Patent Application
20150197820
The Sequence Listing associated with this application is filed in electronic format via EFS-Web and hereby is incorporated by reference in its entirety into the specification. The name of the text file containing the Sequence Listing is 516154seqlist.txt. The size of the text file is 97.3 KB, and the text file was created on Mar. 30, 2015.
The present invention relates to novel marker sequences for inflammatory prostate diseases, prostate carcinoma, and the diagnostic use thereof together with a method for screening potential active substances for prostate diseases of this type by means of these marker sequences. Furthermore, the invention relates to a diagnostic device containing marker sequences of this type for inflammatory prostate diseases and prostate carcinoma, in particular a protein biochip and the use thereof.
Protein biochips are gaining increasing industrial importance in analysis and diagnosis as well as in pharmaceutical development. Protein biochips have become established as screening instruments.
The rapid and highly parallel detection of a multiplicity of specifically binding analysis molecules in a single experiment is rendered possible hereby. To produce protein biochips, it is necessary to have the required proteins available. For this purpose, in particular protein expression libraries have become established. The high throughput cloning of defined open reading frames is one possibility (Heyman, J. A., Cornthwaite, J., Foncerrada, L., Gilmore, J. R., Gontang, E., Hartman, K. J., Hernandez, C. L., Hood, R., Hull, H. M., Lee, W. Y., Marcil, R., Marsh, E. J., Mudd, K. M., Patino, M. J., Purcell, T. J., Rowland, J. J., Sindici, M. L. and Hoeffler, J. P., (1999) Genome-scale cloning and expression of individual open reading frames using topoisomerase I-mediated ligation. Genome Res, 9, 383-392; Kersten, B., Feilner, T., Kramer, A., Wehrmeyer, S., Possling, A., Witt, I., Zanor, M. I., Stracke, R., Lueking, A., Kreutzberger, J., Lehrach, H. and Cahill, D. J. (2003) Generation of Arabidopsis protein chip for antibody and serum screening. Plant Molecular Biology, 52, 999-1010; Reboul, J., Reboul, J., Vaglio, P., Rual, J. F., Lamesch, P., Martinez, M., Armstrong, C M., Li, S., Jacotot, L., Bertin, N., Janky, R., Moore, T., Hudson, J. R., Jr., Hartley, J. L., Brasch, M. A., Vandenhaute, J., Boulton, S., Endress, G. A., Jenna, S., Chevet, E., Papasotiropoulos, V., Tolias, P. P., Ptacek, J., Snyder, M., Huang, R., Chance, M. R., Lee, H., Doucette-Stamm, L., Hill, D. E. and Vidal, M. (2003) C. elegans ORFeome Version 1.1: experimental verification of the genome annotation and resource for proteome-scale protein expression. Nat Genet, 34, 35-41.; Walhout, A. J., Temple, G. F., Brasch, M. A., Hartley, J. L., Lorson, M. A., van den Heuvel, S. and Vidal, M. (2000) GATEWAY recombinational cloning: application to the cloning of large numbers of open reading frames or ORFeomes. Methods Enzymol, 328, 575-592). However, an approach of this type is strongly connected to the progress of the genome sequencing projects and the annotation of these gene sequences. Furthermore, the determination of the expressed sequence can be ambiguous due to differential splicing processes. This problem may be circumvented by the application of cDNA expression libraries (Büssow, K., Cahill, D., Nietfeld, W., Bancroft, D., Scherzinger, E., Lehrach, H. and Walter, G. (1998) A method for global protein expression and antibody screening on high-density filters of an arrayed cDNA library. Nucleic Acids Research, 26, 5007-5008; Büssow, K., Nordhoff, E., Lübbert, C, Lehrach, H. and Walter, G. (2000) A human cDNA library for high-throughput protein expression screening. Genomics, 65, 1-8; Holz, C, Lueking, A., Bovekamp, L., Gutjahr, C, Bolotina, N., Lehrach, H. and Cahill, D. J. (2001) A human cDNA expression library in yeast enriched for open reading frames. Genome Res, 11, 1730-1735; Lueking, A., Holz, C, Gotthold, C, Lehrach, H. and Cahill, D. (2000) A system for dual protein expression in Pichia pastoris and Escherichia coli, Protein Expr. Purif., 20, 372-378). The cDNA of a particular tissue is hereby cloned into a bacterial or an eukaryotic expression vector, such as, e.g., yeast. The vectors used for the expression are generally characterized in that they carry inducible promoters that may be used to control the time of protein expression. Furthermore, expression vectors have sequences for so-called affinity epitopes or affinity proteins, which on the one hand permit the specific detection of the recombinant fusion proteins by means of an antibody directed against the affinity epitope, and on the other hand the specific purification via affinity chromatography (IMAC) is rendered possible.
