• Successfully produced multiple monoclonal antibody candidates for therapeutic immune checkpoint targets using hybridoma efusion technology• Designed, validated and carried out the detailed operational processes and experiments for animal immunizations, hybridoma screenings by ELISA or in-cell ELISA, antibody validation by Flow cytometry, cell-based functional assays and off-rate ranking (Octet), monoclonal antibody characterizations for cell surface and soluble protein binding properties, including binding affinities, using ELISA, Flow cytometry, kinetics by Octet, and cell-based functional blocking assays• Developed antibody purification methodology and cell-based functional assays for hybridoma screening and Ab development; Developed mammalian cell lines for Ab validations• Successfully delivered multiple CRO hybridoma technology-based antibody generation and antibody development projects. Assisted other CRO projects with AKTA purification of mAbs, and customer relations

Molecular Biology, Antibody Development and Validation• Established and optimized transfection and cell culture conditions of the mammalian expression system for recombinant antibodies using shake flasks and spinner flasks• Expressed and purified recombinant antibodies with yield up to grams per liter• Authored the SOP of mammalian expression for recombinant antibodies. Produced the first 11 in-house antibodies as a proof of concept for dramatic increase of yield and quality• Transferred the mammalian expression technology for recombinant antibody to Department of Production. Trained fellow scientists for using the technology platform • Developed and validated over 50 antibody products. Released products through effective team effort and collaborations from various Departments. • Conducted practices in compliance with ISO 9001• Provided scientific and technical support to the Production Department for the released antibody products. Offered trouble-shooting advice to customers on antibody-based applications• Expressed, purified and characterized high quality recombinant cytokines and other mammalian antigen proteins using biochemical and cell biological methodologies• Developed and optimized siRNA technology in various mammalian systems with varying reagents and conditions to knock-down protein expression to validate antibody specificity• Established and optimized the baculovirus expression and purification system for highly active kinase productions. Provided kinases and kinase mutants, e.g., EGFR and EGFR mutants products for drug screening and research studies

• Characterized the signaling pathways mediated by angiotensin and bradykinin receptors • Examined properties of the G-protein receptors using chimera and mutant receptor constructs in mammalian cells, real time PCR, oligodeoxynucleotide microarrays and phospho-specific protein screening technologies

Protein Purification• Produced with high efficiency the functional, purified protein kinases (5 – 10 milligrams with 90% purity) for HTS screening (stages I and II) for drug discovery• Formulated the purified kinase proteins for optimal function under various conditions (e.g., salt, pH, temperature, kinetics, DMSO and glycerol)• Purified protein targets by FPLC, including protein kinases, G-protein coupled receptors and nuclear hormone receptors•Constructed expression vectors and carried out cell culture (HEK293T and baculovirus system) for target protein productions

• Designed and constructed expression vectors for the HDL receptor and other control receptors for the high level expression of the proteins using molecular biology techniques• Expressed the receptor in insect cells using baculovirus expression system as well as in bacteria. Optimized the infection and culture conditions to achieve high levels expression of the proteins• Expressed the recombinant receptors in mammalian cells using cell culture and cell transfection technologies. Established stable cell lines and selected clones based on high receptor activities• Purified the receptor from the whole cell lysates using antibody-affinity chromatography, and identified the purified receptor using silver staining and Western blot analysis• Developed a liposome-based ligand binding assay system for the receptor. Validated and utilized the assay for functional analysis of the receptors• Developed cell-based assays, e.g., enzyme-linked immunoabsorbent assays (ELISA), immunoprecipitation and cross-linking, and analyzed the function of the receptor using receptor binding assays and fluorescence microscopy, fluorescent-activated cell sorting (FACS), flow cytometry, ELISA, immunoprecepitation and cross-linking

• Cloned the first two chloroplast RNA polymerase sigma factor genes using genomic DNA cloning, rapid amplification of cDNA ends (RACE) and DNA sequencing techniques• Characterized the sigma factor genes using Southern blot, Northern blot, primer extension and RT-PCR analyses• Constructed expression vectors, overexpressed the recombinant sigma factor proteins in E. Coli and purified the proteins using the anti-HIS tag antibody-affinity chromatography.• Characterized the antibody generated against the pure recombinant protein• Isolated the crude and highly purified chloroplast RNA polymerase using gel filtration, ion-exchange, DNA affinity and antibody affinity chromatography and gel electrophoresis techniques• Characterized the enzyme activities using in vitro transcription assays, and functionally identified the putative sigma factor proteins with Western blot analyses and immunoprecipitation assays