Brijesh Garg

Managed exon skipping biomarker assays at CROs to timely deliver Phase 1 clinical data; transferred/qualified ddPCR and RT-PCR assays to support Phase 1 trial. Supervising Anti-Drug Antibody (ADA/immunogenicity, ELISA) MD/MV work at a CRO to detect antibodies in NHP/clinical samples; handling antibody generation externally to procure positive control/s. Managing/developing/validating PD biomarker WB, IHC, PCR assays externally to support Phase 2 study; developed ddPCR, JESS/WB internally to quantify PD marker in preclinical samples; supporting pre-IND/IND letter writing/submission. Developed qPCR/RT-PCR assays internally/externally to quantify gene transcripts for a Neuromuscular disorder; screened/finalized/managing a CRO for splicing multiplexing Next-Generation Sequencing assay to support a Phase 1 clinical trial. Established biology/bioanalytical lab at PepGen; searched/finalized CROs/vendors for critical assays/reagents; working with Operations/Finance/Legal teams to set-up MSA/CDA/SOW/Coupa; helping with the interviews; finalized storage facility for critical samples; supporting PepGen as BSO; managing CDC/USFWS permits and critical sample shipments. Supervising two scientists (one PhD, one Master's) on a daily basis to design/plan preclinical and clinical activities to support clinical trials for two neuromuscular indications.

Bioanalytical support of a commercial biologic and human monoclonal antibody (published): Developed and qualified Complement C1-INH, C4 and C1q ELISA assays and performed human plasma sample analysis to quantitatively detect exploratory biomarkers (PD marker); transferred assays to CRO. Developed and performed ligand binding assays (ELISA, MSD) to support an ongoing clinical trial. Provided bioanalytical support for Pre-filled syringe vs Auto-injector project (clinical) by managing PK/ADA assays; supported preclinical complement pathway regulation project. Developed Lateral Flow Assay as a diagnostic tool to detect Hereditary angioedema (published). Reviewed test protocols and qualification/validation reports in support of clinical programs. Supported PD marker assay development/validation for an ongoing clinical trial and an external exploratory biomarker project. Managed a team of scientists (8), conducted performance evaluation, screened resumes, and conducted interviews to provide hiring support to multiple departments i.e., Research/DMPK/Translational/Gene Therapy-Analytics and Vector production/CLD/Operations.

Studied anti-inflammatory effects of alpha7 ligands on NFKB-mediated inflammatory signaling pathways. Developed immune and non-immune cell-based quantitative assays to measure cytokine-induced biomarkers. Compiled and analyzed data, wrote the manuscript and submitted to scientific journals for publication.

Evaluated small molecules' prophylactic and therapeutic effects in multiple sclerosis using murine experimental autoimmune encephalomyelitis (EAE) model: Isolated lymph nodes and spleen from EAE mice; measured drugs' effects on MOG (35-55) treated splenocytes by lymphocyte proliferation assay; cytokines measured by BD cytometric bead array. Investigated the effects of small molecules in spontaneous systemic lupus erythematosus (SLE) model and DSS-induced colitis model; weighing and dosing animals, scoring based on disease severity, measuring proteinuria scores in SLE mice.

Optimized drug discovery methods to evaluate the effectiveness of hit or lead compounds in modulating Cystic Fibrosis Transmembrane Regulator (CFTR) protein surface expression: Improved western blotting and immunoprecipitation procedures to quantify CFTR protein (cystic fibrosis marker) levels to validate lead compounds. Conducted Ussing chamber (electrophysiology-based ligand binding assay) experiments on hit compounds to identify promising lead compounds using primary human lung cells. Collaborated with the medicinal chemistry team to develop hit or lead compound analogues. Analyzed data and presented/reported in company meetings; data recorded in electronic notebooks (ELN).

Standardized affinity reagents and robotic platforms to automate protein complexes purification (published): Expressed and purified more than fifteen Flag and HA-tagged proteins in mammalian cells using retroviral vector system. Established and compared manual and semi-automatic co-immunoprecipitation procedures using Thermo KingFisher 96 magnetic bead processor. Performed characterization of protein complexes by western blotting, SDS-PAGE and mass spectrometry. Examined protein complexes network caused by cancer causing viruses (published): Purified protein complexes by tandem affinity purification and performed quality control testing by Coomassie Blue staining and mass spectrometry. Studied 'Ku protein complex' composition, localization and its effect on DNA break ends (published): Isolated and purified affinity tagged Ku protein by anti-Flag/anti-HA beads from HeLa cells and performed proximity ligation assay to discover protein-protein interaction in vivo. Performed chromatin-IP (ChIP) to determine Ku protein-DNA interaction. Enhanced phosphopeptide fractionation for sharper and well separated peaks. Developed novel method for selective analysis of kinase phosphorylation sites using cancer cell lines and primary cells (published): Profiled kinases using western blot and HPLC. Optimized and developed methods to efficiently capture protein kinases from HeLa cells. Trained and supervised four NIH-funded CEGS program technicians.