Our research focuses on system engineering in life sciences. It covers biomanufacture innovations in gene and cell therapy; protein therapeutics; and drug formulation and delivery. We are aiming at developing innovative systems technology that can improve drug development e ciency and manufacturing productivity, and developing transformative diagnostic systems and tools for selected diseases with chemometrics framework. Integration of medical devices with multivariate statistical method is also being explored to develop practical diagnostic tools
Exploring alternative hosts for Adeno-associated virus production
Adeno-associated virus (AAV)-based vector is among the most prominent delivery methods for gene therapy
treatments. Depending on the producer, the AAV products can contain contaminating transforming factor from
the producer cell lines, including the adenovirus E1 gene in HEK293 cells and human papillomavirus E6 and E7
oncogenes in HeLa cells. These oncogenes can integrate into the host cell genome and induce tumors. To mitigate
this risk, we explore cell lines immortalized by the non-oncogenic human Telomerase Reverse Transcriptase
(hTERT) gene as alternative AAV producers. The expression of hTERT maintains the size of the telomeres. As a
result, in cells where shortening of the telomeres is the only obstacle to immorality, hTERT alone can immortalize
the cells without inducing cancer-associated changes. Six hTERT cell lines have been selected: hTERT RPE1, BJ-5ta,
hTERT Lung Fibroblast, hTERT HME1, RPTEC/TERT1 OTC2, and UMB1949. Among the six cell lines of interest, HME1
with jetOPTIMUS was identi ed as the most promising AAV host. The cell line yielded 6E10 GFP copies and 4E9
capsids per litter. However, for HME1 to be a successful oncogene-free AAV producer, its encapsulated genome
and capsid yield needs to be improved to be comparable to industry standard. Preliminary data shows that the
transcription level of Rep and Cap gene in HME1 is much lower than that in control cells (293T), thus redesigning
the Rep/Cap plasmid with a stronger promoter is a possible solution. Furthermore, a redesign of the E1A/B
plasmid is necessary to retain only the essential regions for rAAV production while eliminating its oncogenic
e ects.
Gene therapy media optimization
The production of recombinant adeno-associated virus (rAAV) for gene therapy applications is experiencing rapid expansion. This surge is driven by the increasing demand for rAAV-based gene therapy products, which necessitates advancements in the manufacturing processes and technologies to achieve scalable and high-yield production of rAAV vectors for therapeutic gene delivery. This research topic mainly focuses on understanding the main role of media components and developing improved media containing selected media components for maximized rAAV yield and improved quality attributes.
CRISPR-CAS9 mediated genome engineering of HEK293 cells for improved RAAV manufacturing yield
The development of gene therapies based on recombinant adeno-associated viruses (rAAVs) has grown exponentially, so the current rAAV manufacturing platform needs to be more e cient to satisfy rising demands. Viral production exerts great demand on cellular substrates, energy, and machinery; therefore, viral production relies heavily on the physiology of the host cell. Transcriptomics, as a mechanism-driven tool, was applied to identify signi cantly regulated pathways and to study cellular features of the host cell for supporting rAAV production. This research topic mainly focuses on utilizing transcriptomics to reveal molecular signatures for rAAV production and modulating the yield by medium supplementation and cell line engineering strategies hypothesized based on the transcriptomic data.
Develop an inducible stable packaging/ producer cell line for RAAV production via site specific integration
rAAV-mediated gene therapy manufacturing is a quickly growing segment of the pharmaceutical market. Developing innovative stable cell lines will greatly enhance the productivity and product quality attributes for rAAV production, making it more widely accessible to people.
Fluorescence-based multi-step IEX-HPLC method for quantification of full, partial and empty capsid in AAV products
The research focuses on analytical method development for rAAV capsid di erentiation including full, partial, and empty capsids. Capsid impurities including empty and partial capsids during rAAV manufacturing pose great hurdles in gene therapy in terms of product quality, e cacy, and safety; therefore, industry and the regulatory need a tool providing more accurate and comprehensive capsid pro les in the nal gene therapy products. This applies to the same for the downstream process where full capsids with therapeutic e ects are separated from the empty and partially lled capsids.
AI-enabled Hyperspectral Imaging for the Real-Time Monitoring of Cell Culture Metabolites
In terms of production methods we are progressing from production in adherent cells in Cell factories and Hyperstacks which require a lot of space and scale out to suspension cell platforms like batch, fed-batch and perfusion using transient transfection followed by the use of stable suspension cell lines in perfusion mode in order to obtain ever higher yields. Increasing Process Analytical Technologies such as the use of Near Infrared spectroscopy to better understand the process and finally the use of machine learning and AI to increase automation of the process.