Omar Noorsharmimi, Hamidon Nurul Hamizah, Yunus Muhammad Hafiznur, Noordin Rahmah, Choong Yee Siew, Lim Theam Soon
Institute for Research in Molecular Medicine, Universiti Sains Malaysia, Minden, Penang, Malaysia.
Analytical Biochemistry Research Centre, Universiti Sains Malaysia, Penang, Malaysia.
Biotechnol Appl Biochem. 2018 May;65(3):346-354. doi: 10.1002/bab.1591. Epub 2017 Sep 14.
Phage display has been applied successfully as a tool for the generation of monoclonal antibodies (mAbs). Naive antibody libraries are unique as they are able to overcome several limitations associated with conventional mAb generation methods like the hybridoma technology. Here, we performed an in vitro selection and generation of Fab antibodies against Brugia malayi SXP protein (BmSXP), a recombinant antigen for the detection of lymphatic filariasis. We developed a naïve multi ethnic Fab antibody library with an estimated diversity of 2.99 × 10 . The antibody library was used to screen for mAbs against BmSXP recombinant antigen. Soluble monoclonal Fab antibodies against BmSXP were successfully isolated from the naïve library. The Fab antibodies obtained were expressed and analyzed to show its binding capability. The diversity obtained from a pool of donors from various ethnic groups allowed for a diverse antibody library to be generated. The mAbs obtained were also functional in soluble form, which makes it useful for further downstream applications. We believe that the Fab mAbs are valuable for further studies and could also contribute to improvements in the diagnosis of filariasis.
噬菌体展示已成功应用于单克隆抗体（mAb）的产生。天然抗体文库具有独特性，因为它们能够克服与传统单克隆抗体制备方法（如杂交瘤技术）相关的一些局限性。在此，我们针对马来布鲁线虫SXP蛋白（BmSXP）进行了Fab抗体的体外筛选和产生，BmSXP是一种用于检测淋巴丝虫病的重组抗原。我们构建了一个估计多样性为2.99×10的天然多民族Fab抗体文库。该抗体文库用于筛选针对BmSXP重组抗原的单克隆抗体。成功从天然文库中分离出针对BmSXP的可溶性单克隆Fab抗体。对获得的Fab抗体进行表达和分析以显示其结合能力。来自不同种族供体库的多样性使得能够构建一个多样化的抗体文库。获得的单克隆抗体以可溶性形式也具有功能，这使其可用于进一步的下游应用。我们认为，Fab单克隆抗体对进一步研究具有重要价值，也有助于改进丝虫病的诊断。
