Institute for Research in Molecular Medicine (INFORMM), Universiti Sains Malaysia, Minden 11800, Penang, Malaysia.
Analytical Biochemistry Research Centre, Universiti Sains Malaysia, Minden 11800, Penang, Malaysia.
Int J Mol Sci. 2017 Nov 22;18(11):2376. doi: 10.3390/ijms18112376.
Helminth parasite infections are significantly impacting global health, with more than two billion infections worldwide with a high morbidity rate. The complex life cycle of the nematodes has made host immune response studies against these parasites extremely difficult. In this study, we utilized two phage antibody libraries; the immune and naïve library were used to identify single chain fragment variable (scFv) clones against a specific filarial antigen (BR1). The V-gene analysis of isolated scFv clones will help shed light on preferential VDJ gene segment usage against the filarial BR1 antigen in healthy and infected states. The immune library showed the usage of both lambda and kappa light chains. However, the naïve library showed preferential use of the lambda family with different amino acid distributions. The binding characteristics of the scFv clones identified from this work were analyzed by immunoassay and immunoaffinity pull down of BR1. The work highlights the antibody gene usage pattern of a naïve and immune antibody library against the same antigen as well as the robust nature of the enriched antibodies for downstream applications.
寄生虫感染对全球健康有重大影响，全世界有超过 20 亿人感染，发病率很高。线虫的复杂生命周期使得针对这些寄生虫的宿主免疫反应研究变得极其困难。在这项研究中，我们利用了两个噬菌体抗体文库；免疫文库和原始文库被用来针对特定的丝虫抗原（BR1）鉴定单链片段可变（scFv）克隆。分离的 scFv 克隆的 V 基因分析将有助于阐明针对健康和感染状态下丝虫 BR1 抗原的优先 VDJ 基因片段使用情况。免疫文库显示了 lambda 和 kappa 轻链的使用。然而，原始文库显示了对 lambda 家族的优先使用，具有不同的氨基酸分布。通过免疫测定和 BR1 的免疫亲和下拉分析，对从这项工作中鉴定出的 scFv 克隆的结合特性进行了分析。这项工作突出了针对同一抗原的原始和免疫抗体文库的抗体基因使用模式，以及富集抗体在下游应用中的强大性质。
