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Selection and affinity maturation of human antibodies against rabies virus from a scFv gene library using ribosome display

Paper ID Volume ID Publish Year Pages File Format Full-Text
24405 43513 2009 6 PDF Available
Title
Selection and affinity maturation of human antibodies against rabies virus from a scFv gene library using ribosome display
Abstract

We selected useful antibody fragments against rabies virus from a human single chain variable fragment (scFv) gene library using ribosome display technique. The recombinant rabies virus glycoprotein (RVGp) was used as an antigen to isolate specific scFvs. After five rounds of selection, the analysis demonstrated that scFv–ribosome–mRNA complexes were specifically selected against RVGp. Sequence analysis showed that mutations were introduced at random by PCR among the rounds of selection and variants with high affinity were isolated. The obtained scFvs with high affinity could recognize RVGp specifically and showed binding activity to rabies virus. These scFvs were potential for inclusion in a combination of several human monoclonal antibodies (MAbs) aimed for application in rabies post-exposure prophylaxis. Ribosome display technology is a robust tool for rapid isolation of human antibody fragments, and has exceptional strength in affinity maturation and molecular evolution in vitro.

Keywords
Human antibody; Affinity maturation; Ribosome display; Rabies virus
First Page Preview
Selection and affinity maturation of human antibodies against rabies virus from a scFv gene library using ribosome display
Publisher
Database: Elsevier - ScienceDirect
Journal: Journal of Biotechnology - Volume 144, Issue 4, December 2009, Pages 253–258
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering