초록 ▼

A method for the detection and/or determination of an antigen having at least two separate antibody combining sites or antigenic determinants (i.e. a polyvalent antigen) using at least two different monoclonal antibodies which bind to the separate sites is described. In particular the method utilize

A method for the detection and/or determination of an antigen having at least two separate antibody combining sites or antigenic determinants (i.e. a polyvalent antigen) using at least two different monoclonal antibodies which bind to the separate sites is described. In particular the method utilizes Protein A with a carrier to immobilize a first monoclonal antibody which in turn, can bind to one antigenic determinant on the antigen. The second antibody with a label is provided in a solution and binds to the second antigenic determinant on the antigen. A test kit is described for practicing the method. Novel anti-PGH synthase and anti-PGI2 synthase antibodies and hybridoma cells producing such antibodies are described.

대표청구항 ▼

In a method for determination of polyvalent antigens by reaction with monoclonal antibodies specific for the antigen, wherein the antigen has at least two separate different combining sites for the antibodies the improvement which comprises: (a) providing a first murine IgG monoclonal antibody which

In a method for determination of polyvalent antigens by reaction with monoclonal antibodies specific for the antigen, wherein the antigen has at least two separate different combining sites for the antibodies the improvement which comprises: (a) providing a first murine IgG monoclonal antibody which combines with a first antigen combining site and which is bound to an immobilizing carrier by means of Staphylococcus aureus Protein A directly or indirectly by means of a conjugal antibody between the IgG antibody and the protein A and a solution containing a second murine IgG monoclonal antibody which combines with a second combining site on the antigen and not to the carrier and which is labeled so as to be detectable, wherein the antibodies are produced by different hybridoma cells and are specific for the same polyvalent antigen but having different combining sites on the antigen, and wherein the second antibody does not react with Protein A or reacts only slowly with Protein A so as not to interfere with determination of the antigen; (b) reacting the antigen in a solution with the first antibody bound to the carrier to provide a first antibody-antigen complex with the carrier; (c) reacting the first complex with the second antibody in a solution to provide a first and second antibody-antigen complex with the carrier which is separable from the solution; (d) separating the first antibody-second antibody-antigen complex from unbound second antibody in solution; and (e) determining the presence of label in the first antibody-second antibody-antigen complex. Monoclonal antibodies produced by hybridoma cells cyo-1,-3,-5 and -7 and deposited as ATCC HB 8124, HB8125, HB8126 and HB 8127 respectively. Monoclonal antibodies produced by hybridoma cells isn-1 and isn-3 and deposited as ATCC HB8123 and HB8122 respectively.