Preparing small affinity monoliths for antibody purification: Reproducibility of covalent immobilization of recombinant protein A on CIMmic™ CDI-0.1 column

CIMmic™ Monolithic Columns combine the advantages of the CIM® stationary phase with a flexible design and the possibility to operate with syringe. Discs containing the stationary phase can be easily interchanged inside the custom designed housing. Pre-activated chemistries enable immobilisation of numerous ligands and can be used for preparation of affinity chromatographic columns or enzyme reactors. Their small bed volume is particularly suitable for screening purposes and to optimise immobilisation protocols due to economic usage of often expensive ligands.

Carboxyimidazole (CDI) monolithic chromatographic columns are used for covalent immobilisation of proteins, peptides and other amine or thiol containing molecules. The covalent nature of the carbamate bond between the ligand and matrix reduces leaching and improves stability and reusability.

An example described below shows the feasibility of CIMmic™ CDI-0.1 utilisation for covalent immobilisation of recombinant protein A (r-pA). Additionally the example was used for the evaluation of the reproducibility of CIMmic™ CDI columns. The dynamic binding capacity for human polyclonal immunoglobulin (IgG) was used as metric for comparison of the affinity columns.