Single amino acid substitution in the mouse IgG1 Fc region induces drastic enhancement of the affinity to protein A
Single amino acid substitution in the mouse IgG1 Fc region induces drastic enhancement of the affinity to protein A
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DOI：
10.1093/proeng/gzg037
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发表时间：
2003-04-01
期刊：
影响因子：
--
通讯作者：
Akaike, T
中科院分区：
文献类型：
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作者：
Nagaoka, M;Akaike, T
单克隆抗体的纯化有时需要大量时间，并且涉及复杂的步骤，因为小鼠IgG与蛋白A或蛋白G相互作用的能力比来自其他物种如人和兔的IgG差。为了解决这个问题，我们将小鼠IgG Fc区的一个或两个氨基酸残基与人IgG的Fc区的氨基酸残基交换。三个突变体（T252 M、T254 S和T252 M-T254 S）显示出对蛋白A的亲和力的显著改善。将苏氨酸252残基交换为甲硫氨酸（T252 M）是最有效的。该结果表明，直接和简单的修饰允许有效纯化单克隆抗体和含有小鼠IgG Fc区的融合蛋白。
The purification of monoclonal antibody sometimes requires a lot of time and involves complicated steps because of the poorer ability of mouse IgG to interact with protein A, or also with protein G, than IgGs from other species such as those of human and rabbit. To resolve this problem, we exchanged one or two amino acid residues of mouse IgG Fc region with that of human IgG. Three mutants (T252M, T254S and T252M-T254S) showed significant improvement in the affinity to protein A. The exchange of the threonine 252 residue to methionine (T252M) was most efficient. This result suggests that a direct and simple modification allows the efficient purification of monoclonal antibody and of fusion protein containing mouse IgG Fc region.