ELISA solid phase: partial denaturation of coating antibody yields a more efficient solid phase.
We have examined conditions which will improve the solid phase in ELISAs for HBsAg and ferritin in human serum. Sheep and rabbit antibodies were used. It was found that pre-exposure of antibody to low pH, 3 M urea and temperatures as high as 82 degrees C will enhance ELISA colour in these assays. Further experiments indicate that these conditions perturb the structure of the antibody molecules and results in the exposure of new hydrophobic regions. It is speculated that these more hydrophobic molecules can bind to regions on the plastic surface normally not coated by non-perturbed molecules.
D000478:Alkylation; D000818:Animals; D000906:Antibodies / Q000032:analysis*; D000914:Antibodies, Viral / Q000032:analysis*; D002942:Circular Dichroism; D004797:Enzyme-Linked Immunosorbent Assay; D005293:Ferritins / Q000276:immunology; D006510:Hepatitis B Antibodies / Q000032:analysis*; D006514:Hepatitis B Surface Antigens / Q000276:immunology; D006358:Hot Temperature; D006801:Humans; D006863:Hydrogen-Ion Concentration; D011489:Protein Denaturation; D011817:Rabbits; D012756:Sheep; D014508:Urea / Q000494:pharmacology
D000906:Antibodies; D000914:Antibodies, Viral; D006510:Hepatitis B Antibodies; D006514:Hepatitis B Surface Antigens; D014508:Urea; D005293:Ferritins