Pipeline Expression Platforms and the Future of Upstream Processing

In light of the fact that biopharma companies are experiencing increasing pressures to bring effective and safe products to market more quickly and at lower cost, constant innovation throughout the entire development and manufacturing process is inevitable. As a result, expression systems are also being scrutinized to help in the optimization of upstream processing efficiency.

To learn more about pipeline platforms and technologies, BioPharm International spoke with Michael A. Cunningham, associate director, Upstream Manufacturing Sciences and Technology, Life Sciences-Process Solutions at MilliporeSigma; Abhijeet Kohli and Natasha C. Lucki, PhD, both product managers at Thermo Fisher Scientific; Dr. Nicholas Holton, R&D manager at Leaf Expression Systems; Terence Ryan, chief scientific officer of iBio; Mark Emalfarb, chief executive officer of Dyadic International; and Dr. Fay Saunders, head of upstream mammalian cell culture, process development, at FUJIFILM Diosynth Biotechnologies, UK site.

BioPharm: What expression systems, that are in the pipeline or being developed, do you foresee as having a significant impact on upstream processing?

Cunningham (MilliporeSigma): The CHOZN expression platform is attractive given that it is part of a full upstream platform complete with a vector, media, feeds, and protein quality attribute-modifying supplements. Next-generation CHOZN expression platforms are in development, which should shorten cell line development time, and rapidly provide materials (i.e., cell clones and/or recombinant product) for early characterization studies. Additionally, some current CHOZN expression platform development activities are focusing on optimizing the controlled and reproducible targeted insertion of the candidate transgene.

Kohli and Lucki (Thermo Fisher Scientific):Thermo Fisher Scientific has recently expanded the suite of Gibco ExpiCHO solutions to support late stage/stable production workflows. The Gibco ExpiCHO Stable Production Medium supports the growth of ExpiCHO-S stable clones and facilitates scale-up as drug candidates move to late development and beyond. Crucially, this production medium harmonizes with the other components of the Gibco ExpiCHO platform and therefore enables an ideal transition from early-to-late-stage workflows without having to deal with the unpredictability issues commonly encountered when switching cell lines, which will help to improve quality and support faster development timelines.

There is also a need to streamline the development process a bit more, and there are a few ways to do this. Instead of undertaking the random integration of genes into cell lines, it’s possible to go after site-directed gene integration. This approach involves identifying a locus in the genome of the cell that can be stably integrated. Using this method, it’s possible to reduce the amount of post-integration selection work that is required, ultimately accelerating timelines.

Holton (Leaf Expression Systems): Despite having received far fewer R&D resources over the past few decades, plant systems have still managed to become competitive with conventional systems, and indeed surpass them in many areas. This is a testament to their distinctive advantages. As funding for further optimization of plant systems grows over the coming years, and as an increasing number of plant-produced pharmaceuticals are granted approval, we expect further gains in the efficiency of plant systems to be made and their share of the market to rapidly increase.

Ryan (iBio):There is an industry-wide desire (as well as need) to lower costs and speed of product development, and each of the expression technologies is being exploited to meet these demands. Lower costs are being driven by payor concerns as well as the need to open other markets worldwide, and speed is driven by the needs of patients as well as shortening time-to-market to recoup costs and maintain a patent window.

The nature of the improvements in each expression system is unknown until disclosed, but at iBio, we similarly work to expand our portfolio possibilities, improve the speed and efficiency of our technology, improve our ability to manufacture biologics which fail in other systems, and make our technology affordable and accessible around the globe, thereby improving patient access to needed medicines.

Emalfarb (Dyadic International): Dyadic (together with a growing number of pharmaceutical and biotech companies) believe that with the advent of and rapid pace of scientific advances in synthetic biology and the associated genetic tools to engineer cell lines that industrially proven hyper productive cell lines like Dyadic’s C1 cell line, which is based on the fungus Myceliophthora thermophila can be a potentially significant biopharmaceutical gene expression platform. 

The C1 microorganism, which enables the development and large-scale manufacture of low-cost proteins, has the potential to be further developed into a safe and efficient expression system that may help speed up the development, lower production costs, and improve the performance of biologic vaccines and drugs at flexible commercial scales.

Saunders (FUJIFILM):Cell free synthesis, despite being a well-established technique in research and applied biology, has been emerging for a few years in the field of protein production, offering impressive yields and low time and cost requirements. Another potential avenue for companies to improve efficiencies may lay in the use of site-specific integration to enable a more predictable expression and, as such, require fewer screening efforts. New host cell lines are coming to the fore that can positively impact the development process.

BioPharm: Are there any changes to regulations or approvals of expression technology/processes that could benefit and/or hinder the industry?

Emalfarb (Dyadic International): Well one thing for sure is that if the government, in conjunction with FDA, would put in place laws and procedures to fast track biologics that are produced with alternative cell lines, there is the potential for change in the cost structure of biomanufacturing. When you think about the impact a reduction in biomanufacturing costs could mean to making biologics more accessible and affordable to the millions of patients suffering or dying from diseases globally, there really is no excuse not to provide incentives to the pharma and biotech industry to speed up the adoption and use of alternative cell lines that grow faster, need much lower cost media, and produce biologics at higher levels.

Saunders (FUJIFILM):There has been a reduction in the barriers to market entry, and there are clear pathways that can be followed for the products that previously would have had to go through individual assessments; biosimilars are a good example. However, technology and the molecules being developed are continually evolving so regulations will need to keep up with this.  

Cunningham (MilliporeSigma): As optimization of protein expression continues to improve, particularly with the emergence of treatments for niche patient populations, characterization of pharmacogenomics is likely to become a greater regulatory focus to ensure product efficacy and safety.

Holton (Leaf Expression Systems): A number of plant-derived products have already been approved or are currently in clinical trials, take Medicargo’s Phase III quadrivalent influenza vaccine (QIV) for example. 

Through industry engagement, regulation qualifying guidelines are constantly changing and being rewritten to take in to account new expression systems and advanced therapies. Ultimately the downstream processing is the same as other ‘traditional’ expression technologies, which significantly reduce the risks of product non-compliance from a regulatory perspective. 

Kohli and Lucki (Thermo Fisher Scientific):Regulations are evolving to adjust to the next generation of biologics, such as multi-specifics, Fc-fusions, and viral vectors, entering the pipeline. These new molecules often have different biophysical and biochemical properties than the traditional mAbs [monoclonal antibodies] that have been the main macromolecule in the pipeline for many years. There seems to be an open line of communication between the developers and the regulators to establish new processes and assays tailored to these new categories of biologics to ensure compliance, and, ultimately grant approval.

Ryan (iBio):We have found that FDA and other international regulatory bodies are agnostic about expression systems as long as the manufactured proteins are safe, effective, and reproducible. The use of alternative expression systems will likely benefit from more regularized definitions of what ‘biosimlar’ and ‘biobetter’ drugs are, and the analytical methods required to characterize and qualify these increasingly important categorizations.