For example, the gene products of a cDNA expression library from human fetal brain tissue in the bacterial expression system Escherichia coli were arranged in high-density format on a membrane and could be successfully screened with different antibodies. It was possible to show that the proportion of full-length proteins is at least 66%. Additionally, the recombinant proteins from the library could be expressed and purified in a high-throughput manner (Braun P., Hu, Y., Shen, B., Halleck, A., Koundinya, M., Harlow, E. and LaBaer, J. (2002) Proteome-scale purification of human proteins from bacteria. Proc Natl Acad Sci U S A, 99, 2654-2659; Büssow (2000) supra; Lueking, A., Horn, M., Eickhoff, H., Büssow, K., Lehrach, H. and Walter, G. (1999) Protein microarrays for gene expression and antibody screening. Analytical Biochemistry, 270, 103-111). Protein biochips of this type based on cDNA expression libraries are in particular the subject matter of WO 99/57311 and WO 99/57312.
Furthermore, in addition to antigen-presenting protein biochips, antibody-presenting arrangements are likewise described (Lal et al (2002) Antibody arrays: An embryonic but rapidly growing technology, DDT, 7, 143-149; Kusnezow et al. (2003), Antibody microarrays: An evaluation of production parameters, Proteomics, 3, 254-264).
However, there is a great need to provide indication-specific diagnostic devices, such as a protein biochip.
The laboratory parameters include acid phosphatase (AP) and prostate-specific antigen (PSA) for diagnosing prostate carcinoma. Above all, PSA currently has a high importance in diagnostics. It is specific for the prostate, but not for a tumor disease, but rather can also be elevated in the event of inflammation, benign prostate hyperplasia, urine retention, or without an obvious reason. A value over 4 ng/mL already requires clarification.
The object of the present invention is therefore to provide improved marker sequences and the diagnostic use thereof for the treatment of inflammatory prostate diseases up to prostate carcinoma.
The provision of specific marker sequences permits a reliable diagnosis and stratification of patients with inflammatory prostate diseases up to prostate carcinoma, in particular by means of a protein biochip.
The invention therefore relates to the use of marker sequences for the diagnosis of inflammatory prostate diseases up to prostate carcinoma, wherein at least one marker sequence of a cDNA selected from the group SEQ ID NOS: 1-174 or respectively a protein coding therefor or respectively a partial sequence or fragment thereof (hereinafter: marker sequences according to the invention) is determined on or from a patient to be examined.
It was possible to identify the marker sequences according to the invention by means of differential screening of samples from healthy test subjects with patient samples with inflammatory prostate diseases, prostate carcinoma.
For the first time, these marker sequences according to the invention could be identified by means of protein biochips (see examples) hereby.
The term “inflammatory prostate diseases up to prostate carcinoma” comprises a group of diseases from prostatitis up to the chronic forms of all prostate inflammations and the establishment thereof as prostate cancer or prostate carcinoma (definition, e.g., according to Pschyrembel, de Gruyter, 261st edition (2007), Berlin).
In a further embodiment at least 2 to 5 or 10, preferably 30 to 50 marker sequences, or 50 to 100 or more marker sequences are determined on or from a patient to be examined.
In a further embodiment of the invention, the marker sequences according to the invention can likewise be combined, supplemented, fused, or expanded likewise with known biomarkers for this indication.
In a preferred embodiment, the determination of the marker sequences is carried out outside the human body and the determination is carried out in an ex vivo/in vitro diagnosis.
In a further embodiment of the invention, the invention relates to the use of marker sequences as diagnostic agents, wherein at least one marker sequence of a cDNA is selected from the group SEQ ID NOS: 1-174 or respectively a protein coding therefor or respectively a partial sequence or fragment thereof.
Furthermore, the invention relates to a method for the diagnosis of inflammatory prostate diseases up to prostate carcinoma, wherein a.) at least one marker sequence of a cDNA selected from the group SEQ ID NOS: 1-174 or respectively a protein coding therefor or respectively a partial sequence or fragment thereof is applied to a solid support and b.) is brought into contact with body fluid or tissue extract of a patient and c.) the detection of an interaction of the body fluid or tissue extract with the marker sequences from a.) is carried out.
The invention therefore likewise relates to diagnostic agents for the diagnosis of inflammatory prostate diseases up to prostate carcinoma respectively selected from the group SEQ ID NOS: 1-174 or respectively a protein coding therefor or respectively a partial sequence or fragment thereof.
