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Expression in Pichia pastoris of a recombinant scFv form of MAb 107, an anti human CD11b integrin antibody

Paper ID Volume ID Publish Year Pages File Format Full-Text
18618 42728 2006 7 PDF Available
Title
Expression in Pichia pastoris of a recombinant scFv form of MAb 107, an anti human CD11b integrin antibody
Abstract

We herein report the production in the methylotrophic yeast Pichia pastoris of a recombinant scFv form of monoclonal antibody 107 [MAb 107]. This antibody is directed to the leukocyte adhesion molecule CR3 (CD11b/CD18) and is specific of the CD11bA domain. MAb 107 is a ligand mimic of CR3 that blocks the adhesion of leukocyte and their transendothelial migration (diapedesis), a feature that confers this MAb a potential anti inflammatory activity. Expression of scFv 107 as a myc-(His)6 secreted fusion protein was achieved in the yeast P. pastoris using plasmid pICZαB. A spontaneous specific cleavage of the myc-His tag was consistently observed regardless of the culture conditions and the addition of different concentrations of casaminoacids in the medium. Loss of the myc-His tag did not affect the binding activity of scFv 107. This loss is due either to a specific proteolysis or to a mechanical cleavage. Optimization of the production of scFv 107 in shake flasks was carried out using an L16 array Taguchi design. A level of purified scFv-myc-His fusion protein estimated at 40 mg/L was reached at pH 6 and a temperature of 27 °C. The production of large amounts of recombinant scFv form of MAb 107 is essential for further molecular studies of the interactions between CR3 and its ligands, mainly those mediated by the CD11b A domain. It will also facilitate the investigation of its anti inflammatory activity in vivo.

Keywords
Pichia pastoris; Taguchi experimental design; Monoclonal antibody; Anti human integrin
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Expression in Pichia pastoris of a recombinant scFv form of MAb 107, an anti human CD11b integrin antibody
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Publisher
Database: Elsevier - ScienceDirect
Journal: Enzyme and Microbial Technology - Volume 38, Issue 5, 2 March 2006, Pages 636–642
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