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Efficient construction of a highly useful phage-displayed human antibody repertoire.

Abstract：

:We have constructed a highly useful phage-displayed human antibody repertoire with limited cloning efforts. Our strategy was to maximize diversity during the first steps of library construction through the use of various lymphoid sources from several donors, inclusion of different immunoglobulin isotypes, and performance of multiple separate amplification reactions with all possible combinations within a complex primer set. The resulting variable region collections were cloned to form a moderate size library, composed by 4.25x10(8) single chain antibody fragments. This repertoire was successfully used to retrieve binders to seven model antigens: six proteins and one 12 aa peptide. Binding affinities reached nanomolar and even subnanomolar range. Sequence diversity and V-gene usage variability among binders were proven. Our approach was not focused on absolute library size, but on a high quality sampling of variable regions from the human antibody repertoire.

journal_name

Biochem Biophys Res Commun

journal_title

Biochemical and biophysical research communications

authors

Rojas G,Lamdan H,Padron S,Munoz Y,Ayala M,Gavilondo JV

doi

10.1016/j.bbrc.2005.09.005

subject

Has Abstract

pub_date

2005-11-04 00:00:00

pages

1207-13

issue

4

eissn

0006-291X

issn

1090-2104

pii

S0006-291X(05)01997-2

journal_volume

336

pub_type

杂志文章

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