Methodological Study on Determination of Recombinant Adeno-Associated Virus Particle Titer Through Size Exclusion Chromatography with Multiangle Light Scattering and Collaborative Calibration of Standard Substances
Adeno-associated virus (AAV) is a promising gene therapy vector due to its high transduction efficiency, low pathogenicity, low immunogenicity, and the ability to mediate the long-term stable expression of exogenous genes. The viral particle titer is an essential quality attribute of recombinant ade...
Saved in:
| Main Authors: | , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2025-05-01
|
| Series: | Molecules |
| Subjects: | |
| Online Access: | https://www.mdpi.com/1420-3049/30/10/2170 |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1850256494148190208 |
|---|---|
| author | Dening Pei Xiang Li Hua Bi Wenhong Fan Heng Wang Manli Cui Xi Qin Chenggang Liang |
| author_facet | Dening Pei Xiang Li Hua Bi Wenhong Fan Heng Wang Manli Cui Xi Qin Chenggang Liang |
| author_sort | Dening Pei |
| collection | DOAJ |
| description | Adeno-associated virus (AAV) is a promising gene therapy vector due to its high transduction efficiency, low pathogenicity, low immunogenicity, and the ability to mediate the long-term stable expression of exogenous genes. The viral particle titer is an essential quality attribute of recombinant adeno-associated virus (rAAV) gene therapy products. Multiangle light scattering (MALS) is an important means of directly measuring the absolute molecular weight and distribution of macromolecular drugs. This study established and validated a method based on SEC-UV-MALS-RI tandem technology for accurately determining rAAV particle titers. The verification results indicated that the method exhibited good specificity, linearity, precision, accuracy, and durability. Several collaborative laboratories used this method to calibrate the standard substances needed for rAAV particle titer determination. The results suggested that combining the SEC-MALS method with standard substances enables the rapid and accurate measurement of the viral particle titers in rAAV gene therapy products. |
| format | Article |
| id | doaj-art-1866c82ccf58413caf2e7bd8eeac03d9 |
| institution | OA Journals |
| issn | 1420-3049 |
| language | English |
| publishDate | 2025-05-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Molecules |
| spelling | doaj-art-1866c82ccf58413caf2e7bd8eeac03d92025-08-20T01:56:38ZengMDPI AGMolecules1420-30492025-05-013010217010.3390/molecules30102170Methodological Study on Determination of Recombinant Adeno-Associated Virus Particle Titer Through Size Exclusion Chromatography with Multiangle Light Scattering and Collaborative Calibration of Standard SubstancesDening Pei0Xiang Li1Hua Bi2Wenhong Fan3Heng Wang4Manli Cui5Xi Qin6Chenggang Liang7National Institute for Food and Drug Control, No. 31 Huatuo St., Daxing District, Beijing 100050, ChinaNational Institute for Food and Drug Control, No. 31 Huatuo St., Daxing District, Beijing 100050, ChinaNational Institute for Food and Drug Control, No. 31 Huatuo St., Daxing District, Beijing 100050, ChinaNational Institute for Food and Drug Control, No. 31 Huatuo St., Daxing District, Beijing 100050, ChinaWaters Technology (Beijing) Co., Ltd., Building 1, No. 156, Jinghai 4th Road, Tongzhou District, Beijing 101111, ChinaWaters Technology (Beijing) Co., Ltd., Building 1, No. 156, Jinghai 4th Road, Tongzhou District, Beijing 101111, ChinaNational Institute for Food and Drug Control, No. 31 Huatuo St., Daxing District, Beijing 100050, ChinaNational Institute for Food and Drug Control, No. 31 Huatuo St., Daxing District, Beijing 100050, ChinaAdeno-associated virus (AAV) is a promising gene therapy vector due to its high transduction efficiency, low pathogenicity, low immunogenicity, and the ability to mediate the long-term stable expression of exogenous genes. The viral particle titer is an essential quality attribute of recombinant adeno-associated virus (rAAV) gene therapy products. Multiangle light scattering (MALS) is an important means of directly measuring the absolute molecular weight and distribution of macromolecular drugs. This study established and validated a method based on SEC-UV-MALS-RI tandem technology for accurately determining rAAV particle titers. The verification results indicated that the method exhibited good specificity, linearity, precision, accuracy, and durability. Several collaborative laboratories used this method to calibrate the standard substances needed for rAAV particle titer determination. The results suggested that combining the SEC-MALS method with standard substances enables the rapid and accurate measurement of the viral particle titers in rAAV gene therapy products.https://www.mdpi.com/1420-3049/30/10/2170adeno-associated virusviral particle titermultiangle light scatteringsize exclusion chromatography |
