q92508
RPSRSGGR
109
1426-1433

Protein FAM38A

Q96TB2 (Q96TB2) Envelope
5.65E-04
19697.06
Q96TB2
IVCLPSGIFFVCGTSAYR
110
55 - 72

protein (Fragment)

DACT1_HUMAN (Q9NYF0)
2.50E-03
90118.62
Q9NYF0
EAWAKPK
111
663-670

Dapper homolog 1 (hDPR1)

(Heptacellular carcinoma novel

gene 3 protein)

Q9UL86 (Q9UL86) Myosin-
8.61E-04
11920.96
Q9UL86
LLIYGTSSR
112
47-55

reactive immunoglobulin kappa

chain variable region

(Fragment)

AFF3_HUMAN (P51826)
1.57E-04
133651.8
P51826
PKADSQLQPHGGDLTK
113
917-932

AF4/FMR2 family member 3

(LAF-4 protein) (Lymphoid

nuclear protein related to A

Q5T8Z0 (Q5T8Z0)
8.09E-04
18172.26
Q5T8Z0
STLSIGRSLPHITDVSWRLEYQIK
114
81-104

Chromosome 10 open reading

frame 8 (Fragment)

Q8TE63 (Q8TE63)
4.70E-05
11471.7
Q8TE63
LLIYDNNKRPSGVPDR
115
47-62

Immunglobulin light chain

variable region (Fragment)

Q96LP2 (Q96LP2) Hypothetical
6.45E-03
47696.67
Q96LP2
KLSQNIEILEDQIRAR
116
135-150

protein FLJ25333

CK5P1_HUMAN (Q96SZ6)
7.74E-05
67645.74
Q96SZ6
ITSASSQTLR
117
576-585

CDK5 regulatory subunit-

associated protein 1 (CDK5

activator-binding protein C

SNPH_HUMAN (O15079)
1.28E-03
57953.73
O15079
RQGQPIYNISSLLR
118
493-506

Syntaphilin

UNC5C_HUMAN (O95185)
2.45E-03
103036
O95185
RTRTCTNPAPLNGGAFCEGQSVQK
119
283-306

Netrin receptor UNOSO

precursor (Unc-5 homolog C)

(Unc-5 homolog 3)

PRS8_HUMAN (P621 95) 26S
4.74E-03
45597.1
P62195
IEFPPPNEEARLDILK
120
315-330

protease regulatory subunit 8

(Proteasome subunit p45)

(p45/SUG) (Proteasom

OCRL_HUMAN (Q01968)
5.61E-03
104138
Q01968
EPCALTLAQRNGQYELIIQLHEK
121
26-48

Inositol polyphosphate 5-

phosphatase OCRL-1 (EC

3.1.3.36) (Lowe's oculocerebror

M4K2_HUMAN (Q12851)
1.21E-03
91528.26
Q12851
TRWTQNFHHFLK
122
239-250

Mitogen-activated protein

kinase kinase kinase kinase 2

(EC 2.7.1.37) (MAPKIERK

TP53B_HUMAN (Q12888)
8.07E-03
213440.8
Q12888
AADISLDNLVEGKRK
123
1614-1628

Tumor suppressor p53-binding

protein 1 (p53-binding protein

1) (53BP1)

Q5BJF6 (Q5BJF6) ODF
27.71E-03
95341.83
Q5BJF6
KIDSLMNAVGCLKSEVK
124
126-142

protein

Q6PK04 (Q6PKO4) MGC16597
6.85E-03
33211.11
Q6PK04
NQAIRQPEVQAAPK
125
142-155

protein

Q6ZTZ5 (Q6ZTZ5) Hypothetical
8.36E-03
109095.2
Q6ZTZ5
TEVEISDHNDSLLMKPLRFR
126
68-87

protein FLJ44101

Q6ZUU4 (Q6ZUU4)
3.16E-04
65663.22
Q6ZUU4
KMKATNSLGAGIIEK
127
415-429

Hypothetical protein FLJ43328

TB182_HUMAN (Q9C002) 182
2.01E-03
181703.8
Q9COC2
DEDGSTLFRGWSQEGPVK
128
210-227

kDa tankyrase 1-binding protein

GRLF1_HUMAN (Q9NRY4)
8.92E-03
172119.9
Q9NRY4
GDNAVIPYETDEDPRRR
129
1200-1216

Glucocorticoid receptor DNA-

binding factor 1 (Glucocorticoid

receptor repressi

U171_HUMAN (Q12980)
5.46E-03
63564.88
Q12980
RIATVLQHEER
130
148-158

CGTHBA protein (-14 gene

protein)

Q8IXL9 (Q8IXL9) 10 motif
6.96E-03
19614.73
Q8IXL9
RQAALIAYATR
131
89-99

containing F2

PVR2_HUMAN (Q92692)
3.34E-03
57706.01
Q92692
PKNQAEAQK
132
162-170

Poliovirus receptor related

protein 2 precursor (Herpes

virus entry mediator B)

MYO5C_HUMAN (Q9NQX4)
1.73E-03
202665.4
Q9NQX4
LYNNFVNRNPLFEK
133
525-538

Myosin-5C (Myosin Vc)

Q9UHB7 (Q9UHB7) AF5q31
1.25E-04
127381.5
Q9UHB7
AHLTKLK
134
293-299

protein

BAIP2_HUMAN (Q9UQB8)
6.61E-03
60829.68
Q9UQB8
SLSPPQSQSKLSDSYSNTLPVR
135
323-344

Brain-specific angiogenesis

inhibitor 1-associated protein 2

(BAI1-associated

CAD13_HUMAN (P55290)
1.05E-03
78238.09
P55290
QAVPDKVWKISK
136
625-636

Cadherin-1 3 precursor

(Truncated-cadherin) (T-

cadherin) (T-cad) (Heart-

cadheri

NOG2_HUMAN (Q13823)
5.25E-03
83603.43
Q13823
QISVGFIGYPNVGKSSVINTLRSK
137
310-333

Nucleolar GTP-binding protein

2 (Autoantigen NGP-1)

Q6ZN84 (06ZN84)
6.46E-05
65950.16
Q6ZN84
SFLFDKRPLSPALNALK
138
318-334

Hypothetical protein FLJ16339

Q8N549 (Q8N549) Hypothetical
1.17E-03
27915
Q8N549
KFLALQSKNSDADFQNNEK
139
107-125

protein C8orf36

Q8WYY5 (Q8WYY5)
2.14E-03
11844.71
Q8WYY5
KMEHHFLATIGLFFWAQGGK
140
18-38

Hypothetical_protein

CN133 HUMAN (Q9H9C1)
1.47E-03
56970.36
Q9H9C1
VCSLERFRSLQDK
141
169-181

Protein C14orf133

DYH9_HUMAN (Q9NYC9)
2.61E-03
511604.4
Q9NYC9
LANPHYQPELQAQATLINFTVTR
142
3557-3579

Ciliary dynein heavy chain 9

(Axonemal beta dynein heavy

chain 9)

Q9UPW7 (Q9UPW7) KIAA1033
5.53E-03
138341.3
Q9UPW7
FYIFSQFMYDEHIK
143
873-886

protein (Fragment)

DYN3_HUMAN (Q9UQ16)
2.42E-03
96621.48
Q9UQ16
TGLFTPDMAFEAIVKK
144
400-415

Dynamin-3 (EC 3.6.5.5)

(Dynamin, testicular) (T-

dynamin)

HCFC2_HUMAN (Q9Y5Z7)
8.55E-03
86724.45
Q9Y5Z7
ILVFGGMVEYGRYSNELYELQASR
145
84-111

Host cell factor 2 (HCF-2) (C2

WLWK

factor)

O00319 (O00319)
7.88E-03
74438.16
O00319
HFNIHSWEKK
146
512-521

WUGSC:H_DJ525N14.1

protein

PHF2_HUMAN (O75151) PHD
1.57E-03
121156.9
O75151
KRVLNVTNLEFSDTR
147
187-201

finger protein 2 (GROS)

