Claims for Patent: 4,935,339

Summary for Patent: 4,935,339

Title:	Delayed solid phase immunologic assay
Abstract:	A method for the determination of a substance present in a sample, which comprises: (a) contacting a sample containing a substance with: (i) a first immunological binding partner to the substance, wherein the first immunological binding partner is a monoclonal antibody bound to biotin or a biotin-binding protein, (ii) a second immunological binding partner to the substance, wherein the second immunological binding partner is a detectably labeled region specific polyclonal antibody, and (iii) a biotin binding protein or biotin bound to a carrier; (b) incubating the components of step (a) for a period of time and under conditions sufficient to form an immune complex between the substance, the first immunological binding partner, the second immunological binding partner, and the carrier; (c) separating the carrier from the sample; and (d) determining the detectably labeled second immunological binding partner in either the sample or the carrier; wherein the reaction between the first immunological binding partner and the carrier occurs after, or substantially contemporaneously with, formation of an immune complex between the first immunological binding partner and the substance.
Inventor(s):	Zahradnik; Richard (San Juan Capistrano, CA)
Assignee:	Nichols Institute Diagnostics (San Juan Capistrano, CA)
Application Number:	06/731,470
Patent Claims:	1. A method of preparing a region specific polyclonal antibody to a substance capable of being bound by antibody and having at least two different epitopes, comprising the steps of: (a) binding said substance to a carrier via one or more monoclonal antibodies specific for an epitope on said substance, said monoclonal antibody and said substance being bound to said carrier under conditions which prevent their elution in step (d); (b) binding a polyclonal antibody specific for said substance to said carrier-bound substance; (c) washing said bound polyclonal antibody; (d) eluting said bound polyclonal antibody from said substance under conditions such that said monoclonal antibody and said substance remain bound to said carrier; and (e) recovering said region specific polyclonal antibody. 2. A method for the determination of a substance present in a sample, which comprises: (a) contacting said sample containing a substance having binding sites thereon capable of binding to at least two different immunological binding partners therefor with: (i) a first immunological binding partner to said substance, said partner being bound to biotin or a biotin binding protein; (ii) a detactably labeled, second immunological binding partner to said substance; and (iii) a carrier bound to a biotin binding protein or to biotin; wherein said first immunological binding partner is a monoclonal antibody and said immunological binding partner is a region specific polyclonal antibody prepared by the method of claim 1; (b) incubating the components of step (a) for a period of time and under conditions sufficient to form an immune complex between said substance, said first immunological binding partner, said second immunological binding partner, and said carrier; (c) separating the immune complex-carrier from the sample; and (d) determining the presence of such substance by determining the detactably labeled immunological binding partner in either the sample or the carrier. 3. A method for the determination of a substance present in a sample, which comprises: (a) contacting said sample containing a substance having binding sites thereon capable of binding to at least two different immunological binding partners therefor with a mixture of: (i) a first immunological binding partner to said substance, said first immunological binding partner being bound to biotin or a biotin binding protein; and (ii) a detectably labeled second immunological binding partner to said substance; wherein said first immunological binding partner is a monoclonal antibody and said second immunological binding partner is a region specific polyclonal antibody prepared by the method of claim 1; (b) incubating the components of step (a) under conditions sufficient to promote formation of an immune complex between said substance, said first immunological binding partner, and said second immunological binding partner; (c) contacting said complex of step (b) with; (iii) a carrier bound to a biotin binding protein or biotin under conditions sufficient to essentially irreversibly bind said immune complex to said carrier; (d) separating said carrier from the sample; and (e) determining the detactably labeled second immunological binding partner in either the sample or the carrier. 4. The method of claim 2 or 3, wherein said substance is exogenous to the source of said sample. 5. The method of claim 4, wherein said exogenous substance is a pathogen derived antigen. 6. The method of claim 5, wherein said pathogen derived antigen is of viral, bacterial, or parasitic origin. 7. The method of claim 2 or 3, wherein said substance is endogenous to the source of said sample. 8. The method of claim 7, wherein said endogenous substance is a hormone. 9. The method of claim 8, wherein said hormone is selected from the group consisting of human choriogonadotropin, luteinizing hormone, prolactin, follicle stimulating hormone, human growth hormone, somatomedin, thyroid stimulating hormone, parathyroid hormone, adrenocorticotropic hormone, vitamin D, and calcitonin. 