Monoclonal antibodies against a recombinant form of plasminogen activator inhibitor-1: Effects on tissue plasminogen activator neutralizing and vitronectin binding properties |
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| Affiliation: | 1. Department of Structural Biology, Weizmann Institute of Science, Rehovot 7610001, Israel;2. Department of Chemical Research Support, Weizmann Institute of Science, Rehovot 7610001, Israel;3. De Botton Institute for Protein Profiling, Nancy and Stephen Grand Israel National Center for Personalized Medicine, Weizmann Institute of Science, Rehovot 7610001, Israel;1. Medical Research Institute of New Zealand, Wellington, New Zealand;2. Capital and Coast District Health Board, Wellington, New Zealand;3. Division of Respiratory Medicine, School of Clinical Sciences, University of Nottingham, Nottingham, United Kingdom;4. Clinical Management Group, Woolcock Institute of Medical Research, University of Sydney, Sydney, Australia;5. Henderson Medical Centre, Auckland, New Zealand;6. Pharmacology & Clinical Pharmacology, University of Auckland, Auckland, New Zealand;7. School of Medicine, University of Otago, Wellington, New Zealand;1. National Institute for Biological Standards and Control (NIBSC), Blanche Lane, South Mimms, Potters Bar, Hertfordshire, UK;2. Amgen Inc., 1120 Veterans Blvd, South San Francisco CA 94080, USA;3. Drug Safety Research and Development, Pfizer, Inc., Groton, CT 06340, USA;4. Roche Innovation Center, Basel, Switzerland. Pharmaceutical Sciences Switzerland;5. GlaxoSmithKline, 1250 South Collegeville Road, Collegeville, PA 19426, USA;6. Bristol-Myers Squibb, 10300 Campus Point Drive, Suite 100, San Diego, CA 92121, USA;7. Genentech, 1 DNA Way, South San Francisco, CA 94080, USA;8. Oncology Safety, Clinical Pharmacology & Safety Sciences, R&D, AstraZeneca, Cambridge, UK;9. Janssen R&D, 1400 McKean Road, Spring House, PA 19477, USA;10. MRL, Merck & Co., Inc., 213 E Grand Ave, South San Francisco, CA 94080, USA;11. Novartis Institutes for BioMedical Research, Klybeckstrasse 141, Basel CH-4002, Switzerland;1. Genoval Therapeutics Co., Ltd, Shanghai, China;2. Cardiometabolic Center, State Key Laboratory of Cardiovascular Disease, FuWai Hospital, National Center for Cardiovascular Diseases, National Clinical Research Center for Cardiovascular Diseases, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, China;3. Heart Failure Center, State Key Laboratory of Cardiovascular Disease, FuWai Hospital, National Center for Cardiovascular Diseases, National Clinical Research Center for Cardiovascular Diseases, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, China |
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| Abstract: | A panel of eight murine monoclonal antibodies was produced against a recombinant form of plasminogen activator inhibitor- 1 (rPAI-1). All the antibodies recognized active and latent forms of rPAI-1, and rPAI-1 complexed with tissue plasminogen activator (t-PA) as determined in enzyme-linked immunosorbent assays (ELISA). Three of the antibodies, FAG9, FAD3 and BBH2, were particularly effective at inhibiting the t-PA neutralizing activity of rPAI-1 as measured in a chromogenic assay. Three different antibodies, DD8E9, FEG7 and FGG7, inhibited the binding of [125I]rPAI-1 to vitronectin immobilized on polystyrene wells. The combination of FGG7 and alkaline phosphatase-labeled BBH2 resulted in a sensitive sandwich ELISA for rPAI-I with detection limits in the range of 1–10 ng. Definition of epitopes recognized by these antibodies will be useful for identifying various domains on PAI-1 involved in its interaction with protease substrates and with its protein cofactor, vitronectin. |
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