Generation of recombinant antibody libraries that facilitate highly sensitive monitoring of low molecular weight biomarkers

为了产生针对小分子标志物的高效ScFv突变体，我们构建了一个基于单个主框的半抗原靶向ScFv文库，其中VH和VL亚群是经常用于抗半抗原抗体的两个亚群。然而，该文库未能产生任何实用的结合蛋白，这表明有必要进行反复试验，以选择合适的位置/模式进行突变。因此，我们决定使用链霉亲和素(STIV)作为一种新的支架，并产生了一个STIV突变体文库，每个成员的环结构都有随机突变。经过几轮筛选，我们发现突变克隆获得了与雌二醇或皮质醇结合的能力。

To generate high-performance scFv mutants that are specific to small biomarkers, we constructed a hapten- targeting scFv library based on a single master-frame of which VH and VL subgroups are those frequently used in anti-hapten antibodies. However, this library failed to yield any practical binder, suggesting that repeated trials are necessary to select suitable positions/patterns for mutagenesis. Thus, we decided to use streptavidin (stav) as a new scaffold, and generated a library of stav mutatants, each member of which has random mutations in their loop structures. After several rounds of selection, wefound mutant clones that gained binding abilities to estradiol or cortisol.