On-demand manufacturing of clinical-quality biopharmaceuticals

Abstract

Conventional manufacturing of protein biopharmaceuticals in centralized, large-scale single-product facilities is not well-suited to the agile production of drugs for small patient populations or individuals. Solutions for small-scale manufacturing are potentially more nimble, though previous systems are limited in both process reproducibility and product quality, owing to complicated means of protein expression and purification 1 – 4 . We describe an automated bench-top multi-product manufacturing system, called Integrated Scalable Cyto-Technology (InSCyT), for the end-to-end production of hundreds to thousands of doses of clinical-quality protein biologics in about three days. We also demonstrate that InSCyT can accelerate process development from sequence to purified drug in 12 weeks. We produced hGH, IFNα-2b, and G-CSF using highly similar processes on InSCyT and found that the purity and potency of these products is comparable to that of marketedreference products.

Comparative genome‐scale analysis of Pichia pastoris variants informs selection of an optimal base strain

“…Protein concentrations of rhGH and rhG‐CSF were determined using sandwich enzyme‐linked immunosorbent assay (ELISA) as described elsewhere (Crowell et al, ). rhGH‐specific antibodies were used at concentrations of 5 µg/ml capture (ab1954; Abcam, Cambridge, UK) and 0.6 µg/ml secondary/detection (ab1956; Abcam).…”

“…Quantitative polymerase chain reaction (PCR) was performed according to PowerUp SYBR Green kit instructions (Thermo Fisher Scientific) using a Roche LightCycler 480. β-Actin, rhGH, rhG-CSF, and trastuzumab ORFs were PCR amplified and serially diluted to generate a standard curve (see Table S2 for primers). Experimental design and absolute copy number quantification were performed according to best practices (Abad et al, 2010) Protein concentrations of rhGH and rhG-CSF were determined using sandwich enzyme-linked immunosorbent assay (ELISA) as described elsewhere (Crowell et al, 2018). rhGH-specific antibodies were used at concentrations of 5 µg/ml capture (ab1954; Abcam, Cambridge, UK) and 0.6 µg/ml secondary/detection (ab1956; Abcam).…”

On-demand, cell-free biomanufacturing of conjugate vaccines at the point-of-care

“…Importantly, by enabling portable production of bioconjugate vaccines, iVAX addresses a key gap in both cell-free and decentralized biomanufacturing technologies. Production of glycosylated products has not yet been demonstrated in cell-based decentralized biomanufacturing platforms (Crowell et al, 2018;Perez-Pinera et al, 2016) and existing cell-free platforms using E. coli lysates lack the ability to synthesize glycoproteins (Boles et al, 2017;Murphy et al, 2019;Pardee et al, 2016;Salehi et al, 2016). While glycosylated human erythropoietin has been produced in a cell-free biomanufacturing platform based on freeze-dried Chinese hamster ovary cell lysates (Adiga et al, 2018), this and the vast majority of other eukaryotic cell-free and cell-based systems rely on endogenous protein glycosylation machinery.…”

“…In addition, the ability to rapidly produce vaccine candidates in vitro provides a unique means for rapidly responding to pathogen outbreaks and emergent threats. As a result, we believe that the iVAX platform, along with an emerging set of technologies with the ability to synthesize biomedicines on-demand (Adiga et al, 2018;Boles et al, 2017;Crowell et al, 2018;Murphy et al, 2019;Pardee et al, 2016;Perez-Pinera et al, 2016;Salehi et al, 2016), has the potential to promote increased access to complex, life-saving drugs through decentralized production. All other authors declare no competing financial interests.…”

The new strategies to overcome challenges in protein production in bacteria

“…From 2011 to 2016, 62 new biologics were approved by the FDA (Lagass e et al, 2017). Today, this production is centralized and large-scale, but in the future, small-scale manufacturing adapted to individual needs of smaller patient populations may become the standard (Crowell et al, 2018). An aim of the biological revolution will be to produce functional protein in a costefficient manner.…”