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

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

Miescher, S.
•
Zahn-Zabal, M.
•
De Jesus, M.  
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2000
British journal of haematology

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.

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Type
research article
DOI
10.1046/j.1365-2141.2000.02322.x
Web of Science ID

WOS:000165511700021

PubMed ID

11091196

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.
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Date Issued

2000

Published in
British journal of haematology
Volume

111

Issue

1

Start page

157

End page

66

Subjects

Animals

•

Bacteriophages

•

Base Sequence

•

Biotechnology/*methods

•

Bromelains/pharmacology

•

CHO Cells

•

Cloning

•

Molecular

•

Cricetinae

•

Erythrocytes

•

Humans

•

Immunoglobulin Fab Fragments/*genetics/isolation & purification

•

Immunoglobulin G/*genetics

•

Molecular Sequence Data

•

Recombinant Proteins/metabolism

•

Rh Isoimmunization/prevention & control

•

Rho(D) Immune Globulin/*metabolism

Note

ZLB Central Laboratory, Swiss Red Cross, Inselspital, Bern, Switzerland. sylvia.miescher@insel.ch

Journal Article

Research Support, Non-U.S. Gov't

England

Editorial or Peer reviewed

REVIEWED

Written at

EPFL

EPFL units
LBTC  
Available on Infoscience
June 5, 2007
Use this identifier to reference this record
https://infoscience.epfl.ch/handle/20.500.14299/7585
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