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Efficient recovery of high-affinity antibodies from a single-chain Fab yeast display library.

Abstract：

:Yeast display is a powerful technology for the isolation of monoclonal antibodies (mAbs) against a target antigen. Antibody libraries have been displayed on the surface of yeast as both single-chain variable fragment (scFv) and antigen binding fragment (Fab). Here, we combine these two formats to display well-characterized mAbs as single-chain Fabs (scFabs) on the surface of yeast and construct the first scFab yeast display antibody library. When expressed on the surface of yeast, two out of three anti-human immunodeficiency virus (HIV)-1 mAbs bound with higher affinity as scFabs than scFvs. Also, the soluble scFab preparations exhibited binding and neutralization profiles comparable to that of the corresponding Fab fragments. Display of an immune HIV-1 scFab library on the surface of yeast, followed by rounds of sorting against HIV-1 gp120, allowed for the selection of 13 antigen-specific clones. When the same cDNA was used to construct the library in an scFv format, a similar number but a lower affinity set of clones were selected. Based on these results, yeast-displayed scFab libraries can be constructed and selected with high efficiency, characterized without the need for a reformatting step, and used to isolate higher-affinity antibodies than scFv libraries.

journal_name

J Mol Biol

journal_title

Journal of molecular biology

authors

Walker LM,Bowley DR,Burton DR

doi

10.1016/j.jmb.2009.04.019

subject

Has Abstract

pub_date

2009-06-05 00:00:00

pages

365-75

issue

2

eissn

0022-2836

issn

1089-8638

pii

S0022-2836(09)00449-5

journal_volume

389

pub_type

杂志文章

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