TY - JOUR
T1 - Negative chromatography of hepatitis B virus-like particle
T2 - comparative study of different adsorbent designs
AU - Lee, Micky Fu Xiang
AU - Chan, Eng Seng
AU - Tan, Wen Siang
AU - Tam, Kam Chiu
AU - Tey, Beng Ti
N1 - Funding Information:
MFX Lee is currently supported by the HDR Scholarship from School of Engineering, Monash University Malaysia. The study is funded by e-Science Fund (06-02-10-SF0255) from the Ministry of Science, Technology and Innovation, Malaysia. We acknowledge BioToolomics Ltd. for providing InertShell and InertLayer 1000.
Funding Information:
MFX Lee is currently supported by the HDR Scholarship from School of Engineering, Monash University Malaysia. The study is funded by e-Science Fund ( 06-02-10-SF0255 ) from the Ministry of Science, Technology and Innovation, Malaysia. We acknowledge BioToolomics Ltd. for providing InertShell and InertLayer 1000.
Publisher Copyright:
© 2016 Elsevier B.V.
Copyright:
Copyright 2020 Elsevier B.V., All rights reserved.
PY - 2016/5/6
Y1 - 2016/5/6
N2 - Purification of virus-like particles (VLPs) in bind-and-elute mode has reached a bottleneck. Negative chromatography has emerged as the alternative solution; however, benchmark of negative chromatography media and their respective optimized conditions are absent. Hence, this study was carried out to compare the performance of different negative chromatography media for the purification of hepatitis B VLPs (HB-VLPs) from clarified Escherichia coli feedstock. The modified anion exchange media, core-shell adsorbents (InertShell and InertLayer 1000) and polymer grafted adsorbents (SQ) were compared. The results of chromatography from packed bed column of core-shell adsorbents showed that there is a trade-off between the purity and recovery of HB-VLPs in the flowthrough fraction due to the shell thickness. Atomic force microscopic analysis revealed funnel-shaped pore channels in the shell layer which may contribute to the entrapment of HB-VLPs. A longer residence time at a lower feed flow rate (0.5 ml/min) improved slightly the HB-VLPs purity in all modified adsorbents, but the recovery in InertShell reduced substantially. The preheat-treatment is not recommended for the negative chromatography as the thermal-induced co-aggregation of HCPs and HB-VLPs would flow along with HB-VLPs and thus reduced the HB-VLPs purity in the flowthrough. Further reduction in the feedstock concentration enhanced the purity of HB-VLPs especially in InertLayer 1000 but reduced substantially the recovery of HB-VLPs. In general, the polymer grafted adsorbent, SQ, performed better than the core-shell adsorbents in handling a higher feedstock concentration.
AB - Purification of virus-like particles (VLPs) in bind-and-elute mode has reached a bottleneck. Negative chromatography has emerged as the alternative solution; however, benchmark of negative chromatography media and their respective optimized conditions are absent. Hence, this study was carried out to compare the performance of different negative chromatography media for the purification of hepatitis B VLPs (HB-VLPs) from clarified Escherichia coli feedstock. The modified anion exchange media, core-shell adsorbents (InertShell and InertLayer 1000) and polymer grafted adsorbents (SQ) were compared. The results of chromatography from packed bed column of core-shell adsorbents showed that there is a trade-off between the purity and recovery of HB-VLPs in the flowthrough fraction due to the shell thickness. Atomic force microscopic analysis revealed funnel-shaped pore channels in the shell layer which may contribute to the entrapment of HB-VLPs. A longer residence time at a lower feed flow rate (0.5 ml/min) improved slightly the HB-VLPs purity in all modified adsorbents, but the recovery in InertShell reduced substantially. The preheat-treatment is not recommended for the negative chromatography as the thermal-induced co-aggregation of HCPs and HB-VLPs would flow along with HB-VLPs and thus reduced the HB-VLPs purity in the flowthrough. Further reduction in the feedstock concentration enhanced the purity of HB-VLPs especially in InertLayer 1000 but reduced substantially the recovery of HB-VLPs. In general, the polymer grafted adsorbent, SQ, performed better than the core-shell adsorbents in handling a higher feedstock concentration.
KW - Core-shell adsorbents
KW - Negative chromatography
KW - Poly[oligo(ethylene glycol)methacrylate]
KW - Preheat-treatment
KW - Virus-like particles
UR - http://www.scopus.com/inward/record.url?scp=84979468725&partnerID=8YFLogxK
U2 - 10.1016/j.chroma.2016.03.066
DO - 10.1016/j.chroma.2016.03.066
M3 - Article
C2 - 27059397
AN - SCOPUS:84979468725
SN - 0021-9673
VL - 1445
SP - 1
EP - 9
JO - Journal of Chromatography A
JF - Journal of Chromatography A
ER -