Orlik O, Altaner C, Splitter G A
Department of Animal Health and Biomedical Sciences, University of Wisconsin-Madison 53706, USA.
J Immunoassay. 1997 May;18(2):185-98. doi: 10.1080/01971529708005812.
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.
重组牛白血病病毒受体BLVRcp1在封闭非特异性结合位点后具有结合塑料板的特殊性质。BLVRcp1与封闭板的粘附性阻碍了用该蛋白进行抗原捕获和受体结合测定的开展。出乎意料的是，BLVRcp1的非特异性吸附受温度的显著影响。将孵育温度和抗原捕获温度优化为4℃而非37℃，并使用牛奶作为封闭溶液，消除了BLVRcp1的非特异性结合，从而能够开展功能性免疫测定。因此，用于抗原捕获的温度可能是影响免疫测定性能的关键因素。
