Zhi-Chao Zhang1, Xue-Jun Hu, Qing Yang. 1. State Key Laboratory of Fine Chemicals, Dalian University of Technology, Dalian 116012, China. zzc1217@sina.com
Abstract
BACKGROUND: A single-chain antibody (ScFv) phage display library was created by cloning antigen-binding regions of V(H) (variable domain) and V(L) gene repertoires as fusion proteins with a minor coat protein of filamentous phage, from which high affinity completely humanized ScFv against PreS1 of hepatitis B virus could be screened and characterized. METHODS: A combinatorial library of phage-display human ScFv genes, which were derived from peripheral blood lymphocytes immunized by peptide PreS1 in vitro, was constructed. The library contained 7 x 10(8) clones. RESULTS: After 3 rounds panning, a high affinity (K=10(-7)-10(-8)mol/L) ScFv specific to PreS1 was obtained. Sequence analysis showed that the V(H) belonged to the V(H)4 family and V(lambda) to V(lambda)4. CONCLUSIONS: The described ScFv may provide a more satisfactory therapy. This application further illustrates that the method of in vitro antigen stimulation is expeditious for the source of human immune antibody library.
BACKGROUND: A single-chain antibody (ScFv) phage display library was created by cloning antigen-binding regions of V(H) (variable domain) and V(L) gene repertoires as fusion proteins with a minor coat protein of filamentous phage, from which high affinity completely humanized ScFv against PreS1 of hepatitis B virus could be screened and characterized. METHODS: A combinatorial library of phage-display humanScFv genes, which were derived from peripheral blood lymphocytes immunized by peptide PreS1 in vitro, was constructed. The library contained 7 x 10(8) clones. RESULTS: After 3 rounds panning, a high affinity (K=10(-7)-10(-8)mol/L) ScFv specific to PreS1 was obtained. Sequence analysis showed that the V(H) belonged to the V(H)4 family and V(lambda) to V(lambda)4. CONCLUSIONS: The described ScFv may provide a more satisfactory therapy. This application further illustrates that the method of in vitro antigen stimulation is expeditious for the source of human immune antibody library.
Authors: Krista M McCutcheon; Julia Gray; Natalie Y Chen; Keyi Liu; Minha Park; Stote Ellsworth; Ralph A Tripp; S Mark Tompkins; Scott K Johnson; Shelly Samet; Lenore Pereira; Lawrence M Kauvar Journal: MAbs Date: 2014-01-08 Impact factor: 5.857