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H-Y 에 대한 단일클론 항체의 생산 및 그 이용에 관한 연구 1 . H-Y 에 대한 단일클론항체의 생산
심호섭(H . S . Shim),김재화(J . H . Kim),이병철(B . C . Lee),김종배(J . B . Kim),박홍양(H . Y . Park),정길생(K . S . Chung) 한국축산학회 1988 한국축산학회지 Vol.30 No.7
Testis supernatant, a source of H-Y, obtained from BALB/c mice was used to immunize females of same strain. B lymphocytes of mouse producing antibodies to H-Y were fused with SP2/0-Ag 14 myeloma cells and distributed to 384 wells of 96-well microtiter plates. Eighty hybridoma colonies were formed, resulting in 20.8 percent of fusion efficiency. Three strong positive wells from hybridoma colonies were selected for cloning by ELISA and two of them were also found to be positive by indirect immunofluorescence test. Twelve wells of ELISA-positive were selected after cloning and 2D45D4 clones from them were confirmed to produce monoclonal antibodies to H-Y by indirect immunofluorescence test.
생쥐수정란에 대한 H - Y 항체처리가 산자의 성비에 미치는 영향
심호섭,고정재,김종배,박홍양,정길생 ( H . S . Shim,J . J . Ko,J . B . Kim,H . Y . Park,K . S . Chung ) 한국축산학회 1986 한국축산학회지 Vol.28 No.12
These experiments were carried out to control the sex of offsprings in mice by sexing embryos using immunological means prior to transfer to pseudopregnant recipients. H-Y antisera were prepared in inbred SD female rats by repeated immunization of testis supernatant and spleen cells from males of same strain. ELISA and indirect immunofluorescence test were used to detect H-Y antibody in antisera. Eight- to 16-cell mouse embryos were cultured in medium with H-Y antibody and complement (treated embryos) and in medium with BSA (control embryos). After 24-48 hr of culture, embryos were observed their morphological characteristics under the phase contrast microscope. Embryos developed to normal blastocyst were transferred to pseudopregnant recipients and sex of resultant offspring was investigated. The results obtained in these experiments were summarized as follows: 1. Production of H-Y antibodies in antisera obtained from immunized rats was confimed by ELISA and indirect immunofluorescence test. 2. Of 270 embryos treated with H-Y antibody and complement, 126 embryos (46.7%) were developed to normal blastocysts. 3. Following transfer of 126 blastocysts, 16 embryos (12.6%) were survived to term and 13 females (81.3%), significantly high ratio of female offspring, were produced.