Delayed solid phase immunologic assay
First Claim
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.
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Accused Products
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.
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Citations
34 Claims
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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:
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(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. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 29, 30, 31, 32, 33, 34)
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- 11. The method of claim 11, wherein said lipid is selected from the group consisting of cholesterol, lecithin, caroteine, sphingomyelin, cerebroside, and ganglioside.
Specification