A review.The current developments in the purification of four of the most popular and emerging gene therapy vectors, i.e., adenoviral vectors, adeno-associated viral vectors, lentiviral vectors, and plasmid DNA, are described.Recent developments in the application of membrane ion exchange chromatog. technol. for the purification of such vectors are discussed.

Membrane chromatography purification of adenovirus and adeno-associated viral vectors for gene therapy

Author: Lajmi, Ajay

Developing scaleable processes for the purification of recombinant adenoviral (rAd) and adeno-associated viral (rAAV) vectors is essential as gene therapy advances from the laboratory to clin. trials.It is critical that contaminants such as RNA, chromosomal DNA, and host cell proteins are efficiently removed from vector preparations that are used for clin. trials.Suitable purification of substantial quantities of rAd and rAAV vectors by conventional methods is limited, especially when d.-gradient centrifugation is involved.Membrane chromatog. offers an alternative and efficient method for the capture of viral vectors in a disposable format with throughputs of 10 to 100-fold higher than beaded chromatog. media.A simple scaleable process was developed for the purification of rAd and rAAV vectors for gene therapy.The process utilizes high dynamic binding capacity of strong anion exchange membranes for chromatog. capture of intact, infectious viral particles.A total of 5 x 1013 rAd viral particles could be efficiently captured and eluted in 70% yield from a strong anion exchange membrane chromatog. unit.Analyses of the purified products that will be presented indicate that membrane chromatog. provides high throughput and scalable purification method for producing recombinant viral vectors suitable for clin. trials.

Pharmaceutical Technology

Scalable membrane ion-exchange chromatography purification of an antisense oligonucleotide

Author: Lajmi, Ajay R.

A 20-mer phosphorothioate oligonucleotide was efficiently captured and purified on a 10-mL bed volume strong anion-exchange membrane unit.The purification cycle was completed in less than 30 min with >99.0% purity anal overall yields of 83%.

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