Purification of protein inclusion bodies by crossflow microfiltration
View Patent ↗The invention relates to a process for purifying and/or concentrating inclusion bodies (refractile bodies) in a highly concentrated particle-containing solution, in which process the inclusion body-containing suspension is directed tangentially past one or more semipermeable membranes, so that the inclusion bodies are retained behind the membranes and that substances having a relatively small molecular weight may pass through the membrane and/or be adsorbed to the membrane, and a purified and/or concentrated inclusion body suspension is obtained. In the same manner, cell wall particles are removed from a inclusion body suspension. The invention further relates to the use of a crossflow microfiltration unit for purifying and/or concentrating inclusion bodies in a suspension and for removing cell wall particles from a inclusion body preparation.
1. A process for purifying and/or concentrating a biomass load of inclusion bodies in a solution, which is characterized in that the inclusion body-containing solution is directed tangentially past one or more semipermeable membranes so that the membranes retain the inclusion bodies and that substances or components having a relatively small molecular weight or particle diameter can pass through, thus obtaining a purified and/or concentrated inclusion body solution, and on the semipermeable membrane a planar net matrix is located which is constructed of intersecting longitudinal and transverse filaments such that the adjacent longitudinal and transverse filaments are, in each case, at a distance from one another which is 5 to 15 times greater than their thickness which is in the range from 150 to 600 μm, said matrix having a membrane to matrix distance of about 80 μm, a matrix thickness of about 760 μm, a filament thickness of about 340-450 μm and a filament distance of about 2400 μm.
2. The process according to claim 1 , characterized in that the biomass load of the membrane is greater than 65 g of wet cell mass equivalent/m 2 .
3. The process according to claim 1 , characterized in that the biomass load of the membrane is greater than 650 g of wet cell mass equivalent/m 2 .