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Klyushnichenko V, Bernier A, Kamen A, et al. Improved high-performance liquid chromatographic method in the analysis of adenovirus particles. J Chromatogr B Biomed Sci Appl. 2001;755(1):27-36doi: 10.1016/s0378-4347(00)00597-1.
Klyushnichenko, V., Bernier, A., Kamen, A., & Harmsen, E. (2001). Improved high-performance liquid chromatographic method in the analysis of adenovirus particles. Journal of chromatography. B, Biomedical sciences and applications, 755(1), 27-36. https://doi.org/10.1016/s0378-4347(00)00597-1
Klyushnichenko, V, et al. "Improved high-performance liquid chromatographic method in the analysis of adenovirus particles." Journal of chromatography. B, Biomedical sciences and applications vol. 755,1 (2001): 27-36. doi: https://doi.org/10.1016/s0378-4347(00)00597-1
Klyushnichenko V, Bernier A, Kamen A, Harmsen E. Improved high-performance liquid chromatographic method in the analysis of adenovirus particles. J Chromatogr B Biomed Sci Appl. 2001 May 05;755(1):27-36. doi: 10.1016/s0378-4347(00)00597-1. PMID: 11393714.
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J Chromatogr B Biomed Sci Appl. 2001 May 05;755(1):27-36. doi: 10.1016/s0378-4347(00)00597-1.

Improved high-performance liquid chromatographic method in the analysis of adenovirus particles.

Journal of chromatography. B, Biomedical sciences and applications

V Klyushnichenko, A Bernier, A Kamen, E Harmsen

Affiliations

  1. Biotechnology Research Institute, NRC, Montreal, Quebec, Canada. [email protected]

PMID: 11393714 DOI: 10.1016/s0378-4347(00)00597-1

Abstract

We developed a HPLC method on a novel continuous bed matrix (UNO Q, Bio-Rad) for the direct quantification of adenoviral type 5 (Ad5) particles produced in 293S Human Embryonic Kidney cells and compared this with an existing HPLC method on a conventional ion-exchange resin (Resource Q, Pharmacia). The 293S cell extract contained large amounts of DNA. This contaminated the viral peak on the Resource Q column and only after Benzonase treatment was it possible to quantify the viral particles in the cell extract. In contrast, the virus peak on the UNO Q column was resolved from the DNA which eliminates the need for pretreatment of the sample with Benzonase. Cross-analysis of the Ad5 fraction from the UNO Q column using a size-exclusion HPLC column revealed no additional contaminating peaks. We conclude that the purity of the Ad5 virus peak on the continuous bed matrix UNO Q column was superior to the purity of the virus on the conventional Resource Q column, which is essential for reliable quantification.

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