Preparation of pure human tumor necrosis factor and hybridomas p

Chemistry: natural resins or derivatives; peptides or proteins; – Proteins – i.e. – more than 100 amino acid residues – Separation or purification

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530395, 530415, 530829, 435 7, 4351722, 43524027, 436548, C07K 312, C07K 318, C07K 320

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048701637

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).

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