Poul M A, Cerutti M, Chaabihi H, Ticchioni M, Deramoudt F X, Bernard A, Devauchelle G, Kaczorek M, Lefranc M P
Laboratoire d'ImmunoGénétique Moléculaire, CNRS, Universités Montpellier I et II, France.
Eur J Immunol. 1995 Jul;25(7):2005-9. doi: 10.1002/eji.1830250731.
Plasmid cassette-transfer vectors pBHuC chi and pBHuC gamma l have been designed which enable the construction of recombinant baculoviruses directing the co-expression of complete immunoglobulin in insect cells. We describe the application of these vectors for the expression of a human/mouse chimeric monoclonal antibody of potential immunosuppressive clinical value derived from a mouse anti-human CD29 monoclonal antibody (Mu-K20). The chimeric K20 light and heavy chains produced in sf9 insect cells were correctly processed and assembled into a normal immunoglobulin which is secreted into the culture medium of infected cells. The chimeric mAb Ch-K20-sf9 reproduces in vitro the functional properties of the parental mouse K20, including affinity and inhibition of lymphocyte proliferation. These results demonstrate that the baculovirus/insect cell expression system is suitable for the expression of fully active monoclonal antibodies of therapeutic value. Our generic cassette approach makes this system a very flexible and convenient one for the rapid production of either chimeric, humanized or human mAb with heavy and light chains of any isotype.
已设计出质粒盒式转移载体pBHuC chi和pBHuC gamma l,它们能够构建在昆虫细胞中指导完整免疫球蛋白共表达的重组杆状病毒。我们描述了这些载体在表达源自小鼠抗人CD29单克隆抗体(Mu-K20)的具有潜在免疫抑制临床价值的人/鼠嵌合单克隆抗体中的应用。在sf9昆虫细胞中产生的嵌合K20轻链和重链经过正确加工,并组装成正常的免疫球蛋白,分泌到感染细胞的培养基中。嵌合单克隆抗体Ch-K20-sf9在体外重现了亲本小鼠K20的功能特性,包括亲和力和对淋巴细胞增殖的抑制作用。这些结果表明,杆状病毒/昆虫细胞表达系统适用于表达具有治疗价值的完全活性单克隆抗体。我们通用的盒式方法使该系统成为一种非常灵活和方便的系统,可用于快速生产具有任何同种型重链和轻链的嵌合、人源化或人单克隆抗体。