Pharmaceutical Technology - May 2020

Pharmaceutical Technology – Biologics and Sterile Drug Manufacturing

Issue link: https://www.e-digitaleditions.com/i/1248960

Contents of this Issue

Navigation

Page 23 of 42

24 Pharmaceutical Technology BIOLOGICS AND STERILE DRUG MANUFACTURING 2020 P h a r mTe c h . c o m Manufacturing V iral gene therapy has gained momentum after recent ap- provals and commercial successes with novel therapies based on chimeric antigen receptor T cell (CAR-T) or di- rect virus-mediated gene modification. Viral vectors such as adeno-associated viruses (AAV) and lentiviruses are the foundation of such therapies, and their manufacture is not without challenges. With the growing demand for these vectors and the desire to target large organs, higher virus titers are required; higher titers enable the number of batches to be minimized and reduces the cost of manufacturing. Lentiviruses are characterized by their diploid, single-stranded, positive sense RNA genomes, which are transcribed into viral DNA in the host cell and integrated into the genome. Because of this prop- erty, these viruses can be modified for use as replication incompetent vectors to deliver genes of interest to mammalian cells. Lentiviruses can also be used to package larger transgenes compared to other vectors, such as AAV. While lentiviral vector production can vary among developers and manufacturers, it follows a typical process. Once transiently transfected with four plasmids containing lentivirus structural pro- tein genes and the gene of interest (GOI), human embryonic kidney (HEK)-293 or HEK-293T cells are cultivated to produce viruses to a desired titer. The harvest bulk is then passed through a series of fil- tration and chromatography steps (Figure 1). Clarification is typically the first step of the downstream process and is a critical unit opera- tion that impacts subsequent steps and, ultimately, virus recovery. João Marques is senior scientist, and Paul Carter is scientific leader, Downstream Vector; both are at Downstream Process Development at the Cell & Gene Therapy Platform CMC, Medicinal Science and Technology, R&D at GlaxoSmithKline. Matthew Dillingham is a senior process development scientist, Paul Beckett is senior technology manager, Purification Technologies, Adina Paun is account manager pharma processing, Youness Cherradi is senior process development scientist, and Anissa Boumlic * , anissa. boumlic@milliporesigma.com, is associate director Vaccines and Viral Therapies Segment, all at MilliporeSigma. * To whom all correspondence should be addressed. BLUEBAY2014 - STOCK.ADOBE.COM Optimizing Viral Vector Manufacturing for Gene Therapy João Marques, Matthew Dillingham, Paul Beckett, Adina Paun, Youness Cherradi, Paul Carter, and Anissa Boumlic Synthetic depth filters are a suitable option to provide a high-performance and scalable single-step clarification process.

Articles in this issue

Links on this page

Archives of this issue

view archives of Pharmaceutical Technology - May 2020 - Pharmaceutical Technology – Biologics and Sterile Drug Manufacturing