In a particularly preferred embodiment, the marker sequences SEQ ID NOS: 136, 40, 127, 83, 16, 82, 88, 152, 130, 138, 2, 12, 113, 20, 173, 33, 172, 52, 43, 91, 1, 32, 86, 27, 105 are preferred in this order.
The detection of an interaction of this type can be carried out, for example, by a probe, in particular by an antibody.
The invention therefore likewise relates to the object of providing a diagnostic device or an assay, in particular a protein biochip, which permits a diagnosis or examination for inflammatory prostate diseases up to prostate carcinoma.
Furthermore, the invention relates to a method for the stratification, in particular risk stratification and/or therapy control of a patient with inflammatory prostate diseases up to prostate carcinoma, wherein at least one marker sequence of a cDNA selected from the group SEQ ID NOS: 1-174 or respectively a protein coding therefor is determined on a patient to be examined.
Furthermore, the stratification of the patients with inflammatory prostate diseases up to prostate carcinoma in new or established subgroups of inflammatory prostate diseases up to prostate carcinoma is also covered, as well as the expedient selection of patient groups for the clinical development of novel therapeutic agents. The term therapy control likewise covers the allocation of patients to responders and non-responders regarding a therapy or the therapy course thereof.
“Diagnosis” for the purposes of this invention means the positive determination of inflammatory prostate diseases up to prostate carcinoma by means of the marker sequences according to the invention as well as the assignment of the patients to inflammatory prostate diseases up to prostate carcinoma. The term diagnosis covers medical diagnostics and examinations in this regard, in particular in-vitro diagnostics and laboratory diagnostics, likewise proteomics and nucleic acid blotting. Further tests can be necessary to be sure and to exclude other diseases. The term diagnosis therefore likewise covers the differential diagnosis of inflammatory prostate diseases, prostate carcinoma by means of the marker sequences according to the invention and the prognosis of inflammatory prostate diseases and prostate carcinoma.
“Stratification or therapy control” for the purposes of this invention means that the method according to the invention renders possible decisions for the treatment and therapy of the patient, whether it is the hospitalization of the patient, the use, effect and/or dosage of one or more drugs, a therapeutic measure or the monitoring of a course of the disease and the course of therapy or etiology or classification of a disease, e.g., into a new or existing subtype or the differentiation of diseases and the patients thereof.
In a further embodiment of the invention, the term “stratification” covers in particular the risk stratification with the prognosis of an outcome of a negative health event.
Within the scope of this invention, “patient” means any test subject—human or mammal—with the proviso that the test subject is tested for inflammatory prostate diseases up to prostate carcinoma.
The term “marker sequences” for the purposes of this invention means that the cDNA or the polypeptide or protein that can be respectively obtained therefrom are significant for inflammatory prostate diseases, prostate carcinoma. For example, the cDNA or the polypeptide or protein that can be respectively obtained therefrom can exhibit an interaction with substances from the body fluid or tissue extract of a patient with inflammatory prostate diseases, prostate carcinoma (e.g., antigen (epitope)/antibody (paratope) interaction). For the purposes of the invention “wherein at least one marker sequence of a cDNA selected from the group SEQ ID NOS: 1-174 or respectively a protein coding therefor or respectively a partial sequence or fragment thereof is determined on a patient to be examined” means that an interaction between the body fluid or tissue extract of a patient and the marker sequences according to the invention is detected. An interaction of this type is, e.g., a bond, in particular a binding substance on at least one marker sequence according to the invention or in the case of a cDNA the hybridization with a suitable substance under selected conditions, in particular stringent conditions (e.g., such as usually defined in J. Sambrook, E. F. Fritsch, T. Maniatis (1989), Molecular cloning: A laboratory manual, 2nd Edition, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, USA or Ausubel, “Current Protocols in Molecular Biology,” Green Publishing Associates and Wiley Interscience, N.Y. (1989)). One example of stringent hybridization conditions is: hybridization in 4×SSC at 65° C. (alternatively in 50% formamide and 4×SSC at 42° C.), followed by several washing steps in 0.1×SSC at 65° C. for a total of approximately one hour. An example of less stringent hybridization conditions is hybridization in 4×SSC at 37° C., followed by several washing steps in 1×SSC at room temperature.
According to the invention, substances of this type are constituents of a body fluid, in particular blood, whole blood, blood plasma, blood serum, patient serum, urine, cerebrospinal fluid, synovial fluid, or of a tissue extract of the patient.
