Biotechnology start-ups are often advised to think early about process design to avoid hiccups during scaleup. A clinical-stage gene-therapy developer has taken this approach to heart, saying they’ve designed their processes using Quality-by-Design.

Susan D’Costa, PhD, executive vice president and global head of technology at Alcyone Therapeutics, which has two candidate therapies in or approaching clinical development, says the company is investing early in developing a flexible upstream manufacturing process.

“Companies often say they’d rather use their early development frugally on CMC [Chemistry Manufacturing and Controls] and get into the clinic with a ‘bench-scale process,’ show proof of efficacy and dollar safety, and then build big,” says D’Costa.

But, she adds, this approach risks process impurities hampering their later development by affecting patient safety, drug tolerance, and effectiveness.

“Process and product impurities, leading to cost and safety concerns, are some of the biggest headwinds in the industry right now, and are a challenge to gene therapy investors who don’t know whether a process is safe to go forwards,” she explains.

D’Costa, who is speaking at the 14th Annual Bioprocessing Summit in Boston this week, explains that Alcyone are working to overcome these problems by designing robust processes early.

Transient transfection of mammalian cells

D’Costa explains that the company assessed the benefits of several techniques for manufacturing AAV for their gene therapy, including using baculovirus in insect cells, and a packaged cell line where the information required to produce AAV is pre-written into the chromosomes of the cells. Having considered the likely difficulties of each approach, D’Costa and her team opted for transient transfection of mammalian cells using two or three plasmids.

“We’ve decided to use transient transfection as our process because that’s the industry technique we think right now gives us the best bang-for-our-buck,” she notes.

The company has used a similar design process to develop a potency assay for testing their treatment for Rett syndrome, a rare genetic neurological and development disorder. According to D’Costa, the assay monitors for the expression of a short non-coding RNA, the mechanism of action used in Alcyone’s X-reactivation gene therapy platform, rather than the expression of the MeCP2 protein, the gene mutated in Rett syndrome.

This approach means the same assay can be used for monitoring the upregulation of mutated genes in other X-chromosome-linked diseases and for testing future products in their pipeline.

“If we looked at the MeCP2 expression, that would just focus on Rett syndrome,” she tells GEN. “By monitoring the expression of that non-coding RNA, we don’t need to reinvent [anything].”

D’Costa claims their approach leads to faster and more efficient commercial development. “As soon as we have a safety and efficacy signal, we can scale up, make the product, and move straight into pivotal registration.”