Temperature-dependent non-specific adsorption of recombinant bovine leukemia virus receptor BLVRcp1 in immunoassay.
Recombinant bovine leukemia virus receptor, BLVRcp1, possessed the unusual property of binding plastic plates after blocking nonspecific binding sites. Adhesiveness of BLVRcp1 to blocked plates hindered development of an antigen capture and receptor binding assay with this protein. Unexpectedly, non-specific adsorption of BLVRcp1 was dramatically influenced by temperature. Optimizing incubation temperature and antigen capture at 4 degrees C instead of 37 degrees C and the use of milk as blocking solution removed nonspecific binding of BLVRcp1 allowing development of a functional immunoassay. Thus, the temperature used for antigen capture can be a critical factor that influences performance of the immunoassay.
Orlik
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O
Department of Animal Health and Biomedical Sciences, University of Wisconsin-Madison 53706, USA.
Altaner
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C
Splitter
G A
GA
D016475:3T3 Cells; D000327:Adsorption; D000818:Animals; D001665:Binding Sites / Q000187:drug effects / Q000276:immunology; D001667:Binding, Competitive / Q000276:immunology; D015592:Immunoradiometric Assay / Q000379:methods; D001909:Leukemia Virus, Bovine / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D011485:Protein Binding / Q000187:drug effects / Q000276:immunology; D011991:Receptors, Virus / Q000235:genetics / Q000378:metabolism*; D011994:Recombinant Proteins / Q000378:metabolism*; D012710:Serum Albumin, Bovine / Q000494:pharmacology; D013499:Surface Properties; D013696:Temperature
D011991:Receptors, Virus; D011994:Recombinant Proteins; D012710:Serum Albumin, Bovine