In a further embodiment of the invention, however, the marker sequences according to the invention can be present in a significantly higher or lower expression rate or concentration that indicates inflammatory prostate diseases, prostate carcinoma. The relative sick/healthy expression rates of the marker sequences for inflammatory prostate diseases, prostate carcinoma according to the invention are hereby determined by means of proteomics or nucleic acid blotting.
In a further embodiment of the invention, the marker sequences have a recognition signal that is addressed to the substance to be bound (e.g., antibody, nucleic acid). It is preferred according to the invention that for a protein the recognition signal is an epitope and/or a paratope and/or a hapten and for a cDNA is a hybridization or binding region.
The marker sequences according to the invention are the subject matter of Table A and can be clearly identified by the respectively cited database entry (also by means of the Internet: ncbi.nlm.nih.gov) (see in Table A: accession no. there), see also the associated sequence protocol.
The invention therefore also relates to the full-length sequences of the markers according to the invention, as defined in Table 1 via the known database entry according to Table A.
Therefore, the invention also comprises analogous embodiments of the sequences of the known database entries listed in Table A to the marker sequences SEQ ID NOS: 1-174, such as, e.g., described in the claims, since the SEQ ID NOS: 1-174 according to the invention in turn represent partial sequences, at least with high homology. The specific marker sequences SEQ ID NOS: 1-174 are preferred according to the invention, however.
Furthermore, the following sequences of the known database entries listed in Table A are preferred: gi:61102726, gi:23308566, gi:56117827 (SEQ ID NO: 175), gi:40789263, gi:13375724, gi:40354199, gi:41393564, gi:7705400, gi:57617038, gi:63082031, gi:113413768, gi:12751496, gi:50878292, gi:14251213, gi:94536841, gi:22035555, gi:90903237, gi:30089990, gi:24308256, gi:44917603, gi:13402448, gi:22027484, gi:41352062, gi:19743568, and gi:47717133.
According to the invention, the marker sequences also cover those modifications of the cDNA sequence and the corresponding amino acid sequence as chemical modification, such as citrullination, acetylation, phosphorylation, glycosylation or poly(A) strand and other modifications known to one skilled in the art.
In a further embodiment of the invention, partial sequences or fragments of the marker sequences according to the invention are likewise comprised. In particular those partial sequences that have an identity of 95%, 90%, in particular 80% or 70% with the marker sequences according to the invention.
Partial sequences are also sequences of the type which have 50 to 100 nucleotides, 70-120 nucleotides of a sequence of the SEQ ID NOS: 1-174, or peptides obtainable therefrom.
In a further embodiment, the respective marker sequence can be represented in different quantities in one more regions on a solid support. This permits a variation of the sensitivity. The regions can have respectively a totality of marker sequences, i.e., a sufficient number of different marker sequences, in particular 2 to 5 or 10 or more and optionally more nucleic acids and/or proteins, in particular biomarkers. However, at least 96 to 25,000 (numerical) or more from different or identical marker sequences and further nucleic acids and/or proteins, in particular biomarkers are preferred. Furthermore preferred are more than 2,500, in particular preferred 10,000 or more different or identical marker sequences and optionally further nucleic acids and/or proteins, in particular biomarkers.
Another object of the invention relates to an arrangement of marker sequences containing at least one marker sequence of a cDNA selected from the group SEQ ID NOS: 1-174 or respectively a protein coding therefor. Preferably, the arrangement contains at least 2 to 5 or 10, preferably 30 to 50 marker sequences, or 50 to 100 or more marker sequences.
Within the scope of this invention, “arrangement” is synonymous with “array,” and if this “array” is used to identify substances on marker sequences, this is to be understood to be an “assay” or diagnostic device. In a preferred embodiment, the arrangement is designed such that the marker sequences represented on the arrangement are present in the of a grid on a solid support. Furthermore, those arrangements are preferred that permit a high-density arrangement of protein binders and the marker sequences are spotted. Such high-density spotted arrangements are disclosed, for example, in WO 99/57311 and WO 99/57312 and can be used advantageously in a robot-supported automated high-throughput method.
Within the scope of this invention, however, the term “assay” or diagnostic device likewise comprises those embodiments of a device, such as ELISA, bead-based assay, line assay, Western Blot, immunochromatographic methods (e.g., so-called lateral flow immunoassays, or similar immunological single or multiplex detection measures. A protein biochip in terms of this invention is the systematic arrangement of proteins on a solid support.
