Veerhuis R, Hendriksen P J, Hengst A M, Kruijt L, Tieman M, Booman P
Immunobiology Department, DLO-Research Institute for Animal Production (IVO-DLO) Schoonoord, AM Zeist, Netherlands.
Vet Immunol Immunopathol. 1994 Sep;42(3-4):317-30. doi: 10.1016/0165-2427(94)90076-0.
In an attempt to improve the accuracy of sexing bovine embryos, new anti-H-Y monoclonal antibodies were produced and selected, using an extended screening procedure. In addition to the commonly used screening of soluble H-Y antigen sources, such as testis supernatant and Daudi supernatant, the binding specificity to cell surface H-Y antigen was tested also. A radioimmunoassay (RIA) employing male and, as a control, female bovine lymphocytes, and enzyme-linked immunosorbent assays (ELISAs) on solubilized membrane fractions resulted in the selection of a number of clones producing monoclonal antibody (mAb) with male-enhanced binding. Four of the anti-H-Y mAb were assessed for binding to Day 7 or 8 bovine embryos. The accuracy of sexing bovine embryos ranged from 58% to 71%. Two of the four antibodies did not react with presumed soluble H-Y antigen-containing sources in an ELISA. These results raise doubts about the suitability of the presumed soluble H-Y antigen sources, Daudi, TM4 and testis supernatant, to be used in screening tests for anti-H-Y antibodies.
为提高牛胚胎性别的鉴定准确性，采用了一种扩展的筛选程序来制备和筛选新的抗H-Y单克隆抗体。除了常用的对可溶性H-Y抗原来源（如睾丸上清液和Daudi上清液）进行筛选外，还测试了对细胞表面H-Y抗原的结合特异性。使用雄性牛淋巴细胞作为对照，通过放射免疫测定法（RIA）以及对溶解的膜组分进行酶联免疫吸附测定法（ELISA），筛选出了一些产生与雄性结合增强的单克隆抗体（mAb）的克隆。对4种抗H-Y单克隆抗体与第7天或第8天的牛胚胎的结合情况进行了评估。牛胚胎性别的鉴定准确率在58%至71%之间。这4种抗体中有2种在ELISA中不与推测含有可溶性H-Y抗原的来源发生反应。这些结果使人对推测的可溶性H-Y抗原来源Daudi、TM4和睾丸上清液用于抗H-Y抗体筛选试验的适用性产生怀疑。
