University of California

Pharmacia & Upjohn Company

Novartis Vaccines & Diagnostics, Gmbh

Geomatrix Consultants, Inc.

Douglas G Bolesch

Jay D Keasling

Glenn J Dorin

Cynthia A Cowgill

Patricio T Riquelme

Mark E Gustafson

David H Reifsnyder

Thomas A Delfino

Duane Inlow

Highly purified preparations of TFPI or TFPI analogs can be prepared using a method that generally involves the following steps: (1) expression of TFPI or TFPI analog in, (2) isolation of refractile bodies, (3) dissolution of the refractile bodies and refolding of the expressed TFPI or TFPI analog, (4) SP-Sepharose fast flow (FF) chromatography, (5) a first concentration and diafiltration step, (6) Q-Sepharose high (HP) performance chromatography, (7) butyl hydrophobic interaction chromatography (HIC), (8) SP-Sepharose HP chromatography, and (9) a second concentration/diafiltration step. Less than about 12% of the TFPI or TFPI analog molecules in such preparations are modified TFPI or TFPI analog species (i.e., oxidized, carbamylated, acetylated, deamidated, aggregated, or misfolded species).

Method of purifying TFPI and TFPI analogs

The invention provides methods and microbial cultures for the bioremediation of organic halide contaminated groundwater contaminated with organic halides, such as di- and trichloroethene. The methods involve adding, in situ to organic halide-contaminated groundwater a carbohydrate and one or more reductive dehalogenation factors, usually in the form of a nutrient extract, both in amounts sufficient to permit in situ reductive dehalogenation of the organic halide by a microbial population. The microbial population may be endogenous to the ground water or added exogenously. The nutrient-enriched ground water is then maintained in situ under reducing conditions to reductively dehalogenate the contaminating organic halide. Enriched bioremediation cultures are produced by adding to organic halide contaminated groundwater which comprises an endogenous microbial population capable of reductive dehalogenation of the organic halide a carbohydrate and frequently, one or more reductive dehalogenation factors. Thereafter, the nutrient-enriched groundwater is incubated under reducing conditions whereby the organic halide is reductively dehalogenated and the microbial population is selectively and numerically expanded to yield a bioremediation culture.

Reductive dehalogenation of organic halides in contaminated groundwater

Growing community of inventors

Berkeley, CA, United States of America

Douglas G Bolesch