NDLI: Isolation of anti-CD22 Fv with high affinity by Fv display on human cells

Content Provider	PubMed Central
Author	Ho, Mitchell Nagata, Satoshi Pastan, Ira
Copyright Year	2006
Abstract	In vitro antibody affinity maturation has generally been achieved by display of mouse or human antibodies on the surface of microorganisms (phage, bacteria, and yeast). However, problems with protein folding, posttranslational modification, and codon usage still limit the number of improved antibodies that can be obtained. An ideal system would select and improve antibodies in a mammalian cell environment where they are naturally made. Here we show that human embryonic kidney 293T cells that are widely used for transient protein expression can be used for cell surface display of single-chain Fv antibodies for affinity maturation. In a proof-of-concept experiment, cells expressing a rare mutant antibody with higher affinity were enriched 240-fold by a single-pass cell sorting from a large excess of cells expressing WT antibody with a slightly lower affinity. Furthermore, we successfully obtained a highly enriched mutant with increased binding affinity for CD22 after a single selection of a combinatory library randomizing an intrinsic antibody hotspot. Important features are that one display selection cycle requires only 1 week, and transfection of cells in a single 100-mm dish produces 107 individual clones so that a repertoire of 109 is feasible under current experimental conditions.
Related Links	http://dx.doi.org/10.1073/pnas.0603653103
Ending Page	9642
Page Count	6
Starting Page	9637
File Format	PDF
ISSN	00278424
e-ISSN	10916490
Journal	Proceedings of the National Academy of Sciences of the United States of America
Issue Number	25
Volume Number	103
Language	English
Publisher	National Academy of Sciences
Publisher Date	2006-06-20
Access Restriction	Open
Rights Holder	National Academy of Sciences
Subject Keyword	General Research in Higher Education
Content Type	Text
Resource Type	Article
Subject	Multidisciplinary

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Correction for Ho et al., Isolation of anti-CD22 Fv with high affinity by Fv display on human cells

Isolation of anti-CD22 Fv with high affinity by Fv display on human cells.

Isolation of a high-affinity stable single-chain Fv specific for mesothelin from DNA-immunized mice by phage display and construction of a recombinant immunotoxin with anti-tumor activity

Specificity grafting of human antibody frameworks selected from a phage display library: generation of a highly stable humanized anti‐CD22 single‐chain Fv fragment

Isolation of human monoclonal antibodies by mammalian cell display

An extracatalytic function of CD45 in B cells is mediated by CD22

Isolation of a high-affinity stable single-chain Fv specific for mesothelin from DNA-immunized mice by phage display and construction of a recombinant immunotoxin with anti-tumor activity.

The use of mRNA display to select high-affinity protein-binding peptides

Quantitative analysis of the effect of the mutation frequency on the affinity maturation of single chain Fv antibodies

Isolation of anti-CD22 Fv with high affinity by Fv display on human cells

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Correction for Ho et al., Isolation of anti-CD22 Fv with high affinity by Fv display on human cells

Isolation of anti-CD22 Fv with high affinity by Fv display on human cells.

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