The marker sequences of the arrangement are fixed on a solid support, but preferably spotted or immobilized even printed on, i.e. applied in a reproducible manner. One or more marker sequences can be present multiple times in the totality of all marker sequences and present in different quantities based on one spot. Furthermore, the marker sequences can be standardized on the solid support (i.e., by means of serial dilution series of, e.g., human globulins as internal calibrators for data normalization and quantitative evaluation).
The invention therefore relates to an assay or a protein biochip comprising an arrangement containing marker sequences according to the invention.
In a further embodiment, the marker sequences are present as clones. Clones of this type can be obtained, for example, by means of a cDNA expression library according to the invention (Büssow et al. 1998 (supra)). In a preferred embodiment, such expression libraries containing clones are obtained using expression vectors from a cDNA expression library comprising the cDNA marker sequences. These expression vectors preferably contain inducible promoters. The induction of the expression can be carried out, e.g., by means of an inductor, such as IPTG. Suitable expression vectors are described in Terpe et al. (Terpe T Appl Microbiol Biotechnol. 2003 January; 60(5): 523-33).
One skilled in the art is familiar with expression libraries, they can be produced according to standard works, such as Sambrook et al, “Molecular Cloning, A laboratory handbook, 2nd edition (1989), CSH press, Cold Spring Harbor, N.Y. Expression libraries are also preferred which are tissue-specific (e.g., human tissue, in particular human organs). Furthermore included according to the invention are expression libraries that can be obtained by exon-trapping. A synonym for expression library is expression bank. Also preferred are protein biochips or corresponding expression libraries that do not exhibit any redundancy (so-called: UNICLONE® library) and that may be produced, for example, according to the teachings of WO 99/57311 and WO 99/57312. These preferred UNICLONE libraries have a high portion of non-defective fully expressed proteins of a cDNA expression library.
Within the context of this invention, the clones can also be, but not limited to, transformed bacteria, recombinant phages, or transformed cells from mammals, insects, fungi, yeasts, or plants.
The clones are fixed, spotted, or immobilized on a solid support.
The invention therefore relates to an arrangement wherein the marker sequences are present as clones.
Additionally, the marker sequences can be present in the respective form of a fusion protein, which contains, for example, at least one affinity epitope or tag. The tag may be one such as contains c-myc, his tag, arg tag, FLAG, alkaline phosphatase, VS tag, T7 tag or strep tag, HAT tag, NusA, S tag, SBP tag, thioredoxin, DsbA, a fusion protein, preferably a cellulose-binding domain, green fluorescent protein, maltose-binding protein, calmodulin-binding protein, glutathione S-transferase, or lacZ.
In all of the embodiments, the term “solid support” covers embodiments such as a filter, a membrane, a magnetic or fluorophore-labeled bead, a silica wafer, glass, metal, ceramics, plastics, a chip, a target for mass spectrometry, or a matrix. However, a filter is preferred according to the invention.
As a filter, furthermore PVDF, nitrocellulose, or nylon is preferred (e.g., Immobilon P Millipore, Protran Whatman, Hybond N+ Amersham).
In another preferred embodiment of the arrangement according to the invention, the arrangement corresponds to a grid with the dimensions of a microtiter plate (8-12 wells strips, 96 wells, 384 wells, or more), a silica wafer, a chip, a target for mass spectrometry, or a matrix.
In a further embodiment, the invention relates to an assay or a protein biochip for identifying and characterizing a substance for inflammatory prostate diseases, prostate carcinoma, characterized in that an arrangement or assay according to the invention is a.) brought into contact with at least one substance to be tested and b.) a binding success is detected.
Furthermore, the invention relates to a method for identifying and characterizing a substance for inflammatory prostate diseases, prostate carcinoma, characterized in that an arrangement or assay according to the invention is a.) brought into contact with at least one substance to be tested and b.) a binding success is detected.
The substance to be tested can be any native or non-native biomolecule, a synthetic chemical molecule, a mixture, or a substance library.
After the substance to be tested contacts a marker sequence, the binding success is evaluated, which, for example, is carried out using commercially available image analyzing software (GenePix Pro (Axon Laboratories), Aida (Ray test), ScanArray (Packard Bioscience)).
The visualization of protein-protein interactions according to the invention (e.g., protein on marker sequence, as antigen/antibody) or corresponding “means for detecting the binding success” can be performed, for example, using fluorescence labeling, biotinylation, radioisotope labeling, or colloid gold or latex particle labeling in the usual way. A detection of bound antibodies is carried out with the aid of secondary antibodies, which are labeled with commercially available reporter molecules (e.g., Cy, Alexa, Dyomics, FITC, or similar fluorescent dyes, colloidal gold or latex particles), or with reporter enzymes, such as alkaline phosphatase, horseradish peroxidase, etc., and the corresponding colorimetric, fluorescent, or chemiluminescent substrates. Readout is conducted, e.g., using a microarray laser scanner, a CCD camera, or visually.
