Poul M A, Cerutti M, Chaabihi H, Devauchelle G, Kaczorek M, Lefranc M P
Laboratoire d'ImmunoGénétique Moléculaire, Institut de Génétique Moléculaire, UMR 9942, CNRS, Montpellier, France.
Immunotechnology. 1995 Dec;1(3-4):189-96. doi: 10.1016/1380-2933(95)00019-4.
Various systems have been described for the expression of recombinant monoclonal antibodies for therapeutical applications. Insect cells offer great advantages with respect to post-translational modifications, stability, yields and applications.
To construct plasmid cassette transfer vectors in order to express chimeric, humanized or human antibodies in insect cells using baculovirus expression system.
Two transfer vectors, pBHuC kappa and pBHuC gamma 1, were designed. They contain a viral promoter (polyhedrin or p10 promoters, respectively), a signal peptide sequence and a human immunoglobulin light chain C kappa gene or heavy chain C gamma 1 sequence, respectively. Restriction sites have been introduced to allow insertion of rearranged variable genes, after amplification by polymerase chain reaction.
Recombinant baculoviruses expressing complete immunoglobulins have been generated by a double-recombination event between baculovirus DNA and the loaded cassette transfer vectors.
Our genetic cassette approach makes this system a very flexible and convenient one for the rapid production of therapeutic monoclonal antibodies with heavy and light chains of any human isotype. Specific variable regions selected by the antibody phage display technology can be easily transferred in these vectors to obtain a complete antibody.
已描述了多种用于治疗应用的重组单克隆抗体表达系统。昆虫细胞在翻译后修饰、稳定性、产量及应用方面具有很大优势。
构建质粒盒式转移载体，以便使用杆状病毒表达系统在昆虫细胞中表达嵌合抗体、人源化抗体或人抗体。
设计了两种转移载体，即pBHuC κ和pBHuC γ1。它们分别包含一个病毒启动子（分别为多角体蛋白启动子或p10启动子）、一个信号肽序列以及一个人免疫球蛋白轻链Cκ基因或重链Cγ1序列。已引入限制性酶切位点，以便在通过聚合酶链反应扩增后插入重排的可变基因。
通过杆状病毒DNA与加载的盒式转移载体之间的双重组事件产生了表达完整免疫球蛋白的重组杆状病毒。
我们的基因盒方法使该系统成为一种非常灵活且方便的系统，可快速生产具有任何人同种型重链和轻链的治疗性单克隆抗体。通过抗体噬菌体展示技术选择的特定可变区可轻松转移至这些载体中以获得完整抗体。
