Method for the isolation of biological macromolecules and apparatus for conducting said method

摘要

Microtitration plates (14, 23) respectively including first parallel- and second cavities (16, 25) are arranged in apparatus (1, 41, 51) such that a first and a second cavity (16, 25) lie in succession. Filter units (20) are located in the first cavities (16), chromatographic material (29) in the second. The lysate is introduced into the first cavities. Solution leaving the first cavities is transferred directly into the second cavities, the macromolecules being adsorbed onto the chromatographic material. The endotoxins and other impurities are washed out. An elution fluid is introduced into the second cavities, so that mixed fluid and macromolecules leaves the second microtitration plate. The mixture is transferred to a third microtitration plate (32) with parallel third cavities (34). Ultrafiltration units (35) are located in the third cavities. The third cavities are subjected to a pressure difference or centrifugal force, acting towards their outlet openings. The macromolecules are recovered as retentate. The chromatographic material is equilibrated with a buffer, before solution delivery. After lysate addition to the first cavities, they are subjected to pressure difference or centrifugal force, acting towards the second cavities. This is supplemented by corresponding action on the second cavities. Further details of positioning and manipulation are elaborated. The agent used for washing is a nonionic tenside. This can be associated with an insert, a glass sinter or chromatographic material introduced. The filter units (20) are multistaged. The chromatographic material is an anion exchange material. It is located between two sinters; it is a monolithic block or a membrane. The washing agent impregnating the sinters, enriches endotoxins. The chromatographic material contains a washing agent enriching endotoxins. An Independent claim is included for corresponding apparatus.