Rapid and efficient immobilization of soluble and small particulate antigens for solid phase radioimmunoassays.
A rapid method of antigen immobilization (10 min.) was developed using soluble antigens (bovine serum albumin, epidermal growth factor, and goat IgG) and small particulate antigens (Keyhole limpet hemocynanine and E. coli) by drying them on filter paper discs. This technique results in a high % of the soluble antigen remaining firmly bound, goat IgG (89%), bovine serum albumin (73%). All the antigens we tested retained their antigenicity after drying as detected by [125I] labeled staphylococcal protein A radioimmunoassay. This method of antigen immobilization was compared to adsorption to plastic wells and was found to be much faster (10 min vs 18 hrs) and was 5 times more efficient than adsorption to plastic wells. Using this technique, we were able to detect as little as 40 ng of bovine serum albumin. These characteristics suggest that this technique of soluble antigen immobilization may be useful in rapid detection of antibodies to many different antigens, as well as detecting ng amounts of the antigens directly.
D000818:Animals; D000906:Antibodies; D000941:Antigens; D002417:Cattle; D005374:Filtration / Q000295:instrumentation; D011817:Rabbits; D011863:Radioimmunoassay / Q000379:methods*; D012710:Serum Albumin, Bovine; D012995:Solubility
D000906:Antibodies; D000941:Antigens; D012710:Serum Albumin, Bovine