Bio-Works resins, with high binding capacities and CIP using NaOH up to 0.5 M, are ideal for purifying antibodies
The expression levels of monoclonal antibodies (mAb) in biopharma production are constantly increasing and are now frequently up to 10 g/L. Efficient production therefore often requires a capture step using chromatography resins with high binding capacities over a short time and with efficient cleaning-in-place stability using NaOH up to 0.5 M. Protein A resins give in general purities of >85-90% using standard conditions without optimization, which often is enough for many lab scale purifications.
Demanding large-scale purification usually requires optimization of the washing and elution steps followed by one or more polishing purification steps using e.g. ion-exchange chromatography.
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WorkBeads Protein A
WorkBeads Protein A is an agarose-based resin with high binding capacity and with a cleaning-in-place stability up to 10 mM NaOH. The protein A ligand is coupled to the resin using a bromohydrin-based method that gives high chemical stability and low ligand leakage. High capacity and a well-established agarose matrix make WorkBeads Protein A ideal for purification of monoclonal antibodies (mAb) as well as polyclonal antibodies. GoBio Mini A 1 ml and GoBio Mini A 5 ml are prepacked columns with WorkBeads Protein A.
WorkBeads 40 TREN
Purification of antibodies from mammalian host cells, such as Chinese Hamster Ovary (CHO), results in extensive bioburden on the protein A column. Chromatins, together with host cell proteins in general, cause damage to the protein A column. Regular cleaning-in-place (CIP) is mandatory in the purification process, but accumulative fouling of the column will still occur. Maximized lifetime of the protein A column is thus an important requirement. It would be desirable to add a purification step before the protein A purification step to reduce the components that constitute the worst threat, and in the process improve the final purity of the target antibody.
WorkBeads 40 TREN is a multimodal ion exchange chromatography (IEX) resin with a ligand (tris(2-aminoethyl)amine, TAEA) that is positively charged below pH 9. The resin has been demonstrated to be effective in removal of impurities such as chromatin complexes (nucleic acid-protein complexes), endotoxins and host cell proteins. We introduced WorkBeads 40 TREN upstream of WorkBeads affimAb in the flow scheme and used it in flow through mode to adsorb undesirable impurities while letting mAb pass through the column without binding.