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Thromb Haemost 2001; 85(03): 379-389
DOI: 10.1055/s-0037-1615593
Rapid Communications
Schattauer GmbH

Generation of a Humanized, High Affinity Anti-tissue Factor Antibody for Use as a Novel Antithrombotic Therapeutic

Leonard Presta
,
Paul Sims
2   BioAnalytical Technology
,
Y. Gloria Meng
2   BioAnalytical Technology
,
Paul Moran
3   Cardiovascular Research
,
Sherron Bullens
3   Cardiovascular Research
,
Stuart Bunting
3   Cardiovascular Research
,
Jill Schoenfeld
3   Cardiovascular Research
,
David Lowe
3   Cardiovascular Research
,
Jadine Lai
,
Patricia Rancatore
4   Recovery Sciences
,
Mark Iverson
4   Recovery Sciences
,
Amy Lim
4   Recovery Sciences
,
Vanessa Chisholm
5   Molecular Biology
,
Robert F. Kelley
6   Protein Engineering, Genentech Inc., South San Francisco, CA, USA
,
Markus Riederer
7   Preclinical Research Department, F. Hoffmann-La Roche Ltd., Basel, Switzerland
,
Daniel Kirchhofer
3   Cardiovascular Research
› Author Affiliations
Further Information

Publication History

Received 21 June 2000

Accepted after revision 14 September 2000

Publication Date:
08 December 2017 (online)

Also available at
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Summary

Blocking the cofactor function of human tissue factor may be beneficial in various coagulation-mediated diseases. The murine antibody D3 binds to the membrane proximal substrate interaction region of human tissue factor and blocks tissue factor function even in the presence of bound factor VIIa. The cloned murine D3 antibody was humanized and affinity matured by exchanging amino acids in the complementarity determining regions as well as in the antibody framework. The humanized antibody, D3H44, bound to tissue factor with a 100-fold increased affinity (KD 0.1 nM) as compared to the original murine and chimeric versions. Depending on the particular disease, different pharmacokinetic properties of the antibody may be required and, therefore, several antibody variants – F(ab), F(ab’)2, IgG2, IgG4 and IgG4b – were generated. In vitro, the humanized D3 antibodies displayed potent inhibition of plasma clotting and tissue factor: factor VIIa-mediated activation of factors IX and X (e. g. D3H44-F(ab’)2, IC50 (F.X) 47 pM). In addition, D3H44-F(ab’)2 completely prevented fibrin deposition in a human ex vivo thrombosis model under venous blood flow conditions (IC50 37 nM). The humanized D3 antibodies may be utilized for treatment of cardiovascular diseases which involve tissue factor activity, e. g. acute coronary syndrome and venous thrombosis.

Keywords

Tissue factor - coagulation factor VII - humanization - anticoagulants - thrombosis
 

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