Size-selective purification of hepatitis B virus-like particle in flow-through chromatography: types of ion exchange adsorbent and grafted polymer architecture

Hon Wei Ng, Micky Fu Xiang Lee, Gek Kee Chua, Bee Koon Gan, Wen Siang Tan, Chien Wei Ooi, Siah Ying Tang, Eng Seng Chan, Beng Ti Tey

    Research output: Contribution to journalArticleResearchpeer-review

    Abstract

    Hepatitis B virus-like particles expressed in Escherichia coli were purified using anion exchange adsorbents grafted with polymer poly(oligo(ethylene glycol) methacrylate) in flow-through chromatography mode. The virus-like particles were selectively excluded, while the relatively smaller sized host cell proteins were absorbed. The exclusion of virus-like particles was governed by the accessibility of binding sites (the size of adsorbents and the charge of grafted dextran chains) as well as the architecture (branch-chain length) of the grafted polymer. The branch-chain length of grafted polymer was altered by changing the type of monomers used. The larger adsorbent (90 μm) had an approximately twofold increase in the flow-through recovery, as compared to the smaller adsorbent (30 μm). Generally, polymer-grafted adsorbents improved the exclusion of the virus-like particles. Overall, the middle branch-chain length polymer grafted on larger adsorbent showed optimal performance at 92% flow-through recovery with a purification factor of 1.53. A comparative study between the adsorbent with dextran grafts and the polymer-grafted adsorbent showed that a better exclusion of virus-like particles was achieved with the absorbent grafted with inert polymer. The grafted polymer was also shown to reduce strong interaction between binding sites and virus-like particles, which preserved the particles’ structure.

    Original languageEnglish
    Pages (from-to)2119-2129
    Number of pages11
    JournalJournal of Separation Science
    Volume41
    Issue number10
    DOIs
    Publication statusPublished - May 2018

    Keywords

    • adsorbents
    • anion exchange chromatography
    • flow-through chromatography
    • hepatitis B virus-like particles
    • size-selective adsorption

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