10. The method of claim 7, wherein said endogenous substance is a lipid. 11. The method of claim 11, wherein said lipid is selected from the group consisting of cholesterol, lecithin, caroteine, sphingomyelin, cerebroside, and ganglioside. 12. The method of claim 7, wherein said endogenous substance is an enzyme. 13. The method of claim 12, wherein said enzyme is selected from the group consisting of urease, deoxyribonuclease, ribonuclease, creatinine phosphokinase, lactic dehydrogenase, glutamic oxaloacetic transaminase, alkaline phosphatase, 5'-nucleotidase, aspartate aminotransferase, alanine aminotransaminase, and gamma-glutamyl transpeptidase. 14. The method of claim 3, wherein said endogenous substance is an immunoglobulin. 15. The method of claim 2 or 3, wherein said detectable label is an enzyme. 16. The method of claim 15, wherein said enzyme is selected from the group consisting of maleate dehydrogenase, staphylococcal nuclease, delta-5-steroidisomerase, yeast alcohol dehydrogenase, alpha-glycerol-phosphatedehydrogenase, triosephosphateisomerase, horseradish peroxidase, alkaline phosphatase, asparaginase, glucose-oxidase, beta-galactosidase, ribonuclease, urease, catalase, glucose-6-phosphate dehydrogenase, glucoamylase, and acetylcholinesterase. 17. The method of claim 2 or 3 wherein said detectable label is a radioactive isotope. 18. The method of claim 17, wherein said isotope is selected from the group consisting of .sup.3 H, .sup.125 I, .sup.131 I, .sup.32 P, .sup.35 S, .sup.14 C, .sup.51 Cr, .sup.36 Cl, .sup.57 Co, .sup.58 Co, .sup.59 Fe and .sup.75 Se. 19. The method of claim 2 or 3, wherein said detectable label is a fluorescent compound. 20. The method of claim 19, wherein said fluorescent compound is selected from the group consisting of fluorescein isothiocyanate, rhodamine, phycoerythrine, phycocyanine, alophycocyanine, o-phthaldehyde, and fluorescamine. 21. The method of claim 2 or 3, wherein said detectable label is a chemiluminescent compound. 22. The method of claim 21, wherein said chemiluminescent compound is selected from the group consisting of luminol, isoluminol, aromatic acridinium ester, imidazole, acridinium salt, and oxalic ester. 23. The method of claim 2 or 3, wherein said detectable label is a bioluminescent compound. 24. The method of claim 2 or 3, wherein said bioluminescent compound is selected from the group consisting of luciferin and aequorin. 25. The method of claim 2 or 3, wherein said substance is prolactin or human growth hormone. 26. A kit useful for the detection of a substance in a sample comprising carrier means being compartmentalized to receive in close confinement therein one or more containers wherein: (a) a first container contains an immunological binding partner to said substance, said partner bound to biotin or a biotin-binding protein; (b) a second container contains an immunological binding partner to said substance, said partner bound to a detectable label; wherein said first immunological binding partner is a monoclonal antibody and said second immunological binding partner is a region specific polyclonal antibody prepared by the method of claim 11; and (c) a third container contains a solid phase to which a biotin-binding protein or biotin is bound. 27. The kit of claim 26, wherein said substance is an immunoglobulin. 28. The kit of claim 27, wherein said substance is IgE. 29. A kit useful for the detection of a substance in a sample comprising carrier means being compartmentalized to receive in close confinement therein one or more containers wherein: (a) a first container contains a water-soluble mixture of: (i) a first immunological binding partner to said substance, said partner bound to biotin or a biotin-binding protein; and (ii) a second, detectably labeled, immunological binding partner to said substance; wherein said first immunological binding partner is a monoclonal antibody and said second immunological binding partner is a region specific polyclonal antibody prepared by the method of claim 21, and (b) a second container contains a solid phase to which a biotin-binding protein or biotin is bound. 30. The kit of claim 29, wherein said first container (a) contains a mixture of biotin-bound monoclonal antibody to IgE, and region specific polyclonal antibody bound to an enzyme; and said second container (b) contains avidin-coated polystyrene beads. 31. An aqueous soluble composition comprising: a first immunological binding partner to a substance, said substance having binding sites thereon capable of binding to at least two different immunological binding partners, said first binding partner bound to biotin or a biotin-binding protein; and a second, detactably labeled, immunological binding partner to said substance; wherein said first immunological binding partner is a monoclonal antibody and said second immunological binding partner is a region specific polyclonal antibody prepared by the method of claim 1. 32. The composition of claim 3 which is in the form of a homogeneous aqueous solution. 33. A region specific polyclonal antibody to a substance prepared by the method of claim 1. 34. The region specific polyclonal antibody of claim 33 in detactably labeled form.

Details for Patent 4,935,339

Applicant	Tradename	Biologic Ingredient	Dosage Form	BLA	Approval Date	Patent No.	Expiredate
Recordati Rare Diseases, Inc.	ELSPAR	asparaginase	For Injection	101063	01/10/1978		2039-03-29
Nps Pharmaceuticals, Inc.	NATPARA	parathyroid hormone	For Injection	125511	01/23/2015		2039-03-29