Hoe L-N, Wan K-L, Nathan S
Centre for Gene Analysis and Technology, School of Biosciences and Biotechnology, Faculty of Science and Technology, Universiti Kebangsaan Malaysia, 43600 UKM Bangi, Selangor D. E., Malaysia.
Parasitology. 2005 Dec;131(Pt 6):759-68. doi: 10.1017/S0031182005008450.
The protozoan parasite Toxoplasma gondii produces a family of microneme proteins that are thought to play diverse roles in aiding the parasite's intracellular existence. Among these, TgMIC2 has a putative function in parasite adhesion to the host cell to initiate the invasion process. The invasion process may be localized and inhibited by monoclonal antibodies against the protein(s) involved. Here we report on the construction of a phage-displayed single-chain variable fragment (scFv) library from mice immunized with whole T. gondii parasites. The library was subsequently panned against recombinant TgMIC2 (rpTgMIC2) and 2 different groups of antibody clones were obtained, based on fingerprinting and sequencing data. The expressed recombinant scFv antibody was able to recognize rpTgMIC2 in a Western blot detection experiment. These results show that the phage display technology allows quick and effective production of monoclonal antibodies against parasite antigens. By panning the scFv-displayed library, we should be able to obtain a plethora of multi-functional scFv antibodies towards T. gondii proteins.
原生动物寄生虫刚地弓形虫产生一族微线体蛋白，这些蛋白被认为在帮助寄生虫的细胞内存活中发挥多种作用。其中，TgMIC2在寄生虫黏附宿主细胞以启动入侵过程中具有假定功能。入侵过程可能会被针对相关蛋白的单克隆抗体局部化并抑制。在此，我们报道了用完整的刚地弓形虫寄生虫免疫小鼠构建噬菌体展示单链可变片段（scFv）文库的过程。随后，基于指纹图谱和测序数据，将该文库针对重组TgMIC2（rpTgMIC2）进行淘选，获得了2组不同的抗体克隆。在蛋白质印迹检测实验中，表达的重组scFv抗体能够识别rpTgMIC2。这些结果表明，噬菌体展示技术能够快速有效地产生针对寄生虫抗原的单克隆抗体。通过淘选展示scFv的文库，我们应该能够获得大量针对刚地弓形虫蛋白的多功能scFv抗体。