PLST_HUMAN (P13797) T-
4.92E-04
70391.24
P13797
AVGDGIVLCKMINLSVPDTIDER
148
156-178

plastin (Plastin-3)

Q5KSL6 (Q5KSL6)
6.79E-03
141738.7
Q5KSL6
GLYDDTTAFLDEK
149
1154-1166

Diacylglycerol kinase kappa

Q6PJZ1 (Q6PJZ1) C9orf58
2.36E-03
19565.95
Q6PJZ1
RLAEINREFLCDQK
150
51-64

protein (Fragment)

ARP10_HUMAN (Q9NZ32)
2.74E-03
46277.34
Q9NZ32
ANCVAWLGGAIFGALQDILGSRSV
151
350-375

Actin-related protein 10

SK

(hARP11)

MCE1_HUMAN (O60942)
6.87E-04
68512.69
O60942
FHPSMLSNYLKSLKVK
152
45-60

mRNA capping enzyme (HCE)

(HCAP1) [Includes:

Polynucleotide 5-

triphosphatase

STXB2_HUMAN (Q15833)
1.71E-03
66396.59
Q15833
NGVSEENLAK
153
415-424

Syntaxin binding protein 2

(Unc-18 homolog 2) (Unc-18B)

(Unc18-2)

KLH17_HUMAN (Q6TDP4)
7.02E-03
69830.2
Q6TDP4
QVLELVSSDSLNVPSEEEVYR
154
237-257

Keich-like protein 17 (Actinfilin)

ANC4_HUMAN (Q9UJXS)
7.70E-04
91933.59
Q9UJX5
YEPLGLDAAGIEEAITAVGSFILK
155
376-399

Anaphase promoting complex

subunit 4 (APC4) (Cyclosome

subunit 4)

SRPX_HUMAN (P78539) Sushi
6.50E-03
51538.23
P78539
IQYTVYDRAENK
156
237-248

repeat-containing protein SRPX

precursor

ZN11B_HUMAN (Q06732) Zinc
9.02E-04
90624.22
Q06732
AHTGEKSCQCNECGK
157
714-728

finger protein 11B

Q6ZTB6 (Q6ZTB6)
2.55E-03
14350.79
Q6ZTB6
RVGHAGFELLTSGDLPASASQSAG
158
75-105

Hypothetical protein FLJ44808

IAGVSHR

VASHL_HUMAN (Q86V25)
2.71E-03
40424.05
Q86V25
SRSSHARPVSLATSGGSEEEDK
159
22-43

Vasohibin-like protein

CKSP2_HUMAN (Q96SN8)
2.75E-03
214903.6
Q96SN8
KMHEGDLAMALVLDEK
160
204-219

CDK5 regulatory subunit-

associated protein 2 (CDKS

activator-binding protein C

FBX6_HUMAN (Q9NRD1) F-
8.94E-03
33910.88
Q9NRD1
VTNSSIWSPKMTR
161
250-263

box only protein 6 (F-box/G-

domain protein 2)

MGR5_HUMAN (P41594)
6.46E-03
132383.6
P41594
EMGKDYFDYINVGSWDNGELK
162
473-493

Metabotropic glutamate

receptor 5 precursor (mGIuR5)

AP3S2_HUMAN (P59780)
5.33E-04
22003.34
P59780
NINLPEIPRNINIGDLNIKVPNLSQFV
163
167-193

Adapter-related protein

complex 3 sigma 2 subunit

(Sigma-adaptin 3b) (AP-3 com

Q5H9U9 (Q5H9U9)
9.27E-03
152084.4
Q5H9U9
VYRAEYINFLENLK
164
1181-1194

Hypothetical protein

DKFZp781D1175

Q5VWL1 (Q5VWL1)
3.08E-04
162847.8
Q5VWL1
NWEMAYTDTGMIYFIDHNTKTTTW
165
298-325

Membrane-associated

LDPR

guanylate kinase-related 3

(MAGI-3)

Q8WVY7 (Q8WVY7) Ubiquitin-
4.36E-03
36781.22
Q8WVY7
KNTIMFDDIGRNFLMNPQNGLK
166
246-267

like domain containing CTD

phosphatase 1 (CTD-like

phosphatase domain-con

LRP1B_HUMAN (Q9NZR2)
3.60E-03
515158.1
Q9NZR2
NGVFRVQKFGHGSVEYLALNIDK
167
4135-4157

Low-density lipoprotein

receptor-related protein 1B

precursor (Low-density lip

REV1_HUMAN (Q9UBZ9) DNA
1.37E-03
138161.2
Q9UBZ9
DQTKCAASVGIGSNILLAR
168
600-618

repair protein REVi (EC 2.7.7.-

)(Rev1-like terminal

deoxycytidyl transfera

CU005_HUMAN (Q9Y3R5)
4.39E-03
258059.3
Q9Y3R5
KNGGEWDVEKVVIDLGGSR
169
725-743

Protein C21orf5

NFIC_HUMAN (P08651)
3.05E-03
55640.11
P08651
SPFNSPSPQDSPR
170
333-345

Nuclear factor 1 C-type

(Nuclear factor 1/C) (NF1-C)

(NFI-C) (NE-I/C) (CCAAT-bo

ITA3_HUMAN (P26006)
4.27E-03
118622.2
P26006
DKLRPIIISMNYSLPLR
171
563-579

Integrin alpha-3 precursor

(Galactoprotein B3) (GAPB3)

(VLA-3 alpha chain) (CD4

AP2B1_HUMAN (P63010)
4.23E-03
104486.2
P63010
VIAAMTVGKDVSSLFPDVVNCMQT
172
37-67

Adapter-related protein

DNLELKK

complex 2 beta 1 subunit (Beta-

adaptin) (Plasma membra

Q6ZSJ3 (Q6ZSJ3) Hypothetical
1.08E-03
18675.65
Q6ZSJ3
VLLAFRGRGSGTDR
173
124-137

protein FLJ45478

SEC8_HUMAN (Q96A65)
7.30E-03
110428.6
Q96A65
DASVPLIDVTNLPTPR
174
224-239

Exocyst complex component

Sec8

PLCB1_HUMAN (Q9NQ66) 1-
6.38E-03
138479.2
Q9NQ66
AKQLAALTLEDEEEVK
175
824-839

phosphatidylinositol-4,5-

bisphosphate

phosphodiesterase beta 1 (EC

3.1 .4.11)

In addition, the tandem mass spectra were analyzed using more stringent filtering criteria, with a goal of reducing false positives. In particular, the spectra were analyzed using the filtering alorithms of the Scalfold Software (Proteome Software Inc., Portland Oreg.).

The results are provided in Table 2. In short, 74 peptides were identified that correlate to the disease state. Thus, evaluating patient samples for the presence of one or more of these biomarkers will provide a useful method for detecting colorectal cancer.

