Rosario R Martinez Garcia
The Center for Molecular Immunology, Cuba
Title: Performance of the AbSolute® High Cap resin in the Nimotuzumab capture step
Biography
Biography: Rosario R Martinez Garcia
Abstract
Protein A is the affinity chromatography ligand of choice for first-step capture in the purification of mAbs as its high selectivity gives excellent purity plus high yields and also functions as the key volume reduction step in the process since Protein A concentrates the product stream. The relatively high cost of these resins leads to consider different operational strategy as evaluate protein A matrixes with better performance in dynamic binding capacity at high flow rates. Recently a new Protein A ligand, AbSolute® High Cap (AbSolute HC), was introduced by Novasep Company that is modified with respect to high binding capacity at higher velocities. This study investigated the performance of AbSolute HC resin during Nimotuzumab capture by measuring its DBC at different flow rates and feed concentrations at lab scale. AbSolute HC data were also compared with Mab Select SuRe, current affinity gel used in the purification process. The process conditions were adjusted including the maximum speed for all steps with an assessment of impact on Nimotuzumab purity using AbSolute HC. In addition the impact of loading amount on aggregates formation with this media and the resin lifetime were evaluated. The results showed better DBC at 10% on AbSolute HC than Mab Select Sure, with a higher DBC at 10% on Absolute HC with IgG concentration at 2 g/L than 0.13 g/L. No significant loss of purity or yield for speeds until 1000 cm/h for all steps was obtained using AbSolute HC resin. The lifetime of the media was tested up to 200 cycles with suitably results using NaOH and PAB solution.