Polyreactivity of monoclonal antibodies produced against thyroid stimulating hormone (hTSH).
Monoclonal antibodies against human thyroid stimulating hormone (hTSH) are among the key reagents required in the development of an immunoradiometric assay (IRMA) for hTSH in serum. In this study we have produced and characterized twelve hTSH monoclonal antibodies. Hybridomas were generated by fusion of B-lymphocytes from mice immunized with hTSH and myeloma cells. Clones producing antibodies against hTSH were selected using ELISA (Enzyme Linked Immunosorbent Assay). Antibodies from the selected and cloned hybridoma cells were purified by affinity chromatography and their reactivities were tested by ELISA against a panel of antigens, i.e., hTSH, bovine serum albumin (BSA), and milk protein, and also by studying the binding of these monoclonal antibodies with the radioiodinated antigens (hTSH, BSA, and milk protein). It was observed that some of the hTSH monoclonal antibodies produced were polyreactive, reacting with hTSH as well as with unrelated antigen BSA, while others were monoreactive, reacting only to hTSH.
Mirapurkar
S
S
Immunoassay, Labelled Compounds & Jonaki, Board of Radiation and Isotope Technology, Department of Atomic Energy (DAE), Navi Mumbai, India.
Nagvekar
U H
UH
Sivaprasad
N
N
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology* / Q000302:isolation & purification*; D003429:Cross Reactions; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D006825:Hybridomas; D007457:Iodine Radioisotopes / Q000737:chemistry; D051379:Mice; D008894:Milk Proteins / Q000737:chemistry / Q000276:immunology; D012710:Serum Albumin, Bovine / Q000737:chemistry / Q000276:immunology; D013972:Thyrotropin / Q000097:blood / Q000276:immunology*
D000911:Antibodies, Monoclonal; D007457:Iodine Radioisotopes; D008894:Milk Proteins; D012710:Serum Albumin, Bovine; D013972:Thyrotropin