Academic Journal
Effective removal of host cell-derived nucleic acids bound to hepatitis B core antigen virus-like particles by heparin chromatography
| العنوان: | Effective removal of host cell-derived nucleic acids bound to hepatitis B core antigen virus-like particles by heparin chromatography |
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| المؤلفون: | Valentic, Angela, Hubbuch, Jürgen |
| المصدر: | Frontiers in Bioengineering and Biotechnology, 12, 1475918 ; ISSN: 2296-4185 |
| بيانات النشر: | Frontiers Media SA |
| سنة النشر: | 2024 |
| المجموعة: | KITopen (Karlsruhe Institute of Technologie) |
| مصطلحات موضوعية: | virus-like particles, heparin chromatography, HBcAg, host cell-derived nucleic acids, process development, ddc:660, Chemical engineering, info:eu-repo/classification/ddc/660 |
| الوصف: | Virus-like particles (VLPs) show considerable potential for a wide array of therapeutic applications, spanning from vaccines targeting infectious diseases to applications in cancer immunotherapy and drug delivery. In the context of hepatitis B core antigen (HBcAg) VLPs, a promising candidate for gene delivery approaches, the naturally occurring nucleic acid (NA) binding region is commonly utilized for effective binding of various types of therapeutic nucleic acids (NA$_{ther}$). During formation of the HBcAg VLPs, host cell-derived nucleic acids (NA$_{hc}$) might be associated to the NA binding region, and are thus encapsulated into the VLPs. Following a VLP harvest, the NA$_{hc}$ need to be removed effectively before loading the VLP with NA$_{ther}$. Various techniques reported in literature for this NA$_{hc}$ removal, including enzymatic treatments, alkaline treatment, and lithium chloride precipitation, lack quantitative evidence of sufficient NA$_{hc}$ removal accompanied by a subsequent high VLP protein recovery. In this study, we present a novel heparin chromatography-based process for effective NA$_{hc}$ removal from HBcAg VLPs. Six HBcAg VLP constructs with varying lengths of the NA binding region and diverse NA$_{hc}$ loadings were subjected to evaluation. Process performance was thoroughly examined through NA$_{hc}$ removal and VLP protein recovery analyses. Hereby, reversed phase chromatography combined with UV/Vis spectroscopy, as well as silica spin column-based chromatography coupled with dye-based fluorescence assay were employed. Additionally, alternative process variants, comprising sulfate chromatography and additional nuclease treatments, were investigated. Comparative analyses were conducted with LiCl precipitation and alkaline treatment procedures to ascertain the efficacy of the newly developed chromatography-based methods. Results revealed the superior performance of the heparin chromatography procedure in achieving high NA$_{hc}$ removal and concurrent VLP protein recovery. Furthermore, ... |
| نوع الوثيقة: | article in journal/newspaper |
| وصف الملف: | application/pdf |
| اللغة: | English |
| Relation: | info:eu-repo/semantics/altIdentifier/issn/2296-4185; https://publikationen.bibliothek.kit.edu/1000176407; https://publikationen.bibliothek.kit.edu/1000176407/155616552; https://doi.org/10.5445/IR/1000176407 |
| DOI: | 10.5445/IR/1000176407 |
| الاتاحة: | https://publikationen.bibliothek.kit.edu/1000176407 https://publikationen.bibliothek.kit.edu/1000176407/155616552 https://doi.org/10.5445/IR/1000176407 |
| Rights: | https://creativecommons.org/licenses/by/4.0/deed.de ; info:eu-repo/semantics/openAccess |
| رقم الانضمام: | edsbas.430B1537 |
| قاعدة البيانات: | BASE |
| DOI: | 10.5445/IR/1000176407 |
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