‘Next generation’ chromatographic media for biopharmaceutical manufacturing

[摘要] Novel ways to produce bilayered chromatography adsorbent media featuring a functionalised core surrounded by an inert size exclusion layer for nanoplex purification were explored. Surface functionalisation of anion exchange chromatography media via cold atmospheric pressure plasma etching and oxidation was investigated. Both, a dielectric barrier discharge plasma generation system and a fluidised bed underwater reactor produced bead samples with significantly reduced surface binding, while maintaining their core binding capacities. A 3D visualisation method, allowing the study of the binding topography of the spatial distribution of pDNA and BSA adhered to Q ligands within particles was developed. This CLSM method revealed imperfections on the surface of adsorbents, providing additional binding sites for pDNA due to micro crevices. Implementing beads with a more homogenous surface, restricted access media were produced via an AGE activation-partial bromination method. The thickness and homogeneity of the size exclusion layer was controlled via viscosity enhanced reaction diffusion balancing, yielding a distinct layer devoid of ligands. The particles did not show residual pDNA binding and selectivity of core vs surface binding increased by 100-fold cf. previous studies. The commercial adsorbent Capto Core 700 was successfully implemented as a first capture step for pDNA purification from crude E.coli lysates.

[发布机构] University:University of Birmingham;Department:School of Chemical Engineering

[关键词] Q Science;Q Science (General)