Membrane based chromatographic or batch separation processes have been discussed as promising alternatives for some time now as they lead to more robust processes that are also very amenable to inclusion into a manufacturing process. While membranes provide a more easily accessible surface area for protein binding, they also help overcome the limitations of diffusion resistance, high pressure drop, need for high operating pressure/flow rate, and other usual problems related to traditional chromatographic process scale up. In our laboratory, regenerated cellulose membranes were linked to two different spacer arms (SA) and lysine residues to prepare affinity membranes for protein isolation.Thereafter, to compare their performance with that to other separation matrices, it is critical to know the lysine and SA density on membrane surface with high accuracy. The objective of this project is to modify published protocols that use Comassie brilliant blue (CBB) and 5(6)-carboxyfluorescein to quantify the L-lysine bound to the membrane; also to develop a Sodium Thiosulfate titration to determine the SA concentration bound on the membrane surface.Pieces of known dimensions were cut from the modified membranes and treated according to the modified protocols. The L-lysine and epoxy spacer arm densities on membrane surface were successfully determined and compared to that of commercially available matrices.

Analytical methods for the quantification of L-lysine on regenerated cellulose affinity membranes

247th National Meeting (2014)

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