Lipid foulant interactions during the chromatographic purification of virus-like particles from Saccharomyces cerevisiae

摘要: The objective of this study was to understand the mechanism lipid fouling in chromatography through investigation a hydrophobic interaction chromatography (HIC) operation. This motivated by need phenomenon during manufacture biological products such as vaccines. The systematic approach and novel analytical techniques employed create unique platform other chromatographic adsorbents process feed materials. HIC is primary capture step purification yeast derived hepatitis R surface antigen (HBsAg), where required cell disruption detergent liberation steps release high levels content into feed stream. From lipid- rich lipid-depleted feedstocks, comparative analysis able quantify deterioration performance (binding capacities, purities recoveries) under successive cycles. Furthermore, full mass balance on host lipids identified highly triacylglyceride main foulant. Intra-particle distribution and progression its effects material adsorption diffusion were then examined under confocal laser scanning microscopy (CLSM). In addition, high- resolution electron (SEM) images fouled bead (after 40 cycles) confirmed that thick layer building up outer surface. Based these findings, thought be rapid accumulation foulant at rim bead, which aggravated the possible hindrance resulting from multi adsorption. Finally, pretreatments reduce evaluated terms improvement quality performance. Selective adsorbent polystyrene XAD-4 demonstrated promising removal capabilities with satisfactory HBsAg VLP recoveries. improved column resulted a three-fold increase product capacity, whilst overall yield remained constant over