In a further embodiment, the invention relates to a drug/active substance or prodrug developed for inflammatory prostate diseases, prostate carcinoma and obtainable through the use of the assay or protein biochip according to the invention.
The invention therefore likewise relates to the use of an arrangement according to the invention or an assay for screening active substances for inflammatory prostate diseases, prostate carcinoma.
In a further embodiment, the invention therefore likewise relates to a target for the treatment and therapy of inflammatory prostate diseases, prostate carcinoma respectively selected from the group SEQ ID NOS: 1-174 or a protein respectively coding therefor.
In a further embodiment, the invention likewise relates to the use of the marker sequences according to the invention, preferably in the form of an arrangement, as an affinity material for carrying out an apheresis or in the broadest sense a blood lavage, wherein substances from body fluids of a patient with inflammatory prostate diseases, prostate carcinoma, such as blood or plasma, bind to the marker sequences according to the invention and consequently can be selectively withdrawn from the body fluid.
Ten or more patient samples were individually screened against a cDNA expression library. The expression clones specific to inflammatory prostate diseases, prostate carcinoma were determined through a comparison with ten or more healthy samples. The identity of the marker sequences was determined by DNA sequencing.
In the scope of the biomarker identification, various bioinformatic analyses are performed. For each serum, reactivities against approximately 2000 different antigens are measured by means of microarray. These data are used for a ranking of the spotted antigens with respect to their differentiation capability between healthy and diseased sera. This analysis is performed by means of the non-parameterized Mann-Whitney test on normalized intensity data. An internal standard which is also spotted on each chip is used for the normalization. Since a p value is calculated for each antigen, methods are used for correction of the multiple test. As a very conservative approach, a Bonferroni direction is performed and the less restrictive false discovery rate (FDR) according to Benjamini & Hochberg is additionally calculated. Furthermore, the data are used for classification of the sera. Different multivariate methods are used hereby. These are methods from statistical learning methods such as support vector machines (SVM), neural networks, or classification trees, as well as a threshold value method, which is capable of both classification and also visual representation of the data.
To avoid overfitting, a 10-fold cross-validation of the data is performed.
Homo sapiens chromosome 8 open reading frame 33
Homo sapiens solute carrier family 27 (fatty acid
Homo sapiens ARP1 actin-related protein 1 homolog A,
Homo sapiens family with sequence similarity 77,
Homo sapiens chromosome 14 open reading frame 138
Homo sapiens cytochrome P450, family 27, subfamily A,
Homo sapiens spinster (SPIN1), mRNA
Homo sapiens heterogeneous nuclear ribonucleoprotein
Homo sapiens DEAD (Asp-Glu-Ala-Asp) box
Homo sapiens ribosomal protein S25 (RPS25), mRNA
Homo sapiens chromosome 20 genomic contig,
Homo sapiens ribosomal protein L 18a (RPL 18A),
Homo sapiens ribosomal protein, large, P1 (RPLP1),
Homo sapiens FK506 binding protein 1 A, 12kDa
Homo sapiens ribosomal protein L7a (RPL7A), mRNA
Homo sapiens TRAF family member-associated NFKB
Homo sapiens guanine nucleotide binding protein (G
Homo sapiens SWI/SNF related, matrix associated,
Homo sapiens homer homolog 3 (Drosophila)
Homo sapiens solute carrier family 25 (mitochondrial
Homo sapiens RAS, dexamethasone-induced 1
Homo sapiens BRF1 homolog, subunit of RNA
Homo sapiens LIM domain containing 2 (LIMD2), mRNA
