CHO expression of a novel human recombinant IgG1 anti-RhD antibody isolated by phage display.

Details

Serval ID
serval:BIB_16540
Type
Article: article from journal or magazin.
Collection
Publications
Institution
Title
CHO expression of a novel human recombinant IgG1 anti-RhD antibody isolated by phage display.
Journal
British Journal of Haematology
Author(s)
Miescher S., Zahn-Zabal M., De Jesus M., Moudry R., Fisch I., Vogel M., Kobr M., Imboden M.A., Kragten E., Bichler J., Mermod N., Stadler B.M., Amstutz H., Wurm F.
ISSN
0007-1048 (Print)
ISSN-L
0007-1048
Publication state
Published
Issued date
2000
Volume
111
Number
1
Pages
157-166
Language
english
Notes
Publication types: Journal Article ; Research Support, Non-U.S. Gov't
Publication Status: ppublish
Abstract
Replacement of the hyperimmune anti-Rhesus (Rh) D immunoglobulin, currently used to prevent haemolytic disease of the newborn, by fully recombinant human anti-RhD antibodies would solve the current logistic problems associated with supply and demand. The combination of phage display repertoire cloning with precise selection procedures enables isolation of specific genes that can then be inserted into mammalian expression systems allowing production of large quantities of recombinant human proteins. With the aim of selecting high-affinity anti-RhD antibodies, two human Fab libraries were constructed from a hyperimmune donor. Use of a new phage panning procedure involving bromelin-treated red blood cells enabled the isolation of two high-affinity Fab-expressing phage clones. LD-6-3 and LD-6-33, specific for RhD. These showed a novel reaction pattern by recognizing the D variants D(III), D(IVa), D(IVb), D(Va), D(VI) types I and II. D(VII), Rh33 and DFR. Full-length immunoglobulin molecules were constructed by cloning the variable regions into expression vectors containing genomic DNA encoding the immunoglobulin constant regions. We describe the first, stable, suspension growth-adapted Chinese hamster ovary (CHO) cell line producing a high affinity recombinant human IgG1 anti-RhD antibody adapted to pilot-scale production. Evaluation of the Fc region of this recombinant antibody by either chemiluminescence or antibody-dependent cell cytotoxicity (ADCC) assays demonstrated macrophage activation and lysis of red blood cells by human lymphocytes. A consistent source of recombinant human anti-RhD immunoglobulin produced by CHO cells is expected to meet the stringent safety and regulatory requirements for prophylactic application.
Keywords
Animals, Bacteriophages, Base Sequence, Biotechnology/methods, Bromelains/pharmacology, CHO Cells, Cloning, Molecular, Cricetinae, Erythrocytes, Humans, Immunoglobulin Fab Fragments/genetics, Immunoglobulin Fab Fragments/isolation & purification, Immunoglobulin G/genetics, Molecular Sequence Data, Recombinant Proteins/metabolism, Rh Isoimmunization/prevention & control, Rho(D) Immune Globulin/metabolism
Pubmed
Web of science
Open Access
Yes
Create date
19/11/2007 13:09
Last modification date
20/08/2019 13:45
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