TY - JOUR
T1 - Epitope mapping of monoclonal antibodies directed to PAI-1 using PAI-1/PAI-2 chimera and PAI-1-derived synthetic peptides
AU - Muehlenweg, Bernd
AU - Guthaus, Elke
AU - De Prada, Nuria Arroyo
AU - Schmitt, Manfred
AU - Schmiedeberg, Niko
AU - Kotzsch, Matthias
AU - Creutzburg, Sabine
AU - Kramer, Michael D.
AU - Kessler, Horst
AU - Wilhelm, Olaf G.
AU - Magdolen, Viktor
N1 - Funding Information:
We thank P.A. Andreasen (Department of Molecular and Structural Biology, Aarhus University, Denmark) for providing mAbs to PAI-1 and helpful discussions, C. Weiss (Department of Orthopedic Surgery, TU München) for initial work on generating PAI-1 and PAI-2 expression plasmids, and T. Wöhl (Wilex Biotechnology GmbH, München) for serpin alignments. Part of this work was supported by grants of the Sonderforschungsbereich 469 of the Ludwig-Maximilians-Universität München, of the Graduiertenkolleg 333 of the Deutsche Forschungsgemeinschaft and of the Deutsche Krebshilfe e.V. (Dr. Mildred Scheel-Stiftung).
PY - 2000/4/1
Y1 - 2000/4/1
N2 - Plasminogen activator inhibitor type-1 is a key regulatory protein of the fibrinolytic system that is involved in a variety of physiological and pathophysiological processes. A panel of 14 monoclonal antibodies directed against plasminogen activator inhibitor type-1 was analyzed regarding epitope specificity on plasminogen activator inhibitor type-1. For this purpose, chimera consisting of plasminogen activator inhibitor type-1 and another plasminogen activator inhibitor, plasminogen activator inhibitor type-2, with different portions of the respective wild-type proteins, were generated and plasminogen activator inhibitor type-1-derived 20-mer and 10-mer linear peptides were synthesized. Nine of the 14 monoclonal antibodies recognized an epitope located in the region between amino acid 76-188 of plasminogen activator inhibitor type-1, which encompasses the binding sites for vitronectin, heparin, and part of the fibrin binding region. Of these nine monoclonal antibodies, six reacted with a quadruple plasminogen activator inhibitor type-1 mutant (N152H, K156T, Q321L, M356I), and seven detected a plasminogen activator inhibitor type-1 deletion mutant (ΔF111-H114). Two of the remaining five monoclonal antibodies recognized epitopes located between amino acid 209-227 and amino acid 352-371, respectively, while the other three antibodies reacted with wild-type plasminogen activator inhibitor type-1, only. Additional experiments revealed that two of the 14 mAbs neutralized and one monoclonal antibodies increased plasminogen activator inhibitor type-1 activity toward urokinase-type plasminogen activator, one of its target proteases. Copyright (C) 2000 Elsevier Science Ltd.
AB - Plasminogen activator inhibitor type-1 is a key regulatory protein of the fibrinolytic system that is involved in a variety of physiological and pathophysiological processes. A panel of 14 monoclonal antibodies directed against plasminogen activator inhibitor type-1 was analyzed regarding epitope specificity on plasminogen activator inhibitor type-1. For this purpose, chimera consisting of plasminogen activator inhibitor type-1 and another plasminogen activator inhibitor, plasminogen activator inhibitor type-2, with different portions of the respective wild-type proteins, were generated and plasminogen activator inhibitor type-1-derived 20-mer and 10-mer linear peptides were synthesized. Nine of the 14 monoclonal antibodies recognized an epitope located in the region between amino acid 76-188 of plasminogen activator inhibitor type-1, which encompasses the binding sites for vitronectin, heparin, and part of the fibrin binding region. Of these nine monoclonal antibodies, six reacted with a quadruple plasminogen activator inhibitor type-1 mutant (N152H, K156T, Q321L, M356I), and seven detected a plasminogen activator inhibitor type-1 deletion mutant (ΔF111-H114). Two of the remaining five monoclonal antibodies recognized epitopes located between amino acid 209-227 and amino acid 352-371, respectively, while the other three antibodies reacted with wild-type plasminogen activator inhibitor type-1, only. Additional experiments revealed that two of the 14 mAbs neutralized and one monoclonal antibodies increased plasminogen activator inhibitor type-1 activity toward urokinase-type plasminogen activator, one of its target proteases. Copyright (C) 2000 Elsevier Science Ltd.
KW - Chimera
KW - Monoclonal antibodies
KW - Plasminogen activator inhibitor type-1
KW - Synthetic peptides
UR - http://www.scopus.com/inward/record.url?scp=0033997385&partnerID=8YFLogxK
U2 - 10.1016/S0049-3848(99)00238-8
DO - 10.1016/S0049-3848(99)00238-8
M3 - Article
C2 - 10706935
AN - SCOPUS:0033997385
SN - 0049-3848
VL - 98
SP - 73
EP - 81
JO - Thrombosis Research
JF - Thrombosis Research
IS - 1
ER -