TABLE 2

Calculated

Protein

SEQ
Peptide

accession
Protein

ID
Mass
Residue

Protein name
numbers
MW (Da)
Peptide sequence
NOs
(AMU)
Number

C-reactive protein precursor
CRP_HUMAN
25021
KAFVFPK
176
836.5
25-31

[Contains: C-reactive

protein(1-205)]

C-reactive protein precursor
CRP_HUMAN
25021
RQDNEILIFWSK
177
1548.8
76-87

[Contains: C-reactive

protein(1-205)]

Ig kappa chain V-I region
KV1J_HUMAN
12751
ASQSISSWLAWYQQKPGK
178
2065.1
47-64

HK102 precursor

(Fragment)

Apolipoprotein D precursor
APOD_HUMAN
21258
IPTTFENGR
179
1034.5
52-60

(Apo-D) (ApoD)

Alpha-1B-glycoprotein
A1BG_HUMAN
54254.4
SWVPHTFESELSDPVELLVAES
180
2471.2
474-495

precursor (Alpha-1-B

glycoprotein)

Alpha-1B-giycoprotein
A1BG_HUMAN
54254.4
TPGAAANLELIFVGPQHAGNYR
181
2296.2
448-469

precursor (Alpha-1-B

glycoprotein)

Hypothetical protein
Q6ZNX5
17656.3
RPSGVSDR
182
873.45
73-80

FLJ26936

Ig lambda chain V-IV region
LV4C_HUMAN
11498.7
SYELTQPPSVSVSPGQTAR
183
2004
1-19

Hil

Carbohydrate
CHST8_HUMAN
48817.7
EPFERLVSAFRDK
184
1593.8
259-271

sulfotransferase 8 (EC

2.8.2.-) (N-

acetylgalactosamine-4-O-

sulfotransferase 1)

(GalNAc-4-O-

sulfotransferase 1)

(GalNAc-4-ST1)

(GalNAc4ST-1)

Hypothetical protein
Q8NEJ1
25005.7
NDQRPSGVPDR
185
1240.6
71-81

Amyloid lambda 6 light
Q96JD0
12276.6
LTVLGQPK
186
855.53
109-116

chain variable region SAR

(Fragment)

C18orf34 protein
Q5BJE1
100177
YESEIKYLTIMKLK
187
1759
477-490

Hypothetical protein
Q7Z2U7
24996.3
LTVLRQPK
188
954.61
125-132

FIGNL1 protein
Q6PIW4
74092.8
YHQPQRASGSSYGGVK
189
1721.8
310-325

(AMU)

Serum amyloid A protein
SAA_HUMAN
13514.5
SEESELGEAFOGAR
190
1550.7
20-33

precursor (SAA) [Contains:

Amyloid protein A (Amyloid

fibril protein AA); Serum

amyloid protein A(2-104);

Serum amyloid protein A(3-

104); Serum amyloid

protein A(2-103); Serum

amyloid protein A(2-102);

Serum amyloid protein A(4-

101

SERPINC1 protein
Q8TCE1
29075.3
ANRPFLVFIR
191
1232.7
231-240

Serum amyloid P-
SAMP_HUMAN
25369.7
AYSLFSYNTQGR
192
1406.7
65-76

component precursor (SAP)

(9.5S alpha-1-glycoprotein)

[Contains: Serum amyloid

P-component(1-203)]

Immunoglobulin lambda-like
IGLL1_HUMAN
22944.9
YAASSYLSLTPEQWR
193
1771.9
173-187

polypeptide 1 precursor

(Immunoglobulin-related

14.1 protein)

(Immunoglobulin omega

polypeptide) (Lambda 5)

(CD179b antigen)

Hemoglobin gamma-1
HBG1_HUMAN
15991.3
VNVEDAGGETLGR
194
1316.6
18-30

subunit (Hemoglobin

gamma-i chain) (Gamma-

1-globin) (Hemoglobin

gamma-A chain) (Hb F

Agamma)

Hypothetical protein
Q8N313
81587.4
QIKDETLQAAVR
195
1371.8
414-425

DKFZp761 P18121

Zinc finger protein 40
ZEP1_HUMAN
297199
QFTKQNGETPGIIAEASK
196
1919
107-124

(Human immunodeficiency

virus type I enhancer-

binding protein 1) (HIV-

EP1) (Major

histocompatibility complex

binding protein 1) (MBP-1)

(Positive regulatory domain

II binding factor 1) (PRD!I-

BF1)

Kininogen-1 precursor
KNG1_HUMAN
71927.5
AATGECTATVGK
197
1165.6
102-113

(Alpha-2-thiol proteinase

inhibitor) [Contains:

Kininogen-1 heavy chain;

Bradykinin (Kallidin I);

Lysyl-bradykinin (Kallidin II);

Kininogen-1 light chain; Low

molecular weight growth

promoting factor]

Kininogen-1 precursor
KNG1_HUMAN
71927.5
TVGSDTFYSFK
198
1251.6
65-75

(Alpha-2-thiol proteinase

inhibitor) [Contains:

Kininogen-1 heavy chain;

Bradykinin (Kallidin I);

Lysyl-bradykinin (Kallidin II);

Kininogen-1 light chain; Low

molecular weight growth

promoting factor]

Kininogen-1 precursor
KNG1_HUMAN
71927.5
YNSQNQSNNQFVLYR
199
1874.9
44-58

(Alpha-2-thiol proteinase

inhibitor) [Contains:

Kininogen-1 heavy chain;

Bradykinin (Kallidin I);

Lysyl-bradykinin (Kallidin II);

Kininogen-1 light chain; Low

molecular weight growth

promoting factor]

Ig kappa chain V-II region
KV2A_HUMAN
12658.6
FSGSGSGTDFTLK
200
1303.6
69-81

Cum

Ig kappa chain V-Il region
KV2A_HUMAN
12658.6
FSGSGSGTDFTLKISR
201
1659.8
69-84

Cum

Netrin receptor UNC5C
UNC5C_HUMAN
103085
VYEMYVTVHR
202
1296.6
564-573

precursor (Unc-5 homolog

C) (Unc-5 homolog 3)

VS-4 protein (Fragment)
Q5NV79
10660.8
FSGSSSGADR
203
970.42
66-75

Hypothetical protein
Q9NVY1
45979
WGMLFLVNQLFKIYIK
204
2013.1
166-181

FLJ10440

Hypothetical protein
Q6N095
52341.3
VTVSSASTK
205
879.48
141-149

DKFZp686K03196

Ig lambda chain V-II region
LV2D_HUMAN
11542.9
SGDTASLTISGLR
206
1277.7
69-81

TRO

CCDC28B protein (Novel
Q8TBV8
22018.9
RSPKPCLAQPAQAPGTLR
207
1948.1
8-25

protein)

Oxysterol binding protein-
OSR2_HUMAN
55184.3
FWGKSVEAEPRGTITLELLK
208
2274.3
206-225

related protein 2 (OSBP-

related protein 2) (ORP-2)

Apolipoprotein M (Apo-M)
APOM_HUMAN
21235.9
AFLLTPR
209
817.49
172-178

(ApoM) (G3a protein)

Apolipoprotein M (Apo-M)
APOM_HUMAN
21235.9
KWIYHLTEGSTDLR
210
1718.9
99-112

(ApoM) (G3a protein)

V1-16 protein (Fragment)
Q5NV81
10266.2
LLIYSNNQRPSGVPDR
211
1829
47-62

Ig kappa chain V-I region
KV1A_HUMAN
11974.8
TFGQGTK
212
738.38
97-103

AG

Hypothetical protein
Q6GMV9
25628.4
YLAWYQQKPGQAPR
213
1705.9
53-66

Apolipoprotein F precursor
APOF_HUMAN
33445.8
SGVQQLIQYYQDQK
214
1697.9
233-246

(Apo-F) (Lipid transfer

inhibitor protein) (LTIP)

Hypothetical protein
Q5U5X8
46774
AAVSSSSTAAPAGPAKSVLKSAEGK
215
2272.2
108-132

FLJ14721

V2-8 protein (Fragment)
Q5NV85
10497.3
ITCGGNNIGSK
216
1120.5
20-30

Protocadherin gamma C3
PCDGK_HUMAN
101061
ETGEMFVNDRLDR
217
1581.7
82-94

precursor (PCDH-gamma-

C3) (Protocadherin 43) (PC

43)(Protocadherin 2)

Protein C21orf5
CUOO5_HUMAN
258213
KNGGEWDVEKVVIDLGGSR
218
2058.1
725-743

V2-11 protein (Fragment)
QSNV64
10520.9
DSERPSGIPER
219
1242.6
50-60

Microtubule-actin
MACF1_HUMAN
620397
ALIAEHQTFMEEMTRKQPDVDR
220
2645.3
4960-4981

crosslinking factor 1,

isoforms 1/2/3/5 (Actin

cross-linking family protein

7)(Macrophin-1)

