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| Screening and Indetification of Anti-rabies Virus Single Chain Fragments Antiboy from Human Phage Display Library |
| CHEN Ji-jun1, MAO Xiao-yan1, QIAO Yu-ling1, BI Si-ying2 |
1 Lanzhou Institute of Biological Products Co,. Ltd;
2 Shandong Bi Bo Biological Technology Co,.Ltd |
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Abstract Objective: Construct a human anti-rabies virus single-chain fragments (scFV) antibody phage antibody library. Screening, expression and indeticification of human single chain(scFv) with in vivo anti-rabies neutralizing activity by phage display. Methods: Based on phage display libraries which originate from the peripheral blood of 21 high titers of healthy donors vaccinated rabies vaccines, purified rbies virus was coated on immune tubes. Three round screens was executed. The postivie rate of monclone phage was detected by ELISA. Sequences of positive clone were analysed by Vbase2 and DNAStar. E.coli BL21(DE3) recombinant expression system were constructed. ScFv was expressed by IPTG induction and purified by Ni affinity chromatography. Determined the vivo neutralizing activity of scFv by RFFIT and detected the coss-binding capacity with 3aG, CVS, CTN and PV rabies virus stain by indrect ELISA. Results: 40 scFv sequences were obtained. 7 scFv-BL21(DE3) recombinant expression system were constructed. 3/7 scFv-BL21(DE3) were soluble expression. 4/7 scFv had in vivo neutralizing activity more than 500IU/mg. All of the scFv had evidented coss-binding capcity with 3aG, PV and CTN starin. Conclusion: ScFv were obtained for anti-rabies virus from phage antibody libirary. RFFIT results show that expressed scFvs have in vivo anti-rabies virus neturilizing activity.
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Received: 28 August 2013
Published: 25 November 2013
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