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Thromb Haemost 2000; 83(06): 874-881
DOI: 10.1055/s-0037-1613936
Commentary
Schattauer GmbH

A Monoclonal Antibody Raised against Human β-factor XIIa which also Recognizes α-factor XIIa but not Factor XII or Complexes of Factor XIIa with C1 Esterase Inhibitor

M. Peter Esnouf
1   From the Nuffield Department of Clinical Biochemistry, University of Oxford, The Radcliffe Infirmary, Oxford, UK
,
Annette I. Burgess
1   From the Nuffield Department of Clinical Biochemistry, University of Oxford, The Radcliffe Infirmary, Oxford, UK
,
Alister W. Dodds
2   MRC Immunochemistry Unit, Oxford, UK
,
Anna F. Sarphie
1   From the Nuffield Department of Clinical Biochemistry, University of Oxford, The Radcliffe Infirmary, Oxford, UK
,
George J. Miller
3   MRC Epidemiology and Medical Care Unit, The Wolfson Institute of Preventive Medicine, London, UK
› Author Affiliations
Further Information

Publication History

Received 19 July 1999

Accepted after resubmission 29 January 2000

Publication Date:
14 December 2017 (online)

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Summary

A monoclonal antibody (mAb 2/215) against human β-factor XIIa (β-FXIIa), was shown by equilibrium binding studies to have a high affinity for α-factor XIIa (α-FXIIa) (Kd 1.8 nM) and β-FXIIa (Kd 0.65 nM) but no detectable reaction with FXII zymogen or α - esterase inhibitor (C1-INH) complex. Surface plasmon resonance studies showed that the mAb 2/215 bound to immobilized α-FXIIa with high affinity (KD 3.93 ± 1.46 × 10−11 M). Western blots employing mAb 2/215 indicated that human plasma contained small amounts of α-FXIIa but no β-FXIIa. mAb 2/215 did not inhibit the amidolytic activity of β-FXIIa and protected β-FXIIa from inhibition by C1-INH. The recovery by ELISA ,employing mAb 2/215 as the capture antibody, of α-FXIIa added to plasma was 11.3%, 42% after inhibition of α-FXIIa with 3:4dichloroisocoumarin, and 82% when 0.5% TritonX100 was added to the assay. Gel filtration showed that the majority of plasma α-FXIIa existed as a complex (Mr ∼170000). This distinctive mAb increases the capacity to study the contact system in health and disease.

Key words

Monoclonal antibody - factor XIIa - ELISA - affinity binding studies
 

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