Homo sapiens glutathione transferase zeta 1
Homo sapiens opioid receptor, sigma 1 (OPRS1),
Homo sapiens phosphatidylinositol glycan anchor
Homo sapiens RNA binding motif protein 10 (RBM1 0),
Homo sapiens carbohydrate (N-acetylglucosamine 6-0)
Homo sapiens asparaQinase like 1 (ASRGL 1), mRNA
Homo sapiens DnaJ (Hsp40) homolog, subfamily B,
Homo sapiens formin binding protein 4 (FNBP4), mRNA
Homo sapiens KIAA1576 protein (KIAA1576), mRNA
Homo sapiens Ras association (RaIGDS/AF-6) domain
Homo sapiens Kv channel interacting protein 1
Homo sapiens isocitrate dehydrogenase 2 (NADP+),
Homo sapiens serologically defined colon cancer
Homo sapiens CCAAT/enhancer binding protein
Homo sapiens chromosome 10 genomic contig,
Homo sapiens adducin 1 (alpha) (ADD1), transcript
Homo sapiens adducin 1 (alpha) (ADD1), transcript
Homo sapiens acid phosphatase 1, soluble (ACP1),
Homo sapiens histidyl-tRNA synthetase 2 (HARS2),
Homo sapiens axin 1 (AXIN1), transcript variant 1,
Homo sapiens sterile alpha motif domain containing 14
Homo sapiens chromosome 10 open reading frame 13
Homo sapiens splicing factor, arginine/serine-rich 1
Homo sapiens WD repeat domain 54 (WDR54), mRNA
Homo sapiens hypothetical protein DKFZp434G156
Homo sapiens protein kinase, interferon-inducible
Homo sapiens serpin peptidase inhibitor, Glade H (heat
Homo sapiens erythrocyte membrane protein band 4.1-
Homo sapiens splicing factor 4 (SF4), mRNA
Homo sapiens activating transcription factor 4 (tax-
Homo sapiens protein disulfide isomerase family A,
Homo sapiens phospholipase C, gamma 1 (PLCG1),
Homo sapiens RAS-like, family 11, member B (RASL11
Homo sapiens dehydrogenase/reductase (SDR family)
Homo sapiens family with sequence similarity 100,
Homo sapiens phosphatidylinositol 3,4,5-trisphosphate-
Homo sapiens chromosome 10 genomic contig,
Homo sapiens chromosome 10 genomic contig,
Homo sapiens chromatin modifying protein 2A
Homo sapiens alpha-2-glycoprotein 1, zinc (AZGP1),
Homo sapiens NAOH dehydrogenase (ubiquinone) Fe-S
Homo sapiens amyloid beta (A4) precursor protein-
Homo sapiens splNryanodine receptor domain and
Homo sapiens protein (peptidylprolyl cis/trans
Homo sapiens AOP-ribosylation factor-like 8A (ARL8A),
Homo sapiens fibroblast growth factor (acidic)
Homo sapiens small glutamine-rich tetratricopeptide
Homo sapiens TPX2, microtubule-associated, homolog
Homo sapiens hypothetical protein MGC11257
Homo sapiens ubiquitin specific peptidase 4 (proto-
Homo sapiens p300/CBP-associated factor (PCAF),
Homo sapiens phosphofructokinase, platelet (PFKP),
Homo sapiens vacuolar protein sorting 35 (yeast)
Homo sapiens inositol 1 ,3,4-triphosphate 5/6 kinase
Homo sapiens OEAH (Asp-Glu-Ala-His) box polypeptide
Homo sapiens NGFI-A binding protein 2 (EGR1 binding
Homo sapiens SLiT-ROBO Rho GTPase activating
Homo sapiens insulin-like growth factor binding protein
Homo sapiens Sjogren's syndrome nuclear autoantigen
Homo sapiens small nuclear ribonucleoprotein
Homo sapiens triple functional domain (PTPRF
Homo sapiens methylmalonyl Coenzyme A mutase
Homo sapiens Norrie disease (pseudoglioma) (NOP),
Homo sapiens TBC1 domain family, member 9B (with
Homo sapiens coiled-coil-helix-coiled-coil-helix domain
Homo sapiens chromosome 11 open reading frame 31
Homo sapiens aconitase 2, mitochondrial (AC02),
Homo sapiens protein kinase, AMP-activated, gamma 1
Homo sapiens protein kinase N1 (PKN1), transcript
Homo sapiens acetylserotonin O-methyltransferase-like
Homo sapiens CDC-like kinase 2 (CLK2), transcript
Homo sapiens RNA binding motif protein 15 (RBM15),
Homo sapiens golgi associated, gamma adaptin ear
Homo sapiens hypothetical protein FLJ1 0154 (FLJ1
Homo sapiens ELAV (embryonic lethal, abnormal vision,
Homo sapiens chromosome 14 open reading frame 131
Homo sapiens RNA binding motif protein 5 (RBM5),
Homo sapiens complement component 4B (Childo blood
Homo sapiens tripartite motif-containing 45 (TRIM45),
