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On the influence of vector design on antibody phage display

Paper ID Volume ID Publish Year Pages File Format Full-Text
25512 43578 2007 12 PDF Available
Title
On the influence of vector design on antibody phage display
Abstract

Phage display technology is an established technology particularly useful for the generation of monoclonal antibodies (mAbs). The isolation of phagemid-encoded mAb fragments depends on several features of a phage preparation. The aims of this study were to optimize phage display vectors, and to ascertain if different virion features can be optimized independently of each other.Comparisons were made between phagemid virions assembled by g3p-deficient helper phage, Hyperphage, Ex-phage or Phaberge, or corresponding g3p-sufficient helper phage, M13K07. All g3p-deficient helper phage provided a similar level of antibody display, significantly higher than that of M13K07. Hyperphage packaged virions at least 100-fold more efficiently than did Ex-phage or Phaberge. Phaberge's packaging efficiency improved by using a SupE strain.Different phagemids were also compared. Removal of a 56 base pair fragment from the promoter region resulted in increased display level and increased virion production. This critical fragment encodes a lacZ′-like peptide and is also present in other commonly used phagemids.Increasing display level did not show statistical correlation with phage production, phage infectivity or bacterial growth rate. However, phage production was positively correlated to phage infectivity.In summary, this study demonstrates simultaneously optimization of multiple and independent features of importance for phage selection.

Keywords
A600, absorbance at 600 nm; CFU, colony forming units; ELISA, enzyme-linked immunosorbent assay; Fab, antigen-binding fragment; g3p, gene III protein; lacZ, beta galactosidase gene; mAb, monoclonal antibody; SupE, mutant phenotype conferring read-through
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On the influence of vector design on antibody phage display
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Publisher
Database: Elsevier - ScienceDirect
Journal: Journal of Biotechnology - Volume 127, Issue 4, 20 January 2007, Pages 626–637
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us