Monoclonal antibody based sandwich enzyme-linked immunosorbent assay for chicken growth hormone.
1. Monoclonal antibodies which bind to different epitopes of chicken growth hormone (cGH) were used to develop a homologous sandwich enzyme-linked immunosorbent assay (ELISA). 2. The first antibody, which is species specific, was immobilised on microtitre plates and concentrations of cGH in biological fluids were estimated by revealing bound hormone using a second, biotinylated monoclonal antibody. 3. The sensitivity was 0.024 ng/ml, which is at least ten-fold greater than current radioimmunoassays (RIA) and there was no cross-reactivity to other chicken pituitary hormones or to growth hormone from other species. 4. The accuracy and precision of the assay were similar to RIA, and the growth hormone concentrations measured in plasma samples by both RIA and this new ELISA showed a high degree of correlation. 5. The assay takes only 4 h using pre-coated plates which can be stored at 4 degrees C in sucrose. The advantages of being rapid and non-isotopic make this method attractive to both research and industrial laboratories.
Houston
B
B
Department of Cellular and Molecular Biology, AFRC Institute of Animal Physiology and Genetics Research, Edinburgh Research Station, Roslin, Midlothian, Scotland.
Peddie
D
D
Goddard
C
C
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000276:immunology*; D000918:Antibody Specificity; D002645:Chickens / Q000097:blood*; D003429:Cross Reactions; D004306:Dose-Response Relationship, Immunologic; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000276:immunology; D005260:Female; D013006:Growth Hormone / Q000032:analysis* / Q000276:immunology; D006825:Hybridomas; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D011237:Predictive Value of Tests; D012044:Regression Analysis; D013045:Species Specificity
D000911:Antibodies, Monoclonal; D000939:Epitopes; D013006:Growth Hormone