The production of anti-H-Y monoclonal antibodies: their potential use in a sex test for bovine embryos.
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.
Veerhuis
R
R
Immunobiology Department, DLO-Research Institute for Animal Production (IVO-DLO) Schoonoord, AM Zeist, Netherlands.
Hendriksen
P J
PJ
Hengst
A M
AM
Kruijt
L
L
Tieman
M
M
Booman
P
P
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis*; D002417:Cattle; D002460:Cell Line; D002462:Cell Membrane / Q000276:immunology; D004622:Embryo, Mammalian / Q000502:physiology*; D004797:Enzyme-Linked Immunosorbent Assay / Q000662:veterinary; D005260:Female; D006182:H-Y Antigen / Q000276:immunology*; D006801:Humans; D007074:Immunoglobulin G / Q000276:immunology; D008214:Lymphocytes / Q000276:immunology; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D011863:Radioimmunoassay / Q000662:veterinary; D012733:Sex Differentiation / Q000276:immunology*; D013737:Testis / Q000276:immunology
D000911:Antibodies, Monoclonal; D006182:H-Y Antigen; D007074:Immunoglobulin G