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

被引：42

作者：

Klyushnichenko, V ^{[1
]}

Bernier, A ^{[1
]}

Kamen, A ^{[1
]}

Harmsen, E ^{[1
]}

机构：

[1] Natl Res Council Canada, Biotechnol Res Inst, Bioproc Sector, Anim Cell Technol & Downstream Proc Grp, Montreal, PQ H4P 2R2, Canada

来源：

JOURNAL OF CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES | 2001年 / 755卷 / 1-2期

基金：

加拿大自然科学与工程研究理事会;

关键词：

adenovirus particles; high-performance liquid chromatography analysis;

D O I：

10.1016/S0378-4347(00)00597-1

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

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. Published by Elsevier Science B.V.

引用

页码：27 / 36

页数：10

共 23 条

[1] Green fluorescent protein as a real time quantitative reporter of heterologous protein production [J].