Community Research and Development Information Service - CORDIS

A novel continuous affinity separation technique

The project involves the development of a process for separating pure proteins, such as enzymes or other products expressed by recombinant (genetically manipulated) organisms. The principle is based on the established procedure in which an anti-body which will react with the required protein is linked to a support (which may be packed in a column) producing an affinity material to which the sample material is brought into contact. The required protein binds onto the column, the impurities are washed off and the required protein is eluted in a pure form. This is a batch process. The purpose of this work is to replace this with a continuous method, binding the affinity material to a looped belt, which then passes through the crude sample, the washing process, a chamber where the required product is eluted and finally is regenerated. In spite of technical difficulties which initially limited yields and process rates, encouraging results were obtained, which should result in development of an effective apparatus.

The most successful of the experimental systems was that based on trypsin. This gave stable and reproducible results and was therefore used for much of the testing and development work. Continuous purifications of trypsin from pancreatic extract were carried out, the longest being of 30 h duration. The recovery was 30 40% at approximately 0.6 mg/h/1. Studies of the factors affecting the operation of the system showed that there was an inherent compromise between the efficiency of product recovery and the rate of purification. It was found that the problem of recovery were caused by the mixing of the chamber contents into a uniform concentration. A version of the apparatus was constructed that consisted of multiple subchambers for both the adsorption and elution steps of the process. which proved a success.


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