PRODUCTION OF HUMAN INTERLEUKIN-8 EXPRESSED IN ESCHERICHIA-COLI - FROM A LABORATORY-SCALE FOR IN-VITRO TESTS VIA A TECHNICAL SCALE FOR ANIMAL STUDIES TO A PROCESS-SCALE FOR A GMP-COMPATIBLE PRODUCTION

An Escherichia coil K 12 strain has been constructed for efficient exp ression of recombinant biologically active human IL-8 (Interleukin-8). The development of a fermentation and purification process from the l aboratory scale (cells from 15 1 fermentation broth) to a production s cale (cells from 200 1 fermentation broth) is described. Material obta ined from the laboratory scale was used for initial in vitro studies a nd for the development of a biological assay. An upscale purification process starting from 80 I fermentation broth resulted in larger amoun ts of IL-8 needed for preclinical studies. This process includes a ful ly automated control of the initial affinity chromatography step. Fina lly, a production process which differed markedly from the small-scale processes was tailor-made for GMP conformity and economic considerati ons. It consists of a cell disruption step followed by two crossflow d iafiltrations with different molecular weight cut offs and filtration rates, one cation exchange chromatography and a final dialysis step. I n order to enhance the overall yield of biologically active IL-8, cond itions for a resolubilisation of insoluble IL-8 present in the remaini ng pellet after cell disruption were worked out. (C) 1997 Elsevier Sci ence B.V.