BRIAN NELSON

Assay Development

• Assay development for test methods designed to be fit for purpose (gene identity testing, gene copy number, gene linkage, etc) for QC transfer
• Optimized drug product characterization by Western blot for capsid/viral protein detection
• Drafted, released, and redlined test method protocols and method development reports

QC Control

• Mycoplasma testing of cell culture, cell banks, final drug product, etc
• Physical Titer quantitation by qPCR for drug product, IPT, and research samples from RNA column extraction
• ID testing for different vector constructs to confirm product identity
• Residual host cell DNA (hcDNA) testing
• Deviation report experience via impact assessments, root cause analysis, and discovery
• Routine testing for samples for stability, in-process testing, engineering runs, and GMP

• Knock-out and knock-in designs for β-Thalassemia genotype with primer design, locus targeting, sgRNA optimization, etc
• Next Generation Sequencing (NGS) using CHANGE-Seq , CIRCLE-Seq , GUIDE-Seq , rhAMP-Seq , and Nextera -based sequencing (MiniSeq)
• ddPCR to quantify knock-in events, INDELs, expression, and AAV infection efficiency
• cDNA preparation of RNA from mouse/monkey tissue/cells
• RT-qPCR to quantify knock-in expression levels of the HBB gene using SYBR green dye
• BCA and Western Blot assays to quantify target SaCas9 and KI products in mice
• Preparation of liver tissue samples
• ELISA to quantify Factor VIII expression for Hemophilia A research for gene therapy and gene editing
• Transformation of E. coli using Giga, Maxi, and Mini prep assays for isolation of plasmid constructs
• In-fusion cloning to create SaCas9 variants that include codon-optimized constructs
• Fluorescence-activated cell sorting (FACS) of cells by cytofixation and intracellular staining

• DNA extraction from trophoblasts/blastomeres derived from inseminated embryos in vitro from IVF clinics and PCR
• Identified losses, gains, deletions, duplications, inversions, translocations, and chromosome mosaicism through karyotype analysis
• MiSeq - operation, maintenance, troubleshooting
• (WGS) Whole Genome Sequencing of DNA from D3-D6 IVF embryos
• Biopsy reports/networking with IVF clinics
• PGD testing and Mitograde DNA testing
• Microarray (aCGH) post-hybridization processing and data analysis

• Conducted luminescence and absorbance ELISA for illicit and prescription drugs from collected hair samples
• GCMS for hair sample positives from initial screening
• Detected by confirmation of the presence of targeted analytes and metabolites by antibodies from ELISA assays