Preparation of pure human tumor necrosis factor and hybridomas producing monoclonal antibodies to human tumor necrosis factor

US4870163A · kind A · utility

Assignees

• New York Blood Center, Inc. · US
• Sloan Kettering Institute for Cancer Research · US

Inventors

• Berish Y. Rubin · Brooklyn, US
• Sylvia L. Anderson · Cresskill, US
• Susan A. Sullivan · Brooklyn, US
• Lloyd J. Old · New York, US
• Barbara Williamson · Stamford, US
• Elizabeth C. Richards · Tarrytown, US

Key dates

• Filing date: Aug 29, 1985
• Grant date: Sep 26, 1989
• Expiry date: Aug 29, 2005

Classification

• Technology area (CPC Y): Emerging Cross-Sectional Technologies
• CPC primary: Y10S530/829
• WIPO field: Biotechnology
• WIPO sector: Chemistry

Abstract

Human TNF(LuKII) having a specific activity of at least 1.5.times.10.sup.5 units per milligram of total protein is produced by contacting a TNF-containing protein composition, which has been harvested from human cell lines of hematopoietic origin or recombinant origin, in separate adsorption stages with glass beads, lentil lectin bound to Sepharose, and procion red agarose, thereby selectively to adsorb TNF in each stage, while leaving some impurities unadsorbed, each contact stage being followed by contact of the adsorbent with an eluant thereby to obtain a solution of more highly purified TNF after each stage. The purified human TNF(LuKII) is used to produce monoclonal antibodies against TNF(LuKII) and such antibodies can be used to assay samples for the presence of TNF(LuKII).