(Trabeculin-alpha) (620 kDa

actin-binding protein)

(ABP620)

Neuronal acetylcholine
ACHAS_HUMAN
53037.9
ALRLLLLVQLVAGRCGLAGAAGGAQR
221
2604.5
10-35

receptor protein, alpha-5

subunit precursor

FLJ35834 protein
Q49AL6
92565.1
HKVTGVEYHNAGTQTVPK
222
1966
258-275

OTTHUMP00000028561
Q5THZ5
196693
PEVAAKPALPTQKPAGTLPR
223
2042.2
176-195

V4-2 protein (Fragment)
Q5NV82
11124.7
IYWYQQKPGSPPQYLLR
224
2137.1
35-51

PHD finger protein 2
PHF2_HUMAN
121218
KRVLNVTNLEFSDTR
225
1792
187-201

(GRC5)

Macrophage-stimulating
RON_HUMAN
152208
HIDPFDLTHFLAQGRR
226
1923
1290-1305

protein receptor precursor

(EC 2.7.1.112) (MSP

receptor) (p185-Ron)

(CDW136) (CD136 antigen)

[Contains: Macrophage-

stimulating protein receptor

alpha chain; Macrophage-

stimulating protein receptor

beta chain]

IBR domain containing
IBRD1_HUMAN
31963.7
TRNQTQHLAPQPVLLSDMLYCLK
227
2726.4
241-263

protein 1

Hypothetical protein
Q8N825
66152.4
PGPCWPGPSSHANGDPVAVAK
228
2101
564-584

FLJ40112

Methylenetetrahydrofolate
Q5JYA3
12407.6
AVIELLEKSGVNLDGKK
229
1813
96-112

dehydrogenase (NADP +

dependent) 1-like

(Fragment)

NOD16
Q7RTR4
119414
TMLPEATLLIMIR
230
1501.8
323-335

MGC10701 protein
Q9BSY8
5306.9
MAVTGAGSGAR
231
977.48
1-11

Zinc finger protein 462
ZN462_HUMAN
161512
SLLENAEAK
232
974.52
1402-1410

C14orf125 protein
Q6PSR9
12885.8
QKTLVEQLLSLLNSSPGPPTRK
233
2406.4
49-70

IGLV3-21 protein
Q6N596
24879.8
LSGSNSGNTATLTISR
234
1578.8
80-95

Hypothetical protein
Q6AHZ1
166734
ITSVFSLQSQQASEFLPPEVNQLLQD
235
3258.7
764-792

DKFZp78102147

VLK

Hypotheticalprotein
Q9NUY8
76459.1
VISFIENTSTPVDRHVSSSDRVGK
236
2630.4
505-528

FLJ11046

Zinc finger MYND domain
ZMY15_HUMAN
77295.3
ELESLVPR
237
942.53
277-284

containing protein 15

Band 4.1-like protein 2
E41L2_HUMAN
112570
LVSPEQPPKAK
238
1193.7
497-507

(Generally expressed

protein 4.1) (4.1G)

IGLC1 protein
Q8NSF4
24942.4
ITCSGDALPK
239
1061.5
39-48

Transcription factor BTF3
BT3L1_HUMAN
12518.3
QAEVHTGR
240
897.45
14-21

homolog 1 (Basic

transcription factor 3-like 1)

Complement factor I
CFAI_HUMAN
65701.8
HGNTDSEGIVEVK
241
1384.7
118-130

precursor (EC 3.4.21.45)

(C3B/C4B inactivator)

[Contains: Complement

factor I heavy chain;

Complement factor I light

chain]

V1-11 protein (Fragment)
Q5NV67
10469.7
LLIYYDDLLPSGVSDR
242
1839
47-62

Splice variant of four and a
Q5TM15
15872.1
NNMPCSAFSAKSPSPR
243
1750.8
98-113

half LIM domain protein 2

RNA polymerase II subunit
RMP_HUMAN
56746.4
KLLPLSVTPEAFSGTVIEK
244
2029.2
439-457

5-mediating protein (RPB5-

mediating protein)

T-complex protein 1, theta
TCPQ_HUMAN
59471.8
LYAVHQEGNKNVGLDIEAEVPAVK
245
2593.4
466-489

subunit (TCP-1 -theta)

(CCT-theta)

Seven transmembrane helix
Q8NH06
33619.8
EDTAAAVMYTVVTPLLNPFIYSMR
246
2702.4
258-281

receptor

Lysozyme C precursor (EC
LYSC_HUMAN
16518.9
STDYGIFQINSR
247
1400.7
69-80

3.2.1.17) (1,4-beta-N-

acetylmuramidase C)

WAP four-disulfide core
Q5JYQ5
17950.6
RSLNNLQDLAHNPGLSPR
248
2002.1
141-158

domain 6

PML-RARA regulated
Q8N6W7
73980.3
EKDPQPQQLPPMDPKLLK
249
2102.1
547-564

adaptor molecule 1

Example 2
Identification of Biomarkers for Colorectal Cancer Using LTQ-Orbitrap

Collection of blood and harvest of LMW protein were performed as described in Example 1.

Sample Preparation for Mass Spectrometric Analysis

LMW proteins collected from PrepCell were concentrated by Centricon (Millipore), loaded to SDS-PAGE (4-20% Tris-Glycine, Invitrogen) and proteins were separated by electrophoresis. After Coomassie staining and destaining of the gel, each lane was sliced to 5 bands. Then in-gel digestion by trypsin was performed for each band and peptides were extracted from the gel for mass spectrometric analysis.

Nanoflow Reversed-Phase Liquid Chromatography Tandem Mass Spectrometry

The peptides from each band were analyzed by reversed-phase liquid chromatography nanospray tandem mass spectrometry using LTQ-Orbitrap mass spectrometer (ThermoFisher). Reverse phase column was slurry-packed in-house with 5 μm, 200 Å pore size C₁₈resin (Michrom BioResources, CA) in 100 μm i.d.×10 cm long fused silica capillary (Polymicro Technologies, Phoenix, Ariz.) with a laser-pulled tip. After sample injection, the column was washed for 5 min with mobile phase A (0.1% formic acid) and peptides were eluted using a linear gradient of 0% mobile phase B (0.1% formic acid, 80% acetonitrile) to 50% mobile phase B in 90 min at 200 nl/min, then to 100% B in an additional 5 min. The LTQ-Orbitrap mass spectrometer was operated in a data-dependent mode in which each full MS scan was followed by five MS/MS scans where the five most abundant molecular ions were dynamically selected and fragmented by collision-induced dissociation (CID) using a normalized collision energy of 35%.

Bioinformatic Analysis

Tandem mass spectra were matched against human database downloaded from the National Center for Biotechnology Information (NCBI) through the Sequest Bioworks Browser (ThermoFisher) using full tryptic cleavage constraints and static cysteine alkylation by iodoacetamide. For a peptide to be considered legitimately identified, it had to be the top number one matched and had to achieve cross correlation scores of 1.9 for [M+H]¹⁺, 2.2 for [M+2H]²⁺, 3.5 for [M+3H]³⁺, ΔCn>0.1, and a maximum probabilities of identification of 0.01.

The results are provided in Table 3. In summary, 139 peptides were identified that correlate to the disease state.

Subsequently, the candidate biomarkers are verified and validated for colorectal cancer, followed by analysis of LMW protein fractions less than 25 KDa and less than 15 KDa from colorectal cancer pooled sera by reverse phase protein array.

