دورية أكاديمية
Comprehensive Comparison of AAV Purification Methods: Iodixanol Gradient Centrifugation vs. Immuno-Affinity Chromatography.
| العنوان: | Comprehensive Comparison of AAV Purification Methods: Iodixanol Gradient Centrifugation vs. Immuno-Affinity Chromatography. |
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| المؤلفون: | Lam AK; Department of Pediatrics, Herman B Wells Center for Pediatric Research, Indiana University School of Medicine, Indianapolis, IN 46202, USA., Mulcrone PL; Department of Pediatrics, Herman B Wells Center for Pediatric Research, Indiana University School of Medicine, Indianapolis, IN 46202, USA., Frabutt D; Department of Pediatrics, Herman B Wells Center for Pediatric Research, Indiana University School of Medicine, Indianapolis, IN 46202, USA., Zhang J; Department of Pediatrics, Herman B Wells Center for Pediatric Research, Indiana University School of Medicine, Indianapolis, IN 46202, USA., Chrzanowski M; Lewis Katz School of Medicine, Temple University, Philadelphia, PA 19140, USA., Arisa S; Department of Pediatrics, Herman B Wells Center for Pediatric Research, Indiana University School of Medicine, Indianapolis, IN 46202, USA., Munoz M; Department of Pediatrics, Herman B Wells Center for Pediatric Research, Indiana University School of Medicine, Indianapolis, IN 46202, USA., Li X; Department of Pediatrics, Herman B Wells Center for Pediatric Research, Indiana University School of Medicine, Indianapolis, IN 46202, USA., Biswas M; Department of Pediatrics, Herman B Wells Center for Pediatric Research, Indiana University School of Medicine, Indianapolis, IN 46202, USA., Markusic D; Department of Pediatrics, Herman B Wells Center for Pediatric Research, Indiana University School of Medicine, Indianapolis, IN 46202, USA., Herzog RW; Department of Pediatrics, Herman B Wells Center for Pediatric Research, Indiana University School of Medicine, Indianapolis, IN 46202, USA., Xiao W; Department of Pediatrics, Herman B Wells Center for Pediatric Research, Indiana University School of Medicine, Indianapolis, IN 46202, USA. |
| المصدر: | Advances in cell and gene therapy [Adv Cell Gene Ther] 2023; Vol. 2023. Date of Electronic Publication: 2023 Dec 11. |
| نوع المنشور: | Journal Article |
| اللغة: | English |
| بيانات الدورية: | Publisher: Wiley Country of Publication: United States NLM ID: 101731783 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 2573-8461 (Electronic) Linking ISSN: 25738461 NLM ISO Abbreviation: Adv Cell Gene Ther Subsets: PubMed not MEDLINE |
| أسماء مطبوعة: | Original Publication: Hoboken, NJ : Wiley |
| مستخلص: | Recombinant adeno-associated viruses (AAVs) have emerged as a widely used gene delivery platform for both basic research and human gene therapy. To ensure and improve the safety profile of AAV vectors, substantial efforts have been dedicated to the vector production process development using suspension HEK293 cells. Here, we studied and compared two downstream purification methods, iodixanol gradient ultracentrifugation versus immuno-affinity chromatography (POROS ™ CaptureSelect ™ AAVX column). We tested multiple vector batches that were separately produced (including AAV5, AAV8, and AAV9 serotypes). To account for batch-to-batch variability, each batch was halved for subsequent purification by either iodixanol gradient centrifugation or affinity chromatography. In parallel, purified vectors were characterized, and transduction was compared both in vitro and in vivo in mice (using multiple transgenes: Gaussia luciferase, eGFP, and human factor IX). Each purification method was found to have its own advantages and disadvantages regarding purity, viral genome (vg) recovery, and relative empty particle content. Differences in transduction efficiency were found to reflect batch-to-batch variability rather than disparities between the two purification methods, which were similarly capable of yielding potent AAV vectors. Competing Interests: Conflicts of Interest The authors declare that they have no conflicts of interest. |
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| معلومات مُعتمدة: | P01 HL160472 United States HL NHLBI NIH HHS; R01 HL130871 United States HL NHLBI NIH HHS; T32 HL007910 United States HL NHLBI NIH HHS; U54 HL142019 United States HL NHLBI NIH HHS |
| تواريخ الأحداث: | Date Created: 20231222 Latest Revision: 20240210 |
| رمز التحديث: | 20240210 |
| مُعرف محوري في PubMed: | PMC10735247 |
| DOI: | 10.1155/2023/2339702 |
| PMID: | 38130431 |
| قاعدة البيانات: | MEDLINE |
| تدمد: | 2573-8461 |
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| DOI: | 10.1155/2023/2339702 |