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Thromb Haemost 1985; 53(02): 170-175
DOI: 10.1055/s-0038-1661266
Original Article
Schattauer GmbH Stuttgart

Characterization of Two Monoclonal Antibodies Against Human Tissue-Type Plasminogen Activator

Raymond R Schleef
The Department of Immunology, Scripps Clinic and Research Foundation, La Jolla, CA, USA
,
Manjula Sinha
The Department of Immunology, Scripps Clinic and Research Foundation, La Jolla, CA, USA
,
David J Loskutoff
The Department of Immunology, Scripps Clinic and Research Foundation, La Jolla, CA, USA
› Author Affiliations
Further Information

Publication History

Received 21 August 1984

Accepted 29 November 1984

Publication Date:
18 July 2018 (online)

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Summary

A series of hybridoma clones, each producing monoclonal antibodies to human tissue-type plasminogen activator (t-PA), were prepared from mice by standard procedures. Two of these clones were selected for further study. One HI72A1, produced antibodies that bound to t-PA and strongly inhibited its activity, whereas another, LI72D1, produced antibodies that bound to t-PA but did not affect its activity. The specificity of these antibodies was assessed in immunoabsorption experiments. Both immunoprecipitated 125I-labeled t-PA, and both were specific since only t-PA was recognized in conditioned media collected from Bowes melanoma cells cultured in the presence of 3H-leucine. Neither antibody recognized urokinase. t-PA was desorbed from antibody HI72A1-Sepharose columns with 0.5 M NaCl, consistent with its relatively low association constant (Ka = 9.37 × 107 M-1). In contrast, a strong chaotropic agent (i.e., 2 M KI) was required to elute t-PA from antibody LI72D1 columns (Ka = 2.08 × 109 M-1). This latter high affinity antibody was employed to develop an immunoradiometric assay for t-PA having a sensitivity of 0.5 ng/ml.

Keywords

Plasminogen activators - Monoclonal antibodies - Inhibition - Immunoradiometric assay
 

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