LMW protein fractions from individual patient samples with and without colorectal cancer were isolated and collected using continuous denaturing electrphoresis and spotted on a nitrocellulose substratum using a reverse phase array format whereby the LMW sample is diluted 1:1 with SDS sample buffer and printed. The slide is then blocked with casein hydrolysate and incubated with an rabbit polyclonal anti-CRP antibody for 16 hours. The slide is washed and incubated with a horseradish peroxidae coupled goat anti-rabbit and subject to tyrmaide amplification using a colorimetric (DAB) precipitant.

Thus, evaluating patient samples for the presence of one or more of these biomarkers will provide a useful method for detecting colorectal cancer.

TABLE 3

Calculated

Protein

SEQ
Peptide

accession
Protein

ID
Mass
Residue

Protein name
numbers
MW (Da)
Peptide sequence
NOs
(AMU)
Number

serum amyloid A1
gi|40316912|ref|NP_000322.2|
13514.5
FFGHGAEDSLADQAANEWGR
250
2178
86-105

isoform 1 [Homo

sapiens]

serum amyloid A1
gi|4O316912|ref|NP_000322.2|
13514.5
GPGGAWAAEVISDAR
251
1456.7
66-80

isoform 1 [Homo

sapiens]

serum amyloid A1
gi|40316912|ref|NP_000322.2|
13514.5
RGPGGAWAAEVISDAR
252
1612.8
65-80

isoform 1 [Homo

sapiens]

serum amyloid A1
gi|40316912|ref|NP_000322.2|
13514.5
SFFSFLGEAFDGAR
253
1550.7
20-33

isoform 1 [Homo

sapiens]

A-gamma globing
gi|28302131|ref|NP_000550.2|
16110.4
LLVVYPWTQR
254
1274.7
32-41

[Homo sapiens]

delta globin [Homo
gi|4504351|ref|NP_000510.1|
16037.1
WAGVANALAHK
255
1149.7
134-145

sapiens]

serum amyloid A4,
gi|10835095|ref|NP_006503.1|
14789.3
VYLQGLIDYYLFGNSSTVLED
256
2624.3
81-103

constitutive [Homo

SK

sapiens]

apolipoprotein C-III
gi|4557323|ref|NP_000031.1|
10834.3
DALSSVQESQVAQQAR
257
1716.9
45-60

precursor [Homo

sapiens]

apolipoprotein C-III
gi|4557323|ref|NP_000031.1|
10834.3
GWVTDGFSSLK
258
1196.6
61-71

precursor [Homo

sapiens]

platelet factor 4
gi|4505733|ref|NP_002610.1|
10827.5
AGPHCPTAQLIATLK
259
1577.8
63-77

(chemokine (C-X-

C motif) ligand 4)

[Homo sapiens]

platelet factor 4
gi|4505733|ref|NP_002610.1|
10827.5
HITSLEVIK
260
1039.6
54-62

(chemokine (C-X-

C motif) ligand 4)

[Homo sapiens]

platelet factor 4
gi|4505733|ref|NP_002610.1|
10827.5
ICLDLQAPLYK
261
1333.7
82-92

(chemokine (C-X-

C motif) ligand 4)

[Homo sapiens]

platelet factor 4
gi|4505733|ref|NP_002610.1|
10827.5
KICLDLQAPLYK
262
1461.8
81-92

(chemokine (C-X-

C motif) ligand 4)

[Homo sapiens]

apolipoprotein C-
gi|4502161|ref|NP_001637.1|
14535.5
AWFLESK
263
880.46
99-105

IV [Homo sapiens]

apolipoprotein C-
gi|4502161|ref|NP_001637.1|
14535.5
DGWQWFWSPSTFR
264
1699.8
67-79

IV [Homo sapiens]

apolipoprotein C-
gi|4502161|ref|NP_001637.1|
14535.5
ELLETWNR
265
1072.6
56-64

IV [Homo sapiens]

apolipoprotein C-
gi|4502161|ref|NP_001637.1|
14535.5
GFMQTYYDDHLR
266
1545.7
80-91

IV [Homo sapiens]

proteoglycan 4
gi|67190163|ref|NP_005798.2|
151062.3
DQYYNIDVPSR
267
1369.6
1373-1383

[Homo sapiens]

proteoglycan 4
gi|67190163|ref|NP_005798.2|
151062.3
GLPNVVTSAISLPNIR
268
1651
1345-1360

[Homo sapiens]

apolipoprotein E
gi|4557325|ref|NP_000032.1|
36135.5
SWFEPLVEDMQR
269
1536.7
281-292

precursor [Homo

sapiens]

alpha-2-HS-
gi|4502005|ref|NP_001613.1|
39305.4
HTFMGWSLGSPSGEVSHPR
270
2081
318-337

glycoprotein

[Homo sapiens]

serpin peptidase
gi|50659080|ref|NP_001076.2|
47635
AVLDVFEEGTEASAATAVK
271
1908
361-379

inhibitor, clade

A, member 3

precursor [Homo

sapiens]

serpin peptidase
gi|50659080|ref|NP_001076.2|
47635
FNRPFLMIIVPTDTQNIFFMS
272
2659.4
395-416

inhibitor, clade

K

A, member 3

precursor [Homo

sapiens]

serpin peptidase
gi|50659080|ref|NP_001076.2|
47635
ITLLSALVETR
273
1215.7
380-390

inhibitor, clade

A, member 3

precursor [Homo

sapiens]

complement
gi|56786155|ref|NP_758957.2|
25756
FNAVLTNPQGDYDTSTGK
274
1927.9
140-157

component 1, q

subcomponent,

gamma

polypeptide

[Homo sapiens]

coagulation factor
gi|4503625|ref|NP_000495.1|
54714.1
ACIPTGPYPCGK
275
1320.6
163-174

X preproprotein

[Homo sapiens]

coagulation factor
gi|4503625|ref|NP_000495.1|
54714.1
NCELFTR
276
939.44
120-126

X preproprotein

[Homo sapiens]

complement
gi|50345296|ref|NP_00100202
192734.8
AEMADQAAAWLTR
277
1433.7
1279-1291

component 4B
9.1|

preproprotein

[Homo sapiens]

proline rich 11
gi|88703045|ref|NP_060774.2|
40068.3
DTIFPSR
278
835.43
100-106

[Homo sapiens]

angiotensinogen
gi|4557287|ref|NP_000020.1|
53136.8
ALQDQLVLVAAK
279
1268.8
83-94

preproprotein

[Homo sapiens]

complement factor
gi|11321587|ref|NP_002104.1|
37643.3
EIMENYNIALR
280
1365.7
271-281

H-related 1 [Homo

sapiens]

ribonuclease,
gi|4506557|ref|NP_002928.1|
16822.4
ENTEIHEDIWNIR
281
1704.8
70-82

RNase A family, 4

precursor [Homo

sapiens]

ribonuclease,
gi|4506557|ref|NP_002928.1|
16822.4
YCNLMMQR
282
1115.5
52-59

RNase A family, 4

precursor [Homo

sapiens]

serine. (or
gi|39725934|ref|NP_002606.3|
46296.3
DTDTGALLFIGK
283
1250.7
400-411

cysteine)

proteinase

inhibitor, clade F

(alpha-2

antiplasmin,

pigment epithelium

derived factor),

member 1 [Homo

sapiens]

plasminogen-
gi|4505883|ref|NP_002656.1|
10952.8
AFQYHSK
284
880.43
63-69

related protein B2

[Homo sapiens]

plasminogen-
gi|4505883|ref|NP_002656.1|
10952.8
KSSIIIR
285
816.53
81-87

related protein B2

[Homo sapiens]

chemokine (C-C
gi|14589959|ref|NP_116738.1|
12279.6
PGIVFITK
286
874.54
78-85

motif) ligand 14

isoform 2

precursor [Homo

sapiens]

sperm
gi|8394343|ref|NP_059121.1|
17388.5
FYNNHAFEEQEPPEK
287
1878.8
68-82

autoantigenic

protein 17 [Homo

sapiens]

myosin light chain
gi|47132569|ref|NP_444257.2|
161732.9
FRKQIQESEHMK
288
1560.8
1274-1285

kinase isoform 4

[Homo sapiens]