| spellingShingle | Dening Pei Xiang Li Hua Bi Wenhong Fan Heng Wang Manli Cui Xi Qin Chenggang Liang Methodological Study on Determination of Recombinant Adeno-Associated Virus Particle Titer Through Size Exclusion Chromatography with Multiangle Light Scattering and Collaborative Calibration of Standard Substances Molecules adeno-associated virus viral particle titer multiangle light scattering size exclusion chromatography |
| title | Methodological Study on Determination of Recombinant Adeno-Associated Virus Particle Titer Through Size Exclusion Chromatography with Multiangle Light Scattering and Collaborative Calibration of Standard Substances |
| title_full | Methodological Study on Determination of Recombinant Adeno-Associated Virus Particle Titer Through Size Exclusion Chromatography with Multiangle Light Scattering and Collaborative Calibration of Standard Substances |
| title_fullStr | Methodological Study on Determination of Recombinant Adeno-Associated Virus Particle Titer Through Size Exclusion Chromatography with Multiangle Light Scattering and Collaborative Calibration of Standard Substances |
| title_full_unstemmed | Methodological Study on Determination of Recombinant Adeno-Associated Virus Particle Titer Through Size Exclusion Chromatography with Multiangle Light Scattering and Collaborative Calibration of Standard Substances |
| title_short | Methodological Study on Determination of Recombinant Adeno-Associated Virus Particle Titer Through Size Exclusion Chromatography with Multiangle Light Scattering and Collaborative Calibration of Standard Substances |
| title_sort | methodological study on determination of recombinant adeno associated virus particle titer through size exclusion chromatography with multiangle light scattering and collaborative calibration of standard substances |
| topic | adeno-associated virus viral particle titer multiangle light scattering size exclusion chromatography |
| url | https://www.mdpi.com/1420-3049/30/10/2170 |
| work_keys_str_mv | AT deningpei methodologicalstudyondeterminationofrecombinantadenoassociatedvirusparticletiterthroughsizeexclusionchromatographywithmultianglelightscatteringandcollaborativecalibrationofstandardsubstances AT xiangli methodologicalstudyondeterminationofrecombinantadenoassociatedvirusparticletiterthroughsizeexclusionchromatographywithmultianglelightscatteringandcollaborativecalibrationofstandardsubstances AT huabi methodologicalstudyondeterminationofrecombinantadenoassociatedvirusparticletiterthroughsizeexclusionchromatographywithmultianglelightscatteringandcollaborativecalibrationofstandardsubstances AT wenhongfan methodologicalstudyondeterminationofrecombinantadenoassociatedvirusparticletiterthroughsizeexclusionchromatographywithmultianglelightscatteringandcollaborativecalibrationofstandardsubstances AT hengwang methodologicalstudyondeterminationofrecombinantadenoassociatedvirusparticletiterthroughsizeexclusionchromatographywithmultianglelightscatteringandcollaborativecalibrationofstandardsubstances AT manlicui methodologicalstudyondeterminationofrecombinantadenoassociatedvirusparticletiterthroughsizeexclusionchromatographywithmultianglelightscatteringandcollaborativecalibrationofstandardsubstances AT xiqin methodologicalstudyondeterminationofrecombinantadenoassociatedvirusparticletiterthroughsizeexclusionchromatographywithmultianglelightscatteringandcollaborativecalibrationofstandardsubstances AT chenggangliang methodologicalstudyondeterminationofrecombinantadenoassociatedvirusparticletiterthroughsizeexclusionchromatographywithmultianglelightscatteringandcollaborativecalibrationofstandardsubstances |