Norbury Luke J, Basałaj Katarzyna, Bąska Piotr, Kalinowska Alicja, Zawistowska-Deniziak Anna, Yap Huan Yong, Wilkowski Przemysław, Wesołowska Agnieszka, Wędrychowicz Halina
Witold Stefański Institute of Parasitology, Polish Academy of Sciences, Twarda 51/55, 00-818, Warsaw, Poland; School of Science, Royal Melbourne Institute of Technology (RMIT) University, Bundoora, 3083, Victoria, Australia.
Witold Stefański Institute of Parasitology, Polish Academy of Sciences, Twarda 51/55, 00-818, Warsaw, Poland.
Exp Parasitol. 2019 Mar;198:87-94. doi: 10.1016/j.exppara.2019.02.001. Epub 2019 Feb 11.
Phage display technology to produce recombinant monoclonal antibodies or antibody fragments permits the identification of sought after antibodies in short time frames at low cost along with direct and rapid selection for antibody characteristics. Monoclonal antibodies can facilitate the identification and characterisation of parasite molecules that function at the host-parasite interface to help understand at the molecular level the biology of the parasite and disease progression, which often leads to new drug targets, diagnostic antigens or vaccine candidates. The trematode Fasciola hepatica is an important veterinary and human parasite. In this work, we infected rats with F. hepatica and amplified the generated antibody repertoire to produce a single-chain variable fragment (scFv) phage display library. The library was used to identify a scFv that recognises cathepsin L1, a major component of the adult parasites excretory/secretory material and an important vaccine candidate. This is the first report of the construction of a phage display antibody library from a F. hepatica infected host, and also the first instance such a library has been used to identify an affinity-matured monoclonal antibody fragment that binds to a F. hepatica antigen. The scFv library and methods detailed should facilitate future research characterising F. hepatica antigens.
利用噬菌体展示技术生产重组单克隆抗体或抗体片段,能够在短时间内以低成本鉴定出所需抗体,同时可直接快速筛选抗体特性。单克隆抗体有助于鉴定和表征在宿主 - 寄生虫界面发挥作用的寄生虫分子,从而在分子水平上帮助理解寄生虫生物学和疾病进展情况,这往往会带来新的药物靶点、诊断抗原或疫苗候选物。肝片吸虫是一种重要的兽医和人类寄生虫。在这项研究中,我们用肝片吸虫感染大鼠,并扩增产生的抗体库以构建单链可变片段(scFv)噬菌体展示文库。该文库用于鉴定一种识别组织蛋白酶L1的scFv,组织蛋白酶L1是成年寄生虫排泄/分泌物质的主要成分,也是一种重要的疫苗候选物。这是首次报道从感染肝片吸虫的宿主构建噬菌体展示抗体文库,也是首次利用这样的文库鉴定出与肝片吸虫抗原结合的亲和力成熟的单克隆抗体片段。所详述的scFv文库和方法应有助于未来对肝片吸虫抗原的研究。