UDP-glucose
gi|9910280|ref|NP_064505.1|
177176.9
EPVYLSGYGVELAIK
289
1637.9
237-251

ceramide

glucosyltransfer-

ase- like 1

isoform 1

[Homo sapiens]

hypothetical
gi|34787409|ref|NP_065069.1|
166783.6
EPANSR
290
673.33
1125-1130

protein LOC57148

[Homo sapiens]

serine (or
gi|50363219|ref|NP_00100223
46719.9
FNKPFVFLMIEQNTK
291
1856
390-404

cysteine)
6.1|

proteinase

inhibitor, clade A

(alpha-a

antiproteinase

antitrypsin),

member 1 [Homo

sapiens]

serine (or
gi|50363219|ref|NP_00100223
46719.9
GTEAAGAMFLEAIPMSIPPEV
292
2259.1
368-389

cysteine)
6.1|

K

proteinase

inhibitor, clade A

(alpha-1

antiproteinase,

antitrypsin),

member 1 [Homo

sapiens]

serine (or
gi|50363219|ref|NP_00100223
46719.9
LSITGTYDLK
293
1110.6
315-324

cysteine)
6.1|

proteinase

inhibitor, clade A

(alpha-1

antiproteinase,

antitrypsin),

member 1 [Homo

sapiens]

serine (or
gi|50363219|re|INP_00100223
46719.9
SPLFMGK
294
779.41
405-411

cysteine)
6.1|

proteinase

inhibitor, clade A

(alpha-1

antiproteinase,

antitrypsin),

member 1 [Homo

sapiens]

serine (or
gi|50363219|ref|NP_00100223
46719.9
SVLGQLGITK
295
1015.6
325-334

cysteine)
6.1|

proteinase

inhibitor, clade A

(alpha-1

antiproteinase,

antitrypsin),

member 1 [Homo

sapiens]

serine (or
gi|50363219|ref|NP_00100223
46719.9
VFSNGADLSGVTEEAPLK
296
1833.9
335-352

cysteine)
6.1|

proteinase

inhibitor, clade A

(alpha-1

antiproteinase,

antitrypsin),

member 1 [Homo

sapiens]

secretoglobin,
gi|50363226|ref|NP_443095.2|
10082.3
LLLSSLGIPVNHLIEGSQK
297
2018.2
59-77

family 3A, member

1 [Homo sapiens]

fibroblast
gi|169335401ref1NP_004451.2|
87697.2
TOQEIKILEENKELENALK
298
2257.2
481-499

activation

protein,

alpha subunit

[Homo sapiens]

PREDICTED:
gi|88979556|ref|XP_945982.|
159016.4
WNHCFLCKKCVK
299
1679.8
408-419

similar to Zinc

finger CCHC

domain-containing

protein 4 isoform

5 [Homo sapiens]

lipopolysaccha-
gi|31652249|ref|NP_004130.2|
53368
LAEGFPLPLLK
300
1197.7
444-454

ride- binding

protein

precursor [Homo

sapiens]

sex hormone-
gi|7382460|ref|NP_001031.2|
43762.6
IALGGLLFPASNLR
301
1441.9
170-183

binding globulin

[Homo sapiens]

platelet factor 4
gi|4505735|ref|NP_002611.1|
11535.1
AEAEEDGDLQCLCVK
302
1736.7
34-48

variant 1 [Homo

sapiens]

peptidyl arginine
gi|6912576|ref|NP_036519.1|
74078
AEAFFPNMVNMLVLGK
303
1812.9
579-594

deiminase, type IV

[Homo sapiens]

discs large
gi|51339031|ref|NP_00100380
75244.3
SLESSQRQEAR
304
1290.6
630-640

homolog-
9.1|

associated protein

1 isoform beta

[Homo sapiens]

caspase 14
gi|6912286|ref|NP_036246.1|
27661.8
GSCFIQTLVDVFTK
305
1614.8
184-197

precursor [Homo

sapiens]

alpha-2-plasmin
gi|11386143|ref|NP_000925.1|
54578.6
LGNQEPGGQTALK
306
1312.7
52-64

inhibitor [Homo

sapiens]

prolactin-induced
gi|4505821|ref|NP_002643.1|
16555.1
FYTIEILKVE
307
1254.7
137-146

protein [Homo

sapiens]

PREDICTED:
gi|113420925|ref|XP_0011309
34923.4
ACEGAEVREAAR
308
1318.6
196-207

hypothetical 79.11

protein [Homo

sapiens]

pericentriolar
gi|34878902|ref|NP_006188.2|
228284.4
LPEMEPLVPRVKEVK
309
1780
1906-1920

material 1 [Homo

sapiens]

CIpX caseinolytic
gi|7242140|ref|NP_006651.2|
69206.7
SASEGSSKKSGSGNSGK
310
1554.7
82-98

protease X

homolog [Homo

sapiens]

leucine rich repeat
gi|42544231|ref|NP_006329.2|
78842.3
RYRVYPEGTLELR
311
1651.9
473-485

neuronal 5

precursor [Homo

sapiens]

5-azacytidine
gi|57863291|ref|NP_00100981
117291.9
CSELKGQLGEAEGENLR
312
1889.9
912-928

induced 1 soform
1.1|

b [Homo sapiens]

profilin family,
gi|40786418|ref|NP_955378.1|
14301.7
CVRADEYSLYAKNENTGVVVV
313
2514.3
73-94

member 4 [Homo

K

sapiens]

PREDICTED:
gi|113427383|ref|XP_0011262
27008.5
HMKIKSLEEMYVFSLPMK
314
2259.1
12-29

similarto
32.1|

ribosomal protein

S2 [Homo sapiens]

small inducible
gi|4506831|ref|NP_002979.1|
9830.9
FIVDYSETSPQCPK
315
1670.8
43-56

cytokine A18

precursor [Homo

sapiens]

chemokine (C-X-C
gi|76563933|ref|NP_00102905
13687.7
ILNTPNCALQIVAR
316
1582.9
49-62

motif) ligand 12
8.1|

(stromal cell-

derived factor 1)

isoform gamma

(Homo sapiens]

heparanase
gi|94721347|ref|NP_006656.21
61134.1
TDFLIFDPK
317
1095.6
99-107

[Homo sapiens]

keratin associated
gi|38490587|ref|NP_941972.1|
25086
PSSSVSLLCRPVCRPAR
318
1942
191-207

protein 10-12

[Homo sapiens]

retinitis
gi|5454016|ref|NP_006260.1|
240645.3
KVQPVDLDKAR
319
1268.7
111-121

pigmentosa RP1

protein [Homo

sapiens]

zinc finger protein
gi|41281441|ref|NP_055514.21
200730.2
ENSRTETTMSPPR
320
1521.7
603-615

646 [Homo

sapiens]

PREDICTED:
gi|113429344|ref|XP_372900.4|
13053.9
SHSAHFFEFLTK
321
1450.7
76-87

similar to D-

dopachrome

decarboxylase (D-

dopachrome

tautomerase)

(Phenylpyruvate

tautomerase II)

[Homo sapiens]

fibulin 1 isoform
gi|34734066|ref|NP_006477.2|
77221.7
AITPPHPASQANIIFDITEGN
322
2475.3
620-642

D [Homo sapiens]

LR

macrophage
gi|4505185|ref|NP_002406.1|
12458.5
LLCGLLAER
323
1044.6
79-87

migration inhibitory

factor

(glycosylation-

inhibiting factor)

[Homo sapiens]

thioredoxin [Homo
gi|50592994|ref|NP_003320.2|
11719.3
TAFQEALDAAGDK
324
1336.6
9-21

sapiens]

