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Insolubilized proteins and immunoassays utilizing them
| Details |
Inventors: Limet, Joseph; Cambiaso, Cesar L.; Moussebois, Claude H.; Masson, Pierre L.;
Assignee: Technicon Instruments Corporation (Tarrytown, NY)
Primary Examiner: Marantz; Sidney
Assistant Examiner:
Attorney, Agent or Firm: Tedesco; S. P.
Insolublized antibodies, F(ab').sub.2 fragments thereof and protein antigens are made by coupling to a water-insoluble substrate using a bridging agent which covalently binds to sulphur atoms in antibody, fragment or protein rather than to free amino groups therein. This is preferably effected by using a chloracetyl-terminated bridging agent. Immunoassays utilizing these insolubilized reagents are described. One novel class of bridging agents comprising anhydride and monochloracetyl end groups, which is useful for making the said reagents, is also disclosed. |
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DETAILED DESCRIPTION We claim: 1.
A reagent for use in immunoassays which comprise a protein antigen, an antibody or the F(ab').
sub.
2 fragments of an antibody, covalently bonded to a water-in-soluble substrate by a bridging group which is directly linked to sulphur atoms in the said antigen, antibody or fragments, that portion of the bridging group linked directly to the said sulphur atoms being derived from a monochloracetyl group.
2.
A reagent according to claim 1 wherein the bridging group contains a chain of at least 5 atoms length to separate spatially the said antigen, antibody or fragment, from the substrate.
3.
A reagent according to claim 1, wherein the said antibody is an immunoglobulin G.
4.
A reagent according to claim 1, which comprises the F(ab').
sub.
2 fragments of an immunoglobulin G, and which is substantially free from the said immunoglobulin G and F(c) fragments thereof.
5.
A reagent according to claim 1, wherein the substrate comprises a protein to which the said bridging group is directly linked, and wherein that portion of the bridging group which is linked to the protein of the substrate is derived from an anhydride group.
6.
A reagent according to claim 5, wherein the bridging group is derived from a compound of the formula ##STR5## wherein n is an integer from 1 to 10.
7.
A reagent according to claim 1, wherein the substrate is in sheet-like form or in the form of a hollow tube, or is a particulate material.
8.
A reagent according to claim 7, wherein the substrate is a latex, or is a particulate material which is magnetically attractable.
9.
A reagent for use in immunoassays which comprises a water-insoluble particulate material in suspension in a fluid, the particulate material having a protein coating to which is covalently linked the anhydride group of a bridging agent of the formula ##STR6## wherein n is from 1 to 10, the chloroacetyl group of said agent being directly bonded to sulphur atoms in an antigen protein, an antibody or the F(ab').
sub.
2 fragments of an immunoglobulin G antibody
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