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首页> 外文期刊>Clinical and diagnostic laboratory immunology >Selection of Recombinant, Library-Derived Antibody Fragments against p24 for Human Immunodeficiency Virus Type 1 Diagnostics
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Selection of Recombinant, Library-Derived Antibody Fragments against p24 for Human Immunodeficiency Virus Type 1 Diagnostics

机译：针对人类免疫缺陷病毒1型诊断的p24重组，库衍生抗体片段的选择

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摘要

By application of combinatorial library technology, we generated the first recombinant antibody fragments directed against the major capsid protein p24 of human immunodeficiency virus type 1 (HIV-1). A library of single-chain Fv fragments (scFvs) was constructed by using the antibody variable-region (V) genes of B cells derived from the spleen of a viral lysate-immunized mouse. Antibodies were selected by panning or by enrichment with biotinylated antigen, yielding four different families of antibody fragments. The different types of scFvs were characterized by affinity measurements, by antigen recognition on Western blots, and by pepscan analysis. The epitope of one of the scFvs is located near the residues involved in CypA binding, thereby making it an attractive candidate for therapeutic applications. Comparison of the V gene sequence of this scFV with that of a previously described monoclonal antibody reactive against this immunodominant epitope revealed the usage of the identical combination of VH and Vκ regions. Thus, this is one of the rare examples in which the original combination in a library-derived antibody fragment was retrieved. After appropriate affinity and format improvements, the best of our recombinant scFvs may form the basis for a sensitive p24 assay as a measure of viral load. In addition, anti-p24 scFvs could be expressed as intracellular antibodies (intrabodies) to aid in the treatment of HIV infections.
机译：通过应用组合文库技术，我们产生了第一个针对人类免疫缺陷病毒1型（HIV-1）主要衣壳蛋白p24的重组抗体片段。通过使用源自病毒裂解物免疫小鼠脾脏的B细胞的抗体可变区（V）基因，构建了单链Fv片段（scFvs）库。通过淘选或通过富集生物素化抗原来选择抗体，从而产生四个不同的抗体片段家族。通过亲和力测量，Western印迹上的抗原识别和胃蛋白酶分析来表征不同类型的scFv。 scFv之一的表位位于与CypA结合所涉及的残基附近，从而使其成为用于治疗应用的有吸引力的候选者。该scFV的V基因序列与先前描述的对该免疫优势表位有反应性的单克隆抗体的V基因序列的比较表明，使用了V H 和V κ区域的相同组合。因此，这是罕见的例子之一，其中检索了文库衍生的抗体片段中的原始组合。经过适当的亲和力和形式改进后，我们最好的重组scFv可能构成敏感的p24测定法的基础，以此作为病毒载量的量度。此外，抗p24 scFvs可以表达为细胞内抗体（体内抗体），以帮助治疗HIV感染。

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