Mocikat R, Kardinal C, Lang P, Zeidler R, Thierfelder S
GSF-Institut für Immunologie, Klinische Molekularbiologie, München, Germany.
Immunology. 1995 Jan;84(1):159-63.
Chimeric antibodies against the murine T-cell antigen Thy-1.2 were generated in amounts sufficient for in vivo studies by substituting the constant gene segments via homologous recombination in the hybridoma cell. We show that an integration vector targets the heavy chain locus at high frequency even in a non-isogenic situation. Using this vector type, for the first time expression rates were obtained that were identical to the parental hybridoma. The use of the gpt selection marker seems to be crucial for efficient expression, and may overcome a recently claimed drawback of vector integration. A chimeric antibody produced by gene targeting was characterized in vitro and in vivo.
通过在杂交瘤细胞中进行同源重组替换恒定基因片段，产生了足以用于体内研究的抗小鼠T细胞抗原Thy-1.2嵌合抗体。我们表明，即使在非同基因情况下，整合载体也能高频靶向重链基因座。使用这种载体类型，首次获得了与亲本杂交瘤相同的表达率。gpt选择标记的使用似乎对有效表达至关重要，并且可能克服最近声称的载体整合缺点。对通过基因靶向产生的嵌合抗体进行了体外和体内表征。
