La Jolla Cancer Research Foundation

The Salk Institute Biotechnology Industrial Associates, Inc.

Thomas S Vedvick

William S Craig

John R Harper

Jonathan R Parker

Paul J Kostel

Richard G Buckholz

Jennifer A Kinney

Sam D Hernandez

Mary S Urcan

Michael E Engel

The invention provides a process for detecting the presence of guanidinium ions in a sample solution. The detection process involves contacting a sample solution suspected of containing guanidinium ions with a cation exchange resin and eluting the guanidinium ions present in the sample solution with an aqueous buffer solution having a pH of about 1.5 to about 2. This is followed by contacting the eluant with a cation suppressor column and simultaneously flowing a suppressor regenerate solution in the opposite direction on the opposite side of the permeable membrane of the column, and finally, detecting the presence of guanidinium ions in the eluant from the ion exchange column which was contacted with the suppressor column by use of a conductivity detector.

Processes for the purification of human recombinant decorin and the

The present invention is directed to a process for the production of substantially pure human recombinant decorin which involves the combination of three separate stages characterized by contacting decorin-containing cell culture medium with (1) a first strong anionic exchange resin; then with (2) a hydrophobic interactive chromatographic resin; and finally with (3) a second strong anionic exchange resin. By using a combination of steps, and certain reagents, in particular a 2.4 to 3 molar GuHCl solution to elute decorin from a hydrophobic interactive column, the process of this invention provides a more convenient and reproducible process for purifying human recombinant decorin. The invention also provides a process for detecting the presence of guanidinium ions in a sample solution. The detection process involves contacting a sample solution suspected of containing guanidinium ions with a cation exchange resin and eluting the guanidinium ions present in tire sample solution with an aqueous buffer solution having a pH of about 1.5 to about 2. This is followed by contacting the eluant with a cation suppressor columns and simultaneously flowing a suppressor regenerate solution in the opposite direction on the opposite side of the permeable membrane of the column, and finally, detecting the presence of guanidinium ions in the eluant from the ion exchange column which was contacted with the suppressor column by use of a conductivity detector.

Biologically active aprotinin (APR) molecules, naturally occurring, relatively short, single chain polypeptides, are prepared by growing methylotrophic yeast transformants containing in their genome at least one copy of a DNA sequence operably encoding APR, in operational association with a DNA sequence encoding the S. cerevisiae alpha mating factor pre-pro sequence (including a processing sequence selected from lys-arg-(glu-ala).sub.x, wherein x is an integer falling in the range of 0-3), both under the regulation of a promoter region of a gene of a methylotrophic yeast, under conditions allowing expression of said DNA sequences, and secretion of APR molecules into the culture medium. Also disclosed are novel DNA fragments and novel recombinant yeast strains which are useful in the practice of the present invention.

DNA for expression of aprotinin in methylotrophic yeast cells

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Carlsbad, CA, United States of America

Thomas S Vedvick