Claims
- 1. Method for determining the presence of at least one target ligand, capable of competing with a ligand analogue conjugate for binding sites available on a ligand receptor, said ligand analogue conjugate comprising at least one ligand analogue coupled to a colloidal gold particle, in a fluid sample suspected of containing said target ligand comprising the steps of:
- a. contacting said fluid sample with said ligand analogue conjugate and said ligand receptor to form a homogeneous reaction mixture, the relative amounts of said ligand analogue conjugate and said ligand receptor being selected such that in the absence of said target ligand and subsequent to substantially equilibrium binding in said reaction mixture, substantially all of said ligand analogue conjugate is bound to said ligand receptor such that no unbound ligand analogue conjugate is detected as a result of the assay method;
- b. detecting unbound ligand analogue conjugates in said reaction mixture by visual means using at least one ligand receptor immobilized on a solid phase in at least one distinct locus;
- c. relating the detectable signal to the presence of said target ligand in said fluid sample.
- 2. Method for determining the presence of at least one target ligand, capable of competing with a ligand analogue conjugate for binding sites available on a ligand receptor, said ligand analogue conjugate comprising at least one ligand analogue coupled to a colloidal gold particle, in a fluid sample suspected of containing said target ligand, comprising the steps of:
- a. contacting said fluid sample with said ligand analogue conjugate and said ligand receptor to form a homogeneous reaction mixture, the relative amounts of said ligand analogue conjugate and said ligand receptor being selected such that in the absence of said target ligand and, subsequent to substantially equilibrium binding in said reaction mixture, substantially all of said ligand analogue conjugate is bound to said ligand receptor such that no unbound ligand analogue conjugate is detected as a result of the assay method;
- b. removing ligand receptor from said reaction mixture;
- c. detecting unbound ligand analogue conjugate in said reaction mixture by visual means using at least one ligand receptor immobilized on a solid phase in at least one distinct locus;
- d. relating the detectable signal to the presence of said target ligand in said fluid sample.
- 3. Method for determining the presence of at least one target ligand at or above at least one predetermined threshold ligand concentration, said target ligand capable of competing with a ligand analogue conjugate for binding sites available on a ligand receptor, said ligand analogue conjugate comprising at least one ligand analogue coupled to a colloidal gold particle, in a fluid sample suspected of containing said target ligand comprising the steps of:
- a. contacting said fluid sample with said ligand analogue conjugate and said ligand receptor to form a homogeneous reaction mixture;
- (i) the relative amounts of said ligand analogue conjugate and said ligand receptor being selected such that in the absence of said target ligand, and subsequent to substantially equilibrium binding in said reaction mixture, substantially all of said ligand analogue conjugate is bound to said ligand receptor such that no unbound ligand analogue conjugate is detected as a result of the assay method; and
- (ii) the relative and absolute amounts of ligand analogue conjugate and ligand receptor being selected such that at least one of said predetermined threshold ligand concentration is established below which no unbound ligand analogue conjugate is detected as a result of the assay method;
- b. detecting unbound ligand analogue conjugate in said reaction mixture by visual means using at least one ligand receptor immobilized on a solid phase in at least one distinct locus;
- c. relating the detectable signal to the amount of said target ligand at or above said predetermined threshold ligand concentration in said fluid sample.
- 4. Method for determining the presence of at least one target ligand at or above at least one predetermined threshold ligand concentration, said target ligand capable of competing with a ligand analogue conjugate for binding sites available on a ligand receptor, said ligand analogue conjugate comprising at least one ligand analogue coupled to a colloidal gold particle, in a fluid sample suspected of containing said target ligand, comprising the steps of:
- a. contacting said fluid sample with said ligand analogue conjugate and said ligand receptor to form a homogeneous reaction mixture:
- (i) the relative amounts of said ligand analogue conjugate and said ligand receptor being selected such that in the absence of said target ligand, and subsequent to substantially equilibrium binding in said reaction mixture, substantially all of said ligand analogue conjugate is bound to said ligand receptor such that no unbound ligand analogue conjugate is detected as a result of the assay method; and
- (ii) the relative and absolute amounts of ligand analogue conjugate an ligand receptor being selected such that at least one of said predetermined threshold ligand concentration is established below which no unbound ligand analogue conjugate is detected as a result of the assay method;
- b. removing ligand receptor from said reaction mixture;
- c. detecting unbound ligand analogue conjugate in said reaction mixture by visual means using at least one ligand receptor immobilized on a solid phase in at least one distinct locus;
- d. relating the detectable signal to the amount of said target ligand at or above said predetermined threshold ligand concentration in said fluid sample.
Parent Case Info
This application is a continuation of U.S. application Ser. No. 07/832,865, filed Feb. 6, 1992 (abandoned), and a continuation of U.S. application Ser. No. 07/463,150, filed Jan. 10, 1990 and issued as U.S. Pat. No. 5,089,391 on Feb. 18, 1992, which is a continuation in part of Ser. No. 07/295,568 filed Jan. 10, 1989 and issued as U.S. Pat. No. 5,028,535, which are hereby incorporated by reference and from which priority is claimed.
US Referenced Citations (27)
Foreign Referenced Citations (5)
Number |
Date |
Country |
0203238 |
Dec 1986 |
EPX |
0253464 |
Jan 1988 |
EPX |
0267006 |
May 1988 |
EPX |
2171999 |
Sep 1986 |
GBX |
8606170 |
Oct 1986 |
WOX |
Non-Patent Literature Citations (7)
Entry |
Lee, S. & & Liberti, P.A., Analytical Biochem., 166:41-48 (1987). |
Ekins, R.P., et al., "Theoretical Aspects of Saturation and Radio Immunoassay" in Radioiostopes in Mediciine: In Vitro Studies (Goswitz & Murphy eds.) U.S. Atomic Energy Comm'n, Oak Ridge, TN,9-100 (1968). |
Rodgers, R., et al., Clin. Chem. 24:95-100 (1976). |
Liu, D., et al., Clin. Toxicology, 25:527-538 (1987). |
Pluska, M.G., et al., BioTechniques, 4:272-283 (1986). |
Zettner, A., Clin. Chem. 19:699-705 (1973). |
Yalow, R.S. & Berson, S.A., "General Principles of Radio Immunoassay" in Radioisotopes in Medicine: In Vitro Studies (Goswitz & Murphy eds.), U.S. Atomic Energy Comm'n, Oak Ridge, TN 7-41 (1968). |
Related Publications (1)
|
Number |
Date |
Country |
|
463150 |
Jan 1990 |
|
Continuations (1)
|
Number |
Date |
Country |
Parent |
832865 |
Feb 1992 |
|
Continuation in Parts (1)
|
Number |
Date |
Country |
Parent |
295568 |
Jan 1989 |
|