A two-phagemid system for the creation of non-phage displayed antibody libraries approaching one trillion members

Marc Ostermeier; Stephen J. Benkovic

doi:10.1016/S0022-1759(99)00245-8

A two-phagemid system for the creation of non-phage displayed antibody libraries approaching one trillion members

Marc Ostermeier, Stephen J. Benkovic

Chemistry

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9 Scopus citations

Abstract

We have designed a two-phagemid system for the construction of very large non-phage displayed Fab antibody libraries in E. coli approaching 10¹² members. The system can accommodate both periplasmic and cytoplasmic Fab expression and should prove useful for the direct selection of functional antibodies by genetic techniques. We successfully alleviate problems of Fab vector instability and report a set of improved 5' primers for the amplification of mouse Ig V(H) repertoires from mouse spleen. These primers have no more than one mismatch in the last 11 bases for >95% of mouse Ig V(H) genes and minimize the amount of N-terminal amino acid changes while maintaining the flexibility of periplasmic or cytoplasmic antibody expression in E. coli. Copyright (C) 2000 Elsevier Science B.V.

Original language	English (US)
Pages (from-to)	175-186
Number of pages	12
Journal	Journal of Immunological Methods
Volume	237
Issue number	1-2
DOIs	https://doi.org/10.1016/S0022-1759(99)00245-8
State	Published - Apr 3 2000

All Science Journal Classification (ASJC) codes

Immunology and Allergy
Immunology

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10.1016/S0022-1759(99)00245-8

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title = "A two-phagemid system for the creation of non-phage displayed antibody libraries approaching one trillion members",

abstract = "We have designed a two-phagemid system for the construction of very large non-phage displayed Fab antibody libraries in E. coli approaching 1012 members. The system can accommodate both periplasmic and cytoplasmic Fab expression and should prove useful for the direct selection of functional antibodies by genetic techniques. We successfully alleviate problems of Fab vector instability and report a set of improved 5' primers for the amplification of mouse Ig V(H) repertoires from mouse spleen. These primers have no more than one mismatch in the last 11 bases for >95% of mouse Ig V(H) genes and minimize the amount of N-terminal amino acid changes while maintaining the flexibility of periplasmic or cytoplasmic antibody expression in E. coli. Copyright (C) 2000 Elsevier Science B.V.",

author = "Marc Ostermeier and Benkovic, {Stephen J.}",

note = "Funding Information: We would like to thank Dr D.R. Burton for providing plasmid pTC01 and Dr K.F Jensen for providing strains S{\O}4038 and S{\O}6355. This work was supported by a NIH postdoctoral fellowship (1 F32 GM18560-01) to M.O.",

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doi = "10.1016/S0022-1759(99)00245-8",

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T1 - A two-phagemid system for the creation of non-phage displayed antibody libraries approaching one trillion members

AU - Ostermeier, Marc

AU - Benkovic, Stephen J.

N1 - Funding Information: We would like to thank Dr D.R. Burton for providing plasmid pTC01 and Dr K.F Jensen for providing strains SØ4038 and SØ6355. This work was supported by a NIH postdoctoral fellowship (1 F32 GM18560-01) to M.O.

PY - 2000/4/3

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N2 - We have designed a two-phagemid system for the construction of very large non-phage displayed Fab antibody libraries in E. coli approaching 1012 members. The system can accommodate both periplasmic and cytoplasmic Fab expression and should prove useful for the direct selection of functional antibodies by genetic techniques. We successfully alleviate problems of Fab vector instability and report a set of improved 5' primers for the amplification of mouse Ig V(H) repertoires from mouse spleen. These primers have no more than one mismatch in the last 11 bases for >95% of mouse Ig V(H) genes and minimize the amount of N-terminal amino acid changes while maintaining the flexibility of periplasmic or cytoplasmic antibody expression in E. coli. Copyright (C) 2000 Elsevier Science B.V.

AB - We have designed a two-phagemid system for the construction of very large non-phage displayed Fab antibody libraries in E. coli approaching 1012 members. The system can accommodate both periplasmic and cytoplasmic Fab expression and should prove useful for the direct selection of functional antibodies by genetic techniques. We successfully alleviate problems of Fab vector instability and report a set of improved 5' primers for the amplification of mouse Ig V(H) repertoires from mouse spleen. These primers have no more than one mismatch in the last 11 bases for >95% of mouse Ig V(H) genes and minimize the amount of N-terminal amino acid changes while maintaining the flexibility of periplasmic or cytoplasmic antibody expression in E. coli. Copyright (C) 2000 Elsevier Science B.V.

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A two-phagemid system for the creation of non-phage displayed antibody libraries approaching one trillion members

Abstract

All Science Journal Classification (ASJC) codes

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