Efficient construction of a large nonimmune phage antibody library: The production of high-affinity human single-chain antibodies to protein antigens
Efficient construction of a large nonimmune phage antibody library: The production of high-affinity human single-chain antibodies to protein antigens
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DOI：
10.1073/pnas.95.11.6157
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发表时间：
1998-05-26
影响因子：
11.1
通讯作者：
Marks, JD
中科院分区：
文献类型：
--
作者：
Sheets, MD;Amersdorfer, P;Marks, JD
通过改进文库构建步骤，产生了含有6.7 × 10（9）个成员的噬菌体展示的人单链抗体（scFv）的大文库。使用14种不同的蛋白质抗原从该文库中亲和选择抗体。用每种抗原分离一组特异性抗体，并且每组含有平均8.7种不同的scFv。抗体-抗原相互作用的测量揭示了低于InM的几种亲和力，与在二次鼠免疫应答期间观察到的亲和力相当。特别地，识别ErbB 2蛋白的四种不同的scFv具有220 pM至4 nM的亲和力。从该文库衍生的抗体被证明是用于免疫测定的有用试剂，例如，产生的针对沙眼衣原体原体的抗体染色衣原体感染的细胞，但不染色未感染的细胞。这些结果证明噬菌体抗体文库理想地适合于快速产生针对多种蛋白质抗原的高亲和力mAb组，这样的文库应该证明对于产生试剂以研究由基因组计划鉴定的基因产物的功能特别有用。
A large library of phage-displayed human single-chain Fv antibodies (scFv), containing 6.7 x 10(9) members, was generated by improving the steps of library construction. Fourteen different protein antigens were used to affinity select antibodies from this library. A panel of specific antibodies was isolated with each antigen, and each panel contained an average of 8.7 different scFv, Measurements of antibody-antigen interactions revealed several affinities below 1 nM, comparable to affinities observed during the secondary murine immune response, In particular, four different scFv recognizing the ErbB2 protein had affinities ranging from 220 pM to 4 nM. Antibodies derived from the library proved to be useful reagents for immunoassays, For example, antibodies generated to the Chlamydia trackomatis elementary bodies stained Chlamydia-infected cells, but not uninfected cells, These results demonstrate that phage antibody libraries are ideally suited for the rapid production of panels of high-affinity mAbs to a wide variety of protein antigens, Such libraries should prove especially useful for generating reagents to study the function of gene products identified by genome projects.