Ching-Hung Chang

Supervised molecular virology core: plasmid cloning and lentivirus production for in vivo CAR-T.

• Designed and executed plasmid construction, and managed the plasmid bank.
• Maintained HEK adherent and suspension cells for producing LVV vectors, and established stable cell lines for personal research projects.
• Coordinated CROs to generate large plasmid prep for CMC, and performed a DNA quality check.
• Analyzed GOI expressions with the Quanteon cytometer and FlowJo.
• Explored new methods to shorten TAT, programming the epMotion liquid handler to automate the p24 ELISA assay, and using the Nanopore MinION sequencer to screen plasmid clones.

Engineered oncolytic RNA virus to effectively produce target proteins.

• Constantly conducted plasmid construction to mutate RNA virus, in vitro transcription, and RNA purification to produce vRNA, and transfected cells to produce viruses (full-length vRNA product produced without degradation most of the time).
• Checked the titers and reporter expressions by plaque assay and IncuCyte cell scanning.
• Screened regions in the genome of Coxsackievirus A21 and found a spot where the inserted Lucia luciferase gene could have 7X higher luciferase activities than no insert.

Set up the lab during COVID-19 pandemic and produced data for making go-no-go decisions

• Cultured insect SF9 cells for producing baculoviruses, verified protein expressions post virus transductions by Western blotting and measured nuclease-resistant copy number of virus by qPCR
• Determined baculovirus titers by flow-based quantifying gp64+ (PE) percentage of transduced cells

Researched the interactions between innate immunity and gene therapy vectors.

• Routinely cultured immune cell lines and human primary liver cells (from Lonza).
• Ordered compounds (FDA-approved drugs) are dissolved in appropriate solvents to make inventories.
• Supported projects in formulation and molecular biology groups.
• Patent (US WO2021030312: Methods and Composition for Reducing Gene or Nucleic Acid Therapy-Related Immune Responses) with the use of reporter cells, 96-well cell imaging, cell viability/toxicity assays, and Prism.

Supported baculovirus genome engineering.

• Characterized recombinant baculovirus genomes (130 kb) by restriction fingerprinting.
• Set up qPCR protocols for estimating Rhabdovirus levels in the insect cells after drug treatment.
• Completed the sequencing of a plasmid using NGS methods, the Nextera Flex kit to construct libraries, and then SPAdes software to perform in silico de novo assembly.

Contributed to lab operations focused on AAV gene therapy.

• Established an effective way to tell the difference between homozygous and heterozygous genotypes.
• Set up the AAV NAb assay, established reference serum from mixed blood of healthy donors, carried out reporter virus transduction, and the following quantification by flow cytometry.

Assisted in managing various projects.

• Successfully engineered a human erythroblast cell line using gene editing technology and characterized it by T7E1 assay, Sanger sequencing, and flow cytometry.
• Significantly improved the primary culture method of amplifying PGC (chicken primordial germ cells) to make them visible after 21 days instead of 35 days post-isolation of blood cells from hatched eggs.
• Validated commercial complement factor ELISA kits with mouse serum and plasma samples.

Oversaw Hiseq operations while supervising QPCR teams.

• Managed sequencing projects by effectively interacting with the lab manager, as well as the library preparation, QPCR, and Hiseq groups.
• Prepared laboratory documentation following CAP regulations.
• Solved technical problems, including quantification and sequencing issues.
• Handled library quantification, pooling plans of barcoded libraries, and subsequent cBot loading concentration.
• Operated Illumina sequencer, HiSeq 2000/4000, for next-generation sequencing.
• Established a novel mathematical way to replace bad ST curves with a previously good one in the process of qPCR data, avoiding the non-necessary repetition of whole 96-well plate reactions.