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Monoclonal antibodies against zona pellucida
54
M O N O C L O N A L A N T I B O D I E S AGAINST ZONA P E L L U C I D A
A.K. BAMEZAI, A.H. BAND, A . B H A T T A C H A R Y A , R.D. GAUR, S. SEHGAL, C. DAS AND G.P. TALWAR DEPARTMENT OF BIOCHEMISTRY,AIIMS & NIl, Nh-W DELHI-II0029, INDIA.
SUMAN
M o n o c l o n a l antibodies were raised against porcine zona pellucida by fusing mouse splenocytes with Sp2/o Agl4myeloma. Clones were screened by a solid phase enzyme immunoassay. Six stable lines were established. Five of them were IgGl, k and one was IgG2, k. Antibodies of all the six clones were n o n - r e a c t i v e with erythrocytes, liver and other tissues but reacted strongly with zona. The clones were grown as ascites fluid in Balb/c mice. The ascites at dilution of 103 inhibited the lysis of zona by trypsin for as long as 24 hrs of o b s e r v a t i o n period. The antibodies were also able to p a r t i a l l y inhibit the binding of spermatozoa to zona in an in vitro assay. The effect of antibodies given p a s s i v e l y to m o ~ e y s on fertility and hormonal profiles are under study. i~ONOCLONAL ANTIBOUIES TESTED SY ST E~,i
University
IN THE ZONA FREE HAhSTER OOCYTE
H.-H. Riedel, V.Baukloh, T . W a h e - R o h r b a c h and L.[.ettler, Department of Obstetrics and Gynecology, of Kiel ,F'.egewischstr. 4, 2300 Kiel, FRG
The influence of some monoclooal antibodies on the penetration capacity of human spermatozoa was investigateP in the zona Free hamster oocyte system. Attempts were made to influence penetration rates in this system by additing culture supernatants and mice ascites containing different types of monoclonal antibodies to the incubation medium. The method used for the generation of hybridoma was that reported by K~hler and Trilstein in 1878. The crossed reactivity test of the monoclonal sperm antibodies with other human tissue revealed 32 hybridoma cultures out of the 149 generated, that produced antibody not reactive with other tissues such as tonsil, cardiac muscle,ovary, kidney etc. 5 of these hybridoma cultures secreted antibodies effecting sperm a g g l u t i n a t i o n and 5 caused sperm immobilization. Within the zona free hamster oocyte system especially the monoclonal antibodies of type III 3-I, I 18-1 and VII 5 were tested with ejaculates of 3 donors of proven fertility. All the monoclonal antiboOies were tested in at least 3 parallels with the 3 donor ejaculates. The observed reduction in penetration rates varied between 21 and 3 5 ~ Now we test some other monoclonal antibodies. The final results will be presented in Kyoto. Characterization of human serum factors bound to zona pellucida of porcine oocytes Minoru Shigeta, Morihiro Takeda, Kqj~ Koyama and Shinzo Isojima Dept. Obst. & Gyn., Hyogo Medical College, Nishinomiya, Japan Human and porcine zonae pellucidae (ZP) have cross-reactive antigens and
M O N O C L O N A L A N T I B O D I E S AGAINST ZONA P E L L U C I D A
A.K. BAMEZAI, A.H. BAND, A . B H A T T A C H A R Y A , R.D. GAUR, S. SEHGAL, C. DAS AND G.P. TALWAR DEPARTMENT OF BIOCHEMISTRY,AIIMS & NIl, Nh-W DELHI-II0029, INDIA.
SUMAN
M o n o c l o n a l antibodies were raised against porcine zona pellucida by fusing mouse splenocytes with Sp2/o Agl4myeloma. Clones were screened by a solid phase enzyme immunoassay. Six stable lines were established. Five of them were IgGl, k and one was IgG2, k. Antibodies of all the six clones were n o n - r e a c t i v e with erythrocytes, liver and other tissues but reacted strongly with zona. The clones were grown as ascites fluid in Balb/c mice. The ascites at dilution of 103 inhibited the lysis of zona by trypsin for as long as 24 hrs of o b s e r v a t i o n period. The antibodies were also able to p a r t i a l l y inhibit the binding of spermatozoa to zona in an in vitro assay. The effect of antibodies given p a s s i v e l y to m o ~ e y s on fertility and hormonal profiles are under study. i~ONOCLONAL ANTIBOUIES TESTED SY ST E~,i
University
IN THE ZONA FREE HAhSTER OOCYTE
H.-H. Riedel, V.Baukloh, T . W a h e - R o h r b a c h and L.[.ettler, Department of Obstetrics and Gynecology, of Kiel ,F'.egewischstr. 4, 2300 Kiel, FRG
The influence of some monoclooal antibodies on the penetration capacity of human spermatozoa was investigateP in the zona Free hamster oocyte system. Attempts were made to influence penetration rates in this system by additing culture supernatants and mice ascites containing different types of monoclonal antibodies to the incubation medium. The method used for the generation of hybridoma was that reported by K~hler and Trilstein in 1878. The crossed reactivity test of the monoclonal sperm antibodies with other human tissue revealed 32 hybridoma cultures out of the 149 generated, that produced antibody not reactive with other tissues such as tonsil, cardiac muscle,ovary, kidney etc. 5 of these hybridoma cultures secreted antibodies effecting sperm a g g l u t i n a t i o n and 5 caused sperm immobilization. Within the zona free hamster oocyte system especially the monoclonal antibodies of type III 3-I, I 18-1 and VII 5 were tested with ejaculates of 3 donors of proven fertility. All the monoclonal antiboOies were tested in at least 3 parallels with the 3 donor ejaculates. The observed reduction in penetration rates varied between 21 and 3 5 ~ Now we test some other monoclonal antibodies. The final results will be presented in Kyoto. Characterization of human serum factors bound to zona pellucida of porcine oocytes Minoru Shigeta, Morihiro Takeda, Kqj~ Koyama and Shinzo Isojima Dept. Obst. & Gyn., Hyogo Medical College, Nishinomiya, Japan Human and porcine zonae pellucidae (ZP) have cross-reactive antigens and