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Thromb Haemost 1988; 60(02): 334-339
DOI: 10.1055/s-0038-1647056
Original Article
Schattauer GmbH Stuttgart

Monoclonal Antibodies Against the Heparin- Dependent Protein C Inhibitor Suitable for Inhibitor Purification and Assay of Inhibitor Complexes

Martin Laurell
1   The Department of Clinical Chemistry, Malmo General Hospital, Malmo, Sweden
,
Timothy H Carlson
1   The Department of Clinical Chemistry, Malmo General Hospital, Malmo, Sweden
,
Johan Stenflo
1   The Department of Clinical Chemistry, Malmo General Hospital, Malmo, Sweden
› Author Affiliations
Further Information

Publication History

Received 18 April 1988

Accepted after revision 22 June 1988

Publication Date:
28 June 2018 (online)

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summary

Two different monoclonal antibodies against the heparin- dependent inhibitor of human activated protein C were produced, using cleaved modified inhibitor for immunization and partially purified inhibitor for screening of the hybridomas, One of the antibodies recognized free and complexed forms of the inhibitor in immunoblotting experiments. The other antibody was used to develop an assay for A PC -PCI inhibitor complexes. Using the assay the formation of complexes was studied in plasma, both in the presence and absence of heparin. The rate of complex formation was similar to that reported previously for the loss of activated protein C amidolytic activity in plasma. The same antibody was also immobilized on Sepharose and used to purify the inhibitor from fresh human plasma. The purified material appeared as two narrowly spaced bands with Mr about 57,000 in SDS-PAGE. The average yield from 1 liter of fresh plasma was 1 mg of inhibitor, The purified inhibitor formed SDS stable complexes with activated protein G and urokinase that could be identified in immunoblots using specific antibodies.

Keywords

Protein C - Protein C inhibitor
 

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