The future
of healthcare
is bio.
Cell-Free fields of application
“Point of care” (POC) biomanufacturing consists on the development of portable, fully-integrated and closed platforms that incorporate biological materials like yeast cells, bacteria or cell lysates, which are used for the bioproduction of specific products like therapeutic proteins, interferons, recombinant hormones and vaccines at the point of care location.
The advantage of this point of care approach to medicine and biomanufacturing is that biologics can be produced in a short time and with lightweight devices, allow patients to be treated with biologics in remote locations, on battlefields, in emergency situations, or in underdeveloped areas with poor infrastructure.
How it works?
The basic components of “point-of-care” bioproduction devices are:
. a biological expression system designed to express the biotherapeutic product in response to programmable signals,
. an integrated microfluidic platform capable of controlling the biomanufacturing process quickly and flexibly, enabling operations to take place remotely.
The biomanufacturing system can be designed to produce more than one biological product simultaneously, provided that they are compatible with one another, and purification methods are integrated into the system. Furthermore, if cell-free extracts are used, they can be stored in freeze-dried form and rehydrated with water, prior to being incubated using body heat to activate the extractable components, demonstrating the remarkable portability of such systems.
Shifting to point of care biomanufacturing
Centralized biomanufacturing in highly specialized facilities has been the dominating model for the production of bioproducts. However, this model imposes limitations in accessibility and scalability, representing a bottleneck that struggles to keep up with the increasing demand for biotherapies. Recently, COVID-19 pandemic evidenced the need for agile biomanufacturing and distribution frameworks to deliver biologics and biopharmaceuticals worldwide.
POC biomanufacturing, enabled by recent developments in technology, can help to overcome these limitations. One illustrative example is the production of autologous gene and cell therapies, where patient cells have to be transported to the manufacturing facility, processed and returned to the patient, in a process that may take weeks. By shifting to POC bioproduction, the time needed to make the treatment available to the patient and the complexity in logistics are reduced, increasing the accessibility and affordability of treatments [1].
Recently, a microfluidic chip able to produce clinical doses of CAR-T cells for cancer therapy was developed [2]. The ultra-small (with the size of a pack of cards), automated and closed system represents the first use of a microbioreactor for autologous cell therapy, and enables the production of CAR-T cells with the same efficacy of traditional methods at a lower cost. The developed microfluidic chip allowed to diminish the production time in 30-40%, and the small starting cell numbers required, compared to existing larger automated manufacturing platforms, enabled to reduce the amounts of isolation beads, activation reagents and vectors needed per production run, as well as the cell culture volume (2 ml). Besides the reduction in reagent cost, this proposal is also beneficial to pediatric patients who have low or insufficient T-cell numbers to produce therapeutic doses of CAR-T cells.
Challenges
The change towards POC bioproduction is considered critical and an emergence by the European Medicines Agency (EMA) and Food and Drug Administration (FDA) [3,4], although it is assumed that it will be slow. The existing traditional biomanufacturing has been frequently sufficient but, gradually, the need to replace existing production lines and to proceed to local manufacturing will open opportunities to change to POC production. Another challenge is to assure consistent quality in products biomanufactured by POC systems.
Prefabricated portable cleanrooms and automated processes will allow the standardization of equipments and process, and digital technology will enable to connect the data and quality systems of different locations [5]..
Additionally, scaling-down the upstream processing and downstream unit operations performed in traditional bioprocesses, such as purification of the synthesized bioproduct, may arise some difficulties in POC biomanufacturing [6].
Cell-free systems and point of care bioproduction
To address the challenge of downscaling the upstream processing in POC bioproduction, cell-free systems can efficiently replace the traditional cell culture process. In cell-free synthesis, a lyophilized solution containing cellular components is used in the bioreactor, together with a plasmid containing the sequence for the target protein [6].
A successful example of using cell-free systems in POC bioproduction is the Bio-MOD system, that employs a cell-free method for end-to-end continuous manufacturing of biological drug substances [7]. The portable (suitcase-sized) cell-free bioprocessing system integrates machine learning to produce proteins with increased and consistent purity and quality wherein such proteins are prepared on-demand. The system can be scaled-out to enable working with larger volumes.
The machine learning algorithm extracts sufficient data from the process sensors and analytical measurements to ensure the consistency of the process. The algorithm is also able to detect any deviation from the expected, which increases the safety level, compared to the current method of testing a product after it is made.
Therapeutics-on-a-chip system was also recently developed and consists on a microfluidic chip containing a cell-free expression system, which was used to produce and purify green fluorescent protein and cecropin B with a high purity and at therapeutically relevant concentrations at a low cost (Figure 1) [8]. The authors used fresh and lyophilized materials for cell-free synthesis of the proteins and applied immunoprecipitation for highly efficient, on-chip protein purification.
Figure 1 – The integrated system for cell-free protein synthesis and purification.
Publications
References
To know more
Nature communications
Synthetic biology and microbioreactor platforms for programmable production of biologics at the point-of-care
Sciences Advances
On-demand biomanufacturing of protective conjugate vaccines
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Cell-Free Systems Applications