Batch Chromatography for Protein Concentration

Summary

Batch chromatography is simple and fast, suitable for situations where protein resolution is not required, and an effective means of concentrating proteins. Source: Handbook of Protein Technology

Operation method

batch chromatography

Principle

Batch chromatography is characterized by the fact that the resin for chromatography is not placed in a column but in a container of beakers or bottles; the in-mixed chromatography is performed by stirring or shaking; and the chromatant is separated by filtration or centrifugation.

Materials and Instruments

Protein solutions
Ion exchange chromatography device

Move

1. equilibrate the resin in a beaker or conical flask, stirring constantly to promote equilibration;

2. pour off most of the solution, reserving the solution so that the resin can be easily stirred;

3. Add sample;

4. Allow to stand for 1 h to allow the proteins to fully bind to the resin, stirring several times in between. 5;

5. filter the resin, taking care not to dehydrate the resin;

6. gently rinse the resin with the original solution on the filter. 7. put the resin back into the beaker;

7. put the resin back into the beaker and treat it with a solution of high ionic strength. 8. filter the resin and check the elution;

8. filtering the resin and testing the eluate for the presence of proteins

9. if necessary, treat the resin with a solution of higher ionic strength until the target protein is washed out.

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