PREDICTED:
gi|89037862|ref|XP_945963.1|
155161.2
QCGLHDGLQDTSQDK
325
1701.8
1246-1260

similar to

retrotransposon-

like 1 [Homo

sapiens]

|uiescin Q6
gi|13325075|ref|NP_002817.2|
82560.7
SALYSPSDPLTLLQADTVR
326
2047.1
33-51

isoform a [Homo

sapiens]

pancreas-enriched
gi|19923455|ref|NP_057425.2|
258827.4
VHFEDLVFLR
327
1274.7
1928-1937

phospholipase C

[Homo sapiens]

leader-binding
gi|38045903|ref|NP_937825.1|
49166
SVIMWFAEDK
328
1253.6
113-123

protein 32 isoform

2 [Homo sapiens]

transducer of
gi|56118245|ref|NP_073606.2|
66969.4
EENLLNVPKPLPK
329
1490.9
214-226

regulated CREB

protein 3 [Homo

sapiens]

G-gammaglobin
gi|6715607|ref|NP_000175.1|
16108.5
KVLTSLGDAIK
330
1144.7
67-77

[Homo sapiens]

golgi autoantigen,
gi|4758454|ref|NP_004478.1|
376057.9
EISNLNQLIEEFK
331
1576.8
717-729

golgin subfamily b,

macrogolgin (with

transmembrane

signal), 1 [Homo

sapiens]

PREDICTED:
gi|89031353|ref|XP_934040.1|
25721.4
MNQLILTTGGRINTFHGTKK
332
2230.2
1-20

similar to

eukaryotic

translation

initiation factor 5A

[Homo sapiens]

microtubule-
gi|10346135|ref|NP_055083.1|
37013.6
FQDNLDFIQWFK
333
1600.8
144-155

associated protein,

RP/EB family,

member 2 [Homo

sapiens]

PREDICTED:
gi|113422814|ref|XP_0011325
166806.4
HLGLYESSMMPPGRPR
334
1827.9
118-133

hypothetical
91.1|

protein [Homo

sapiens]

chromosome 9
gi||11641247|ref|NP_071738.1|
17200.2
ASASDGSSFVVAR
335
1253.6
120-132

open reading

framal9 [Homo

sapiens]

pancreatic
gi|38201682|ref|NP_937875.1|
17625.8
SNSSMHITDCR
336
1307.5
103-113

ribonuclease

precursor [Homo

sapiens]

ADP-ribosylation
gi|4502209|ref|NP_001653.1|
20512.1
DAVLLVFANK
337
1089.6
118-127

factor 5 [Homo

sapiens]

protein S (alpha)
gi|4506117|ref|NP_000304.1|
75055.3
SFQTGLFTAAR
338
1198.6
91-101

[Homo sapiens]

fibronectin 1
gi|47132551|ref|NP_997640.1|
266182.2
TYHVGEQWQK
339
1275.6
2336-2345

isoform 2

preproprotein

[Homo sapiens]

myoglobin [Homo
gi|4885477|ref|NP_005359.1|
17166.4
HGATVLTALGGILK
340
1350.8
65-78

sapiens]

hypothetical
gi|56711320|ref|NP_00100187
84154.6
LGDNEETQVR
341
1160.6
330-339

protein
2.2|

LOC145407

[Homo sapiens]

G protein-coupled
gi|89257346|ref|NP_005284.2|
38631.1
PGLLHQGRQR
342
1161.7
226-235

receptor 20 [Homo

sapiens]

lysosomal acid
gi|4557010|ref|NP_001601.1|
48326.8
YEQLQNETR
343
1180.6
162-170

phosphatase 2

precursor [Homo

sapiens]

keratin 24 [Homo
gi|9506669|ref|NP_061889.1|
55128.6
|QSVEADINGLR
344
1201.6
242-252

sapiens]

neuroblastoma-
gi|41393547|ref|NP_056993.2|
268559
FRMTQLTVEK
345
1268.7
832-841

amplified protein

[Homo sapiens]

PREDICTED:
gi|89036937|ref|XP_944641.1|
19303.4
EGEMEVAMQK
346
1183.5
62-71

similar to CDC42-

binding protein

kinase beta [Homo

sapiens]

ubi|uinol-
gi|46593007|ref|NP_003356.2|
52627.9
MAASWCRAATAGAQVLLR
347
1945
1-19

cytochrome c

reductase core

protein I [Homo

sapiens]

syntaxin 8 [Homo
gi|4759188|ref|NP_004844.1|
26889.6
QNLLDDLVTR
348
1186.6
80-89

sapiens]

phospholipase D1,
gi|4505873|ref|NP_002653.1|
124170.4
IAADMSNIIENLDTR
349
1691.8
17-31

phophatidylcholine-

specific [Homo

sapiens]

titin isoform N2-B
gi|20143914|ref|NP_003310.2|
2992949
LSWKMPDDDGGDRIK
350
1748.8
9433-9447

(Homo sapiens]

hypothetical
gi|8923251|ref|NP_060208.1|
125270
TAELSVKEYKEVNEK
351
1766.9
683-697

protein LOC54875

[Homo sapiens]

guanine nucleotide
gi|38788319|ref|NP_005266.2|
68642.7
DSREEVER
352
1019.5
88-95

binding protein-like

1 [Homo sapiens]

hypothetical
gi|75832029|ref|NP_065880.1|
150366.6
SQSADPFLNLEMDAGISNIOR
353
2306.1
1070-1090

protein LOC57589

[Homo sapiens]

pyrin and HIN
gi|41282051|ref|NP_945148.1|
50974.1
LMSEMHSFIQIQK
354
1623.8
379-391

domain family,

member 1 beta 2

isoform [Homo

sapiens]

transportin 1
gi|23510381|ref|NP_694858.1|
42709.1
DVIVR
355
601.37
299-303

[Homo sapiens],
gi|46409366|ref|NP_997255.1|

dedicator of
gi|44889960|ref|NP_982272.1|
230806.7
KEQTHWRQANEK
356
1554.8
1332-1343

cytokinesis 8

[Homo sapiens]

kinesin family
gi|9910266|ref|NP_064627.1|
160145.9
QKETAKCEQQMAK
357
1579.8
943-955

member 15 [Homo

sapiens]

zinc finger protein
gi|24429588|ref|NP_060651.2|
141677.7
KEHGKQMK
358
1001.5
1046-1053

532 [Homo

sapiens]

dehydrogenase E1
gi|38788380|ref|NP_061176.3|
103026.3
IGGSVHLIVNNQLGYTTPAER
359
2239.2
356-376

and transketolase

domain containing

protein 1 [Homo

sapiens]

kinesin family
gi|46852174|ref|NP_008985.3|
80025.3
NKARINEDPK
360
1184.6
347-356

member 3A [Homo

sapiens]

major
gi|4504415|ref|NP_002118.1|
38205.8
THVTHHPVFDYEATLR
361
1923
211-226

histocompatibility

complex, class I, G

precursor [Homo

sapiens]

serine peptidase
gi|74027261|ref|NP_006837.2|
120697.7
ETCDEFRRLLQNGK
362
1765.9
630-643

inhibitor, Kazal

type 5 precursor

[Homo sapiens]

centaurin, gamma
gi|16799069|ref|NP_114152.2|
95027
VDSIGSGRAIPIK
363
1312.8
358-370

3 [Homo sapiens]

PREDICTED:
gi|113425692|ref1XP_0011315
6923.8
HGYIGDLKPLMITELR
364
1872
44-59

similarto
13.1|

ribosomal protein

SiSa [Homo

sapiens]

RAB26, member
gi|46361978|ref|NP_055168.2|
27882.5
TPKSKGASTPAASTLPTANGA
365
2084.1
6-27

RAS oncogene

R

family [Homo

sapiens]

