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A Rapid Procedure for the Large Scale Purification of Elastase and Cathepsin G From Human Sputum

A Rapid Procedure for the Large Scale Purification of Elastase and Cathepsin G From Human Sputum

R R Martodam, R J Baugh, D Y Twumasi, I E Liener

Prep Biochem. 1979;9(1):15-31.

PMID: 441048

Abstract:

A procedure is described which permits the rapid isolation of large amounts of elastase and cathepsin G from purulent sputum. This procedure involves: (1) digestion of sputum with DNase, (2) extraction of the insoluble residue that remains with 1 M NaCl, pH 8, (3) affinity chromatography on Sepharose-bound Trasylol, and (4) separation of the two enzymes by chromatogrphy on CM-Sephadex. Starting with 500 g of sputum it was possible to isolate 175 mg of each of these two enzymes within 7 to 10 days. Active site titration indicated both enzymes to be at least 97% pure. Disc gel electrophoresis in the presence and absence of SDS and amino acid sequence of the N-terminal region support the conclusion that the elastase and cathepsin G isolated from sputum are identical to the same enzymes isolated directly from the leukocytes of human blood.

Chemicals Related in the Paper:

Catalog Number	Product Name	Structure	CAS Number	Price
AP107200920	Cathepsin G from human sputum		107200-92-0	Price

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