Nishinaka S, Akiba H, Nakamura M, Suzuki K, Suzuki T, Tsubokura K, Horiuchi H, Furusawa S, Matsuda H
Department of Bio-Development, Applied Technology Research Center, NKK Corporation, Kawasaki, Japan.
J Vet Med Sci. 1996 Nov;58(11):1053-6. doi: 10.1292/jvms.58.11_1053.
Two chicken B cell lines, termed MuH1 and MuH4, which are resistant to ouabain, were established as fusion partners for production of chicken monoclonal antibody from the parental cell lines R27H1 and R27H4 which were deficient in thymidine kinase activity. The MuH1 synthesized, but did not secrete mu chain, whereas the MuH4 secreted non-specific IgM. Fusions of the MuH1 and MuH4 with spleen cells from chickens immunized with human IgG resulted in successful preparations of hybridomas secreting antibodies specific to human IgG. Transformed cells insensitive to HAT appeared in many culture wells after cell hybridizations with R27H4, but on hybridization with MuH1 or MuH4 the appearance of such unfavorable cells could be avoided by the inclusion of ouabain in HAT medium. Thus, in comparison with the R27H4 line, the MuH1 and MuH4 lines did not provide higher fusion efficiencies, but gave increased opportunities to obtain hybridomas producing specific antibodies. Among a total of eight hybridomas, four derived from HuM1 and four derived from MuH4, seven secreted both IgG and IgM, and the remaining one derived from MuH1 secreted only IgG. In all eight hybridoma lines IgG was specifically reactive to human IgG.
建立了两种对哇巴因具有抗性的鸡B细胞系，分别称为MuH1和MuH4，作为从缺乏胸苷激酶活性的亲本细胞系R27H1和R27H4生产鸡单克隆抗体的融合伙伴。MuH1合成但不分泌μ链，而MuH4分泌非特异性IgM。用人类IgG免疫的鸡的脾细胞与MuH1和MuH4融合，成功制备了分泌对人类IgG具有特异性的抗体的杂交瘤。与R27H4进行细胞杂交后，许多培养孔中出现了对HAT不敏感的转化细胞，但与MuH1或MuH4杂交时，通过在HAT培养基中加入哇巴因可以避免此类不良细胞的出现。因此，与R27H4细胞系相比，MuH1和MuH4细胞系并没有提供更高的融合效率，但增加了获得产生特异性抗体的杂交瘤的机会。在总共8个杂交瘤中，4个源自HuM1，4个源自MuH4，其中7个同时分泌IgG和IgM，其余1个源自MuH1，仅分泌IgG。在所有8个杂交瘤系中，IgG对人类IgG具有特异性反应。
