PURIFICATION OF ANTIBODIES USING PROTEIN-L-BINDING FRAMEWORK STRUCTURES IN THE LIGHT-CHAIN VARIABLE DOMAIN
PURIFICATION OF ANTIBODIES USING PROTEIN-L-BINDING FRAMEWORK STRUCTURES IN THE LIGHT-CHAIN VARIABLE DOMAIN
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DOI：
10.1016/0022-1759(93)90273-a
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发表时间：
1993-08-26
影响因子：
2.2
通讯作者：
AKERSTROM, B
中科院分区：
文献类型：
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作者：
NILSON, BHK;LOGDBERG, L;AKERSTROM, B
大胃链球菌（Peptostreptoccus magnus）的蛋白L特异性结合到Ig轻链的可变结构域，而不干扰抗原结合位点。在这项工作中，一个基因工程的蛋白质L片段，包括四个重复的igg结合重复单位，用于从各种来源纯化Ig。因此，利用蛋白L-Sepharose亲和层析法从人和小鼠血清中一次性纯化IgG、IgM和IgA。此外，用蛋白L-Sepharose从杂杂瘤细胞或产生抗体的细菌细胞培养物中纯化人、小鼠单克隆IgG、IgM、IgA和人IgG Fab片段，以及小鼠/人嵌合重组抗体。人源化小鼠抗体也是如此，将小鼠高变抗原结合区引入蛋白l结合的kappa亚型III人IgG中。这些实验表明，用蛋白l结合框架区设计抗体和抗体片段（Fab, Fv）是可能的，然后可以用于基于蛋白l的纯化方案。
Protein L from the bacterial species Peptostreptoccus magnus binds specifically to the variable domain of Ig light chains, without interfering with the antigen-binding site. In this work a genetically engineered fragment of protein L, including four of the repeated Ig-binding repeat units, was employed for the purification of Ig from various sources. Thus, IgG, IgM, and IgA were purified from human and mouse serum in a single step using protein L-Sepharose affinity chromatography. Moreover, human and mouse monoclonal IgG, IgM, and IgA, and human IgG Fab fragments, as well as a mouse/human chimeric recombinant antibody, could be purified from cultures of hybridoma cells or antibody-producing bacterial cells, with protein L-Sepharose. This was also the case with a humanized mouse antibody, in which mouse hypervariable antigen-binding regions had been introduced into a protein L-binding kappa subtype III human IgG. These experiments demonstrate that it is possible to engineer antibodies and antibody fragments (Fab, Fv) with protein L-binding framework regions, which can then be utilized in a protein L-based purification protocol.