Single chain variable fragment antibodies selected by phage display against the sporozoite surface antigen S16 of Cryptosporidium parvum
Single chain variable fragment antibodies selected by phage display against the sporozoite surface antigen S16 of Cryptosporidium parvum
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DOI：
10.1016/j.exppara.2010.01.001
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发表时间：
2010-06-01
影响因子：
2.1
通讯作者：
Trevors, Jack T.
中科院分区：
文献类型：
--
作者：
Boulter-Bitzer, Jeanine I.;Lee, Hung;Trevors, Jack T.
使用两个人单链可变片段（scFv）文库来选择与微小隐孢子虫的表面糖蛋白S16结合的克隆。淘选Tomlinson文库I和J导致分离9个不同的克隆。在具有全长scFv的四个克隆中，三个含有终止密码子。其余5个克隆被截短，其中4个缺失重链，1个缺失大部分轻链。全长克隆表现出更好的结合天然C。parvum蛋白和重组S16相比，截短的克隆，除了一个截短的克隆。没有一个选择的克隆与贾第虫、大肠杆菌、化脓性链球菌、单核细胞增生李斯特菌、蜡样芽孢杆菌或其他C. parvum，P23.表达为可溶性scFv-gIIIp构建体的克隆能够检测C. parvum天然蛋白质和子孢子。从幼稚文库中淘选是分离和鉴定具有用于病原体检测和免疫治疗的潜力的重组抗体的有用方法。(C)2010年爱思唯尔公司All rights reserved.
Two human single chain variable fragment (scFv) libraries were used to select clones that bound to the surface glycoprotein S16 of Cryptosporidium parvum. Panning of the Tomlinson libraries I and J resulted in the isolation of nine distinct clones. Of the four clones which had full-length scFv, three contained stop codons. The remaining five clones were truncated, with four missing the heavy chain, and one missing most of the light chain. The full-length clones exhibited better binding to native C. parvum proteins and recombinant S16 than the truncated clones, with the exception of one truncated clone. None of the selected clones cross-reacted with Giardia lamblia, Escherichia coli, Streptococcus pyogenes, Listeria monocytogenes, Bacillus cereus or another immunogenic target of C. parvum, P23. Clones expressed as the soluble scFv-gIIIp construct were able to detect C. parvum native proteins and sporozoites. Panning from naive libraries was an useful method for isolation and identification of recombinant antibodies that have the potential for use in pathogen detection and immunotherapy. (C) 2010 Elsevier Inc. All rights reserved.