Exploiting heparin-binding properties of MoMLV-based retroviral vectors for affinity chromatography.
Chromatography is deemed the most promising technology for large-scale purification of viral vectors. The authors have previously shown that heparin affinity chromatography could be successfully employed for the purification of VSV-G pseudotyped Moloney murine leukemia virus (MoMLV)-derived vectors giving excellent results in terms of recovery of active particles, reproducibility and selectivity. In this study, the authors examined whether the ability of retrovirus particles to specifically bind to heparin ligands is restricted to VSV-G pseudotypes produced by 293-based packaging cells. It is shown that VSV-G deficient retrovirus particles are captured by a heparin chromatography column as efficiently as VSV-G containing particles. Most strikingly, RD114 pseudotyped retrovirus particles derived from a HT1080-based cell line were found to bind heparin with the same affinity as 293-derived VSV-G pseudotypes. RD114 pseudotyped retrovirus particles were successfully isolated using heparin affinity chromatography obtaining good recoveries of functional particles (43%). These results indicate that heparin affinity chromatography can be extended to the purification of retroviral vectors produced by different packaging cell lines independently of the Env-protein used for pseudotyping.
Segura
María de las Mercedes
MM
Department of Chemical Engineering and Centre de Recherche sur la Fonction, la Structure et l'Ingénierie des Protéines, Université Laval, Que., Canada G1K 7P4.
Kamen
Amine
A
Lavoie
Marie-Claude
MC
Garnier
Alain
A
D002460:Cell Line; D002846:Chromatography, Affinity / Q000379:methods*; D005822:Genetic Vectors; D006493:Heparin / Q000378:metabolism*; D006801:Humans; D008856:Microscopy, Fluorescence; D008979:Moloney murine leukemia virus / Q000235:genetics*; D014161:Transduction, Genetic
D006493:Heparin