Barnard K, James M P
Biosci Rep. 1984 Mar;4(3):195-201. doi: 10.1007/BF01119654.
An enzyme immunoassay, using finely ground rabbit glomerular basement membrane (GBM) as an antigen, was able to detect sheep anti-rabbit GBM antibodies at serum dilutions of 1:32 000. The particulate GBM bound firmly to plastic micro ELISA plates without the aid of a linking agent, and the antigen-coated plates remained stable for several months when stored at -70 degrees C. There were no appreciable differences between amino acid compositions of ground and whole GBM, and no detectable loss of antigens occurred during the grinding procedure. Competitive inhibition assays with collagenase and pepsin digests of rabbit GBM demonstrated preservation of collagenous and non-collagenous antigens in the ground GBM. The assay should prove to be a relatively simple and highly sensitive technique for detecting antibodies to a wide spectrum of GBM antigens.
一种酶免疫测定法，使用精细研磨的兔肾小球基底膜（GBM）作为抗原，能够在血清稀释度为1:32000时检测到羊抗兔GBM抗体。颗粒状GBM在不借助连接剂的情况下牢固地结合到塑料微量酶联免疫吸附测定板上，并且抗原包被的板在-70℃储存时可保持稳定数月。研磨后的GBM与完整GBM的氨基酸组成没有明显差异，并且在研磨过程中未检测到抗原损失。用兔GBM的胶原酶和胃蛋白酶消化物进行的竞争性抑制试验表明，研磨后的GBM中胶原性和非胶原性抗原得以保留。该测定法应被证明是一种相对简单且高度灵敏的技术，可用于检测针对多种GBM抗原的抗体。