Homo sapiens chromosome 5 genomic contig,
Homo sapiens chromosome 8 genomic contig,
Homo sapiens chromosome 8 genomic contig,
Homo sapiens chromosome 16 genomic contig,
Homo sapiens chromosome 16 genomic contig,
Homo sapiens chromosome 17 genomic contig,
Homo sapiens chromosome 21 genomic contig,
Homo sapiens pim-3 oncogene (PIM3), mRNA
Homo sapiens melanoma antigen family D, 1
Homo sapiens RAB11 family interacting protein 4 (class
Homo sapiens valyl-tRNA synthetase like (VARSL),
Homo sapiens ubiquitin associated domain containing 1
Homo sapiens transcription elongation factor A (SII)-like
Homo sapiens speckle-type POZ protein (SPOP),
Homo sapiens speckle-type POZ protein (SPOP),
Homo sapiens adenomatosis polyposis coli 2 (APC2),
Homo sapiens tubulin tyrosine ligase-like family,
Homo sapiens AFG3 ATPase family gene 3-like 2
Homo sapiens zyxin (ZYX), transcript variant 2, mRNA
Homo sapiens spectrin, beta, non-erythrocytic 2
Homo sapiens ring finger protein 113A (RNF113A),
Homo sapiens 3-phosphoinositide dependent protein
Homo sapiens La ribonucleoprotein domain family,
Homo sapiens histone deacetylase 5 (HOAC5),
Homo sapiens p53-associated parkin-like cytoplasmic
Homo sapiens IQ motif and WO repeats 1 (IQW01),
Homo sapiens chromosome 1 open reading frame 131
Homo sapiens ATPase, Class II, type 9A (ATP9A),
Homo sapiens lipopolysaccharide-induced TNF factor
Homo sapiens membrane associated guanylate kinase,
Homo sapiens zinc finger protein 12 (ZNF12), mRNA
Homo sapiens AOP-ribosylation factor 1 (ARF1),
Homo sapiens family with sequence similarity 50,
Homo sapiens ribosomal protein S3A (RPS3A), mRNA
Homo sapiens asparaginyl-tRNA synthetase (NARS),
Homo sapiens polyglutamine binding protein 1
Homo sapiens upstream binding transcription factor,
Homo sapiens valosin-containing protein (VCP), mRNA
Homo sapiens HOCMA18P protein (HOCMA18P),
Homo sapiens chromosome 9 open reading frame 78
Homo sapiens endoplasmic reticulum protein 29
Homo sapiens ubiquitin-binding protein homolog
Homo sapiens heme oxygenase (decycling) 2 (HMOX2),
Homo sapiens kinesin family member 21 B (KIF21 B),
Homo sapiens CaM kinase-like vesicle-associated
Homo sapiens microtubule-associated protein 2
Homo sapiens chromosome 1 genomic contig,
Homo sapiens chromosome 1 genomic contig,
Homo sapiens chromosome 2 genomic contig, alternate
Homo sapiens chromosome 6 genomic contig, alternate
Homo sapiens chromosome 6 genomic contig, alternate
Homo sapiens chromosome 8 genomic contig, alternate
Homo sapiens chromosome 14 genomic contig,
Homo sapiens chromosome 19 genomic contig,
Homo sapiens chromosome X genomic contig,
Homo sapiens chromosome X genomic contig,
Homo sapiens PRP38 pre-mRNA processing factor 38
Homo sapiens protein tyrosine phosphatase, non-
Homo sapiens glutathione peroxidase 4 (phospholipid
Homo sapiens ribose 5-phosphate isomerase A (ribose
Homo sapiens zuotin related factor 1 (ZRF1), mRNA
SEQUENCE OF Homo sapiens speckle-type POZ protein (SPOP), transcript variant 3, mRNA from Genbank Accession No. gi:56117827 (SEQ ID NO: 175):
| Number | Date | Country | Kind |
|---|---|---|---|
| 102008031699.7 | Jul 2008 | DE | national |
This application is a continuation application of U.S. patent application Ser. No. 13/860,345, filed Apr. 10, 2013, which is incorporated by reference herein in its entirety, which was a continuation application of U.S. patent application Ser. No. 13/002,098, filed Mar. 7, 2011, which is a national stage application (under 35 U.S.C. 371) of PCT/EP2009/058534 filed Jul. 6, 2009, which claims benefit of German Application No. 102008031699.7 filed Jul. 4, 2008. The entire contents of each of these applications are hereby incorporated by reference herein in their entireties.
| Number | Date | Country | |
|---|---|---|---|
| Parent | 13860345 | Apr 2013 | US |
| Child | 14672334 | US | |
| Parent | 13002098 | Mar 2011 | US |
| Child | 13860345 | US |
