Panning Of Human Antibody Against Tetanus Toxin by Phage Display Method
Subject Areas :Hamideh Rouhani Nejad 1 , Jalil Fallah Mehrabadi 2 , Masomeh Bazaz 3
1 - پژوهشکده علوم و فناوری زیستی، دانشگاه صنعتی مالک اشتر، تهران. ایران.
2 - گروه زیست شناسی، دانشکده علوم پایه، دانشگاه قم، قم. ایران
3 - پژوهشکده علوم و فناوری زیستی، دانشگاه صنعتی مالک اشتر، تهران. ایران
Keywords: Biopanning, Tetanus, Phage Display,
Abstract :
Inroduction and Objective: Tetanus toxin is one of the most toxic substrate acting on the central nerve system causing severe muscle contraction. Rapid diagnosis and treatment of tetanus neurotoxins are crucial. Antibodies are important tools in research, diagnosis and treatment of diseases. Fully human mAbs because of minimum immunogenic reaction and high performance are important for treatment.Antibodies affinity maturation has facilitated their application in medicine and research projects.Methods: The principle of this study was to screening in immune antibody library for isolation high specificity antibody against tetanus toxin. Library was to construct with pSEX81 expression vector, and then pSEX81 was extracted from library.For screening in this library, biopanning was performed by phage display method.Results: Helper phage M13K07 was prepared at the titer of 1013pfu/ml. The tetanus toxin was coated in microtiter plate and biopanning process was done using phage particles. The exactness of enrichment process was confirmed by phage ELISA. Single clones wereisolated from biopanning cycle with highest optical density. To isolate the tetanus specificantibodies, first vector extraction was done from library. After that, five rounds of biopanning were done. The strongest enrichment occurred at the third cycle of panning and confirmed the existence of high affinity tetanus antibodies. A total 50 single colonies randomly selected from third cycle of biopanning and 20 clones with strong binding were selected and sequenced.Conclusion: In this study, our aim was the screening of tetanus antibodies by phage display selection method. The antibody fragments isolated from immune libraries applications finds in the diagnostic and treatment tools.
