Md Nasimuzzaman

Accomplishment  Successfully owned and executed multiple client projects.  Recruited and managed a team of scientists, overseeing process design and optimization using DOE studies.  Led development of platforms for scalable LV and AAV manufacturing and analytics for gene and cell therapy drugs in compliance with the cGMP regulations.  Effectively managed collaborative projects, vendors, CROs, and CDMOs.  Led a cross-functional team in deviation investigation and root cause analysis, and CAPA. AAV Vector Process Development and Manufacturing  Optimized cell culture and transfection for AAV vector in HEK293 suspension cell culture.  Optimized parameters for AAV purification, including affinity chromatography, AEX for empty-full separation, tangential flow filtration (TFF), and formulation.  Implemented Tech transfer the process to GMP facility for manufacturing clinical-grade vectors. LV Vector Process Development and Manufacturing  Optimized upstream processes for LV vector in HEK293 in shake flasks and bioreactors.  Optimized downstream parameters for LV purification, utilizing AKTA chromatography, tangential flow filtration (TFF), and formulation. Analytical Method Development and Characterization of Viral Vectors  Designed and executed experiments to evaluate titers (genomic, infectious, particle), purity, and stability of LV vectors using qPCR, ddPCR, ELISA, and flow cytometry.  Assessed transgene expression for gene and cell therapy products.  Analyzed experimental data, interpreted results, and generated scientific reports to support manufacturing and regulatory requirements. Cell Therapy & Potency Assay • Led development of autologous CAR-T therapy (CD19) including T-cell activation, transduction, expansion, and functional characterization. • Designed and implemented cell-based potency assays, including tumor killing and cytokine release models. • Advanced understanding of T cell phenotype, function, and engineering strategies.

Accomplishments  Spearheaded gene therapy preclinical research, driving innovation and scientific advancements.  Successfully established an independent research program and laboratory from the ground up, ensuring state-of-the-art facilities and resources.  Performed a risk assessment of CQAs, CPPs, and CMAs for AAV drug products, ensuring alignment with QTPP as part of the CMC of drug products.  Recruited, trained, and effectively managed a team of scientists, fostering collaborative and high-performing environment.  Diligently managed budgets and project timelines across multiple initiatives.  Managed external collaboration with CROs and CDMOs. AAV Vector Production and Purification for Gene Therapy Applications  Optimized AAV Production - Leveraged Sf9 and Baculovirus alongside HEK293-plasmids systems to optimize AAV production. - Fine-tuned critical process parameters for upstream processes in shake flasks and bioreactors.  Streamlined Downstream Processes and Analytics of AAV - Optimized parameters for downstream processes, employing affinity chromatography (AKTA system) and TFF. - Developed and validated analytical methods for characterizing AAV vectors and assessing transgene expression. - Designed and executed experiments to evaluate the potency and titers, purity, and stability of AAV using qPCR, ELISA, CE-SDS, HPLC, and flow cytometry. Developed novel mouse line using CRISPR-Cas9 mediated genome editing  Designed guide RNA and donor oligo sequences containing LoxP sites.  Led generating novel PAR-4fl/fl mouse in collaboration with the transgenic animal core facility. Generated PAR-1 knockout (KO) cell line using CRISPR-Cas9 mediated genome editing  Cloned guide RNA and Cas9 sequences into lentivirus (LV) vector plasmid.  Produced LV vector in HEK293T cells by PEI-mediated transfection.  Infected the LV vector into HT-29 cell to generate PAR-1 KO single-cell clones.

Accomplishments  Served as an SME for upstream, downstream, and analytics of AAV, Adv, LV, and RV.  Developed scalable processes in compliance with the cGMP regulations.  Manufactured viral vectors for internal and external clients.  Mentored junior scientists and troubleshot their experimental challenges.  Collaborated as a trusted CRO and CDMO partner with leading GCT biotech companies. Process Development and manufacturing of AAV Vector with HEK293 Cells  Optimized upstream processes for AAV vector production via PEI-mediated transfection of HEK293 cells.  Enhanced downstream processes for AAV purification using chromatography and TFF.  Performed process characterization, and stability studies in support of establishing process understanding and control strategies.  Characterized AAV through SDS-PAGE-silver staining, immunoblotting, titrated with qPCR, and analyzed gene expression in transduced cells as part of the potency evaluation. Process Development of AAV Vector Using Insect Cell Culture  Developed AAV production utilizing Sf9 and Baculovirus (Bac-to-bac) system.  Generated Baculovirus-infected insect cells (BIIC) for AAV production.  Advanced upstream processes for AAV in shake flasks and bioreactors.  Enhanced downstream processes for AAV purification using affinity chromatography.

Gene Therapy for Beta-Thalassemia & Sickle Cell Anemia in Mouse Models  Conducted preclinical safety and efficacy studies to characterize LV vectors and genemodified cells for IND-enabling studies.  Led optimization of vector design and transgene expression and evaluated vector potency in animal models.  Alleviated beta-thalassemia in mouse models using LV-globin vector transduction in hematopoietic stem cells and bone marrow transplantation.  Analyzed viral vector biodistribution in cells using next-generation sequencing (NGS).  Developed robust potency assays, assessed risks, and recommended advanced technologies to enhance gene therapy research. Genotoxicity Analysis of LV Vector with AAV-Mediated Genome Editing  Designed, produced, and purified a novel AAV-HMG-GFP vector.  Generated recombinant cell clones via AAV vector transduction, homologous recombination, and Cre-loxP-mediated cassette exchange.