PREDICTED:
gi|89044999|ref|XP_934461.1|
28637.1
LLEMQPKEAPEKDPEQLPK
366
2220.2
96-114

similar to Sorting

nexin-19 [Homo

sapiens]

hypothetical
gi|40786410|ref|NP_955373.1|
64161
GSQLEDQALR
367
1116.6
529-538

protein

LOC374920

[Homo sapiens]

hypothetical
gi|71067125|ref|NP_078857.4|
129514.6
MDYSQEMHLKLSKK
368
1753.9
1-14

protein LOC79632

[Homo sapiens]

fibronectin type III
gi|34222186|ref|NP_660278.2|
38350.5
AVSVNDEDLLVRILQGGR
369
1954.1
117-134

and ankyrin repeat

domains 1 [Homo

sapiens]

PREDICTED:
gi|89035129|ref|XP_945446.1|
20879
EEDEVFPQAQLEQSK
370
1776.8
38-52

similar to F52H3.5

isoform 3 [Homo

sapiens]

CDC42-binding
gi|16357474|ref|NP_006026.2|
194314
IRPLNSEGTLNLLNCEPPR
371
2193.1
1503-1521

protein kinase beta

[Homo sapiens]

hect domain and
gi|7705931|ref|NP_057407.1|
116834
RTTEMMPVYLDLNK
372
1726.9
434-447

RLD S [Homo

sapiens]

hypothetical
gi|32526911|ref|NP_057697.2|
57535.6
QKCWQNGRVPK
373
1400.7
341-351

protein LOC51 313

isoform 2 [Homo

sapiens]

neuraminidase
gi|4557791|ref|NP_000425.1|
45449.3
DGVFCLLSDDHGASWR
374
1834.8
226-241

precursor [Homo

sapiens]

upstream binding
gi|7657671|ref|NP_055048.1|
89391.6
KPAQEGGK
375
814.44
394-401

transcription

factor, RNA

polymerase I

[Homo sapiens]

sarcolemma
gi|56550043|ref|NP_009090.2|
93187.1
EKGNNK
376
689.36
777-782

associated protein

[Homo sapiens]

immunoglobulin-
gi|28557777|ref|NP_787120.1|
57757
FGHPCSMLSSLGSEWER
377
2007.9
266-283

like domain

containing

receptor 1 [Homo

sapiens]

Janus kinase 3
gi|47157315|ref|NP_000206.2|
125083.7
EQGECLSLAVLDLARMAREQA
378
2644.3
158-180

[Homo sapiens]

QR

regulatory solute
gi|5730021|ref|NP_006502.1|
66771.3
DLGQGIQNSVTDRPETR
379
1885.9
523-539

carrier protein,

family 1, member

1 [Homo sapiens]

complement
gi|4502511|ref|NP_001728.1|
63156.8
AIEDYINEFSVR
380
1455.7
497-508

component 9

[Homo sapiens]

PREDICTED:
gi|113428147|ref|XP_038604.
202106.8
PTCELGDMSLLCVGVKK
381
1922.9
82-98

similar to Unc-13
10|

homolog A

(Muncl3-1) [Homo

sapiens]

apolipoprotein L1
gi|21735614|ref|NP_003652.21
43957.3
ILQADQEL
382
929.49
391-398

isoform a

precursor [Homo

sapiens]

complement
gi|4502503|ref|NP_000706.1|
67015
LSLEIEQLELQR
383
1470.8
574-585

component 4

binding protein,

alpha chain

precursor [Homo

sapiens]

cathelicidin
gi|39753970|ref|NP_004336.2|
19284.3
FALLGDFFR
384
1085.6
132-140

antimicrobial

peptide [Horno

sapiens]

ficolin 3 isoform 2
gi|27754778|ref|NP_775628.1|
31659.9
YGIDWASGR
385
1024.5
266-274

precursor [Homo

sapiens]

sterile alpha motif
gi|51339291|ref|NP_689916.2|
184522.8
SNFDETYIENWRNILK
386
2054.1
894-910

domain containing

9-like [Homo

sapiens]

OAF homolog
gi|30425438|ref|NP_848602.1|
30670.4
KPDGTLVSFTADFK
387
1525.8
63-76

[Homo sapiens]

gelsolin isoform b
gi|38044288|ref|NP_937895.1|
80622.8
EVQGFESATFLGYFK
388
1722.8
97-111

[Homo sapiens]

Example 3
Cancer-Related Peptides Are Not Necessarily Biomarkers

The above methods showed that a number of peptides previously known to be assosciated with colorectal cancer were not indicative of a disease state, and, thus, not useful as a biomarker. Examples include, alpha-1-antitrypsin precursor (alpha-1 protease inhibitor) (alpha-1-antiproteinase), follistatin-related protein 5 precursor (follistatin-like 5), sodium-D-glucose cotransporter (regulatory solute carrier protein, family 1, member 1), hypothetical protein DKFZp781M0386, alpha-1-acid glycoprotein 1 precursor (AGP 1) (Orosomucoid-1) (OMD 1), complement component C9 precursor that Contains complement component C9a and complement component C9b, hypothetical protein, immunoglobulin J chain, serum amyloid A-4 protein precursor (constitutively expressed serum amyloid A protein) (C-SAA), apolipoprotein A-II precursor (Apo-AII) (ApoA-II) that contains apolipoprotein A-II (1-76), IGKC protein, serum albumin precursor, complement factor B precursor (EC 3.4.21.47) (C3/C5 convertase) (properdin factor B) (glycine-rich beta glycoprotein) (GBG) (PBF2) that contains complement factor B Ba fragment and complement factor B Bb fragment, hemopexin precursor (Beta-1B-glycoprotein), intercellular adhesion molecule 5 precursor (ICAM-5) (telencephalin), receptor interacting protein kinase 5, isoform 2, Ig heavy chain V-III region TIL, probable ATP-dependent RNA helicase DDX43 (EC 3.6.1.-) (DEAD-box protein 43) (DEAD-box protein HAGE) (helical antigen), FLJ10748 protein, hypothetical protein DKFZp686J11235 (fragment), C219-reactive peptide (FLJ39207), Ig kappa chain V-II region RPMI 6410 precursor, Ig kappa chain V-I region AU, homeobox protein Hox-A4 (Hox-1D) (Hox-1.4), cullin-4B (CUL-4B), zinc finger protein ZFPM1 (zinc finger protein multitype 1) (friend of GATA protein 1) (friend of GATA-1) (FOG-1), two-pore calcium channel protein 2 (two pore segment channel 2), stonin-2 (stoned B), hypothetical protein FLJ45653, hypothetical protein DKFZp434A128, Ig kappa chain V-I region CAR, ras-related protein Rap-1A (GTP-binding protein smg-p21A) (ras-related protein Krev-1) (C21KG) (G-22K), hypothetical protein FLJ37300, hypothetical protein FLJ36006, mirror-image polydactyly gene 1 protein, gamma-tubulin complex component 3 (GCP-3) (spindle pole body protein Spc98 homolog) (hSpc98) (hGCP3) (h104p), HERV-W_—7q21.2 provirus ancestral Env polyprotein precursor (envelope polyprotein) (HERV-7q Envelope protein) (HERV-W envelope protein) (syncytin) (syncytin 1) (enverin) (Env-W) that contains surface protein (SU) and transmembrane protein (TM), low-density lipoprotein receptor-related protein 2 precursor (megalin) (glycoprotein 330) (gp330), 40S ribosomal protein S16, Nuclear pore complex protein Nup214 (nucleoporin Nup214) (214 kDa nucleoporin) (CAN protein), cadherin EGF LAG seven-pass G-type receptor 1 precursor (flamingo homolog 2) (hFmi2), and KIAA0425 protein (fragment).

BIOMARKERS FOR COLORECTAL CANCER

Information

Publication Number

Date Filed

Date Published

Inventors

Original Assignees

CPC

US Classifications

International Classifications

Abstract

Description

Claims

CROSS-REFERENCE TO RELATED APPLICATIONS

PCT Information

Provisional Applications (1)