title: Sexing murine embryos with an indirect immunofluorescence assay using phage antibody B9-Fab against SDM antigen.
abstract: The use of serologically detectable male (SDM; also called H-Y) antigens to identify male embryos may be limited by the source of anti-SDM antibody. In the present study, novel anti-SDM B9-Fab recombinant clones (obtained by chain shuffling of an A8 original clone) were used to detect SDM antigens on murine embryos. Murine morulae and blastocysts (n=138) were flushed from the oviducts of Kunming mice and incubated with anti-SDM B9-Fab for 30 min at 37°C. With an indirect immunofluorescence assay, the membrane and inner cell mass had bright green fluorescence (presumptive males). Overall, 43.5% (60/138) were classified as presumptive males and 56.5% (78/138) as presumptive females, with 85.0 and 88.5% of these, respectively, confirmed as correct predictions (based on PCR analysis of a male-specific [Sry] sequence). We concluded that the anti-SDM B9-Fab molecule had potential for non-invasive, technically simple immunological sexing of mammalian embryos.
authors.affiliation: College of Veterinary Medicine, Hunan Agricultural University, Changsha, Hunan Province 410128, PR China.
authors.forename: Naidong
authors.initials: N
authors.lastname: Wang
authors.forename: Anwen
authors.initials: A
authors.lastname: Yuan
authors.forename: Jun
authors.initials: J
authors.lastname: Ma
authors.forename: Zhibang
authors.initials: Z
authors.lastname: Deng
authors.forename: Liqun
authors.initials: L
authors.lastname: Xue
mesh_terms: D000818:Animals; D004622:Embryo, Mammalian / Q000276:immunology / Q000502:physiology*; D005260:Female; D019084:Fluorescent Antibody Technique, Indirect / Q000379:methods*; D006182:H-Y Antigen / Q000276:immunology*; D007140:Immunoglobulin Fab Fragments / Q000276:immunology; D008297:Male; D051379:Mice; D011237:Predictive Value of Tests; D012732:Sex Determination Analysis / Q000379:methods*
chemical_list: D006182:H-Y Antigen; D007140:Immunoglobulin Fab Fragments