Routine (weekly) screening of compounds, data analysis with Gene data, reporting data to chemists, supporting assay transfer to CROs, SOP writing, data QC in house and at CRO Enzyme activity assay development and characterization (fluorescence intensity, polarization, FRET, luminescence):Determination of enzyme reaction kinetics (enzyme linearity, substrate Km determination using initial velocity)Evaluation of plate uniformity and Z' for HTS screening QCRoutine determination of compound IC50 to evaluate enzyme inhibition and occasional determination of compound MOI using enzyme kinetics assay and binding assays (such as SPR)Characterization of protein-protein and protein-ligand /small molecule binding using SPR (steady state binding and kinetics), binding using DSF thermal shift assay, ITC Characterization of redox cycling activity in compounds using fluorescence and absorbance assays

Evaluation of protein expression and localisation (cytoplasm, membrane) using western blotting Generation of stable cell pools by FACS sortingAnalytical FACS to quantify membrane protein enrichment after transfectionCell based activity characterization [HTRF to measure cAMP, pERK]Optimization of membrane protein solubilization and purification ATPase activity assay using ADP-GLO luminescence assayMolecular cloning (site directed mutagenesis, subcloning inserts into expression vector)Development of PCR based assay to evaluate splice acceptor and donor variants on splicing of gene variants

Molecular cloning, expression, purification, and characterization of protein ligands and antibodies for cardiovascular, metabolic, and oncology therapeuticsMolecular cloning:-Using RT-PCR and PCR, sequenced antibodies from hybridoma mRNA-Cloned antibody constructs using restriction digestion and ligationExpression and Purification:-transiently transfected mammalian cells to express recombinant protein ligands and recombinant antibodies-purified proteins using affinity, metal ion, size exclusion, and ion exchange chromatography by gravity and AKTA FPLC systems-quantified endotoxin concentration by LAL method-performed quality control analysis by intact mass spectrometry (LC-MS QTOF), SDS-PAGE, HPLC [analytical size exclusion chromatography]Protein Characterization:-Calculated potency and efficacy of antibodies using cell based HTRF and AlphaLISA activity assays-Determined binding potency (EC50) of antibodies to heterologously expressed proteins on cells using MSD technology-Determined real-time kinetics of antibody-protein binding using SPR (Biacore) and BLI (Octet-ForteBio)-Binned antibodies into epitope groups using BLI (Octet-ForteBio)-Developed ELISA assay to quantify biomarker concentration in plasma-Determined half-time of protein therapeutics in vivo by pharmacokinetic (PK) analysis

-cloned recombinant collagen-like protein constructs to study collagen-enzyme/protein interactions-optimized the expression and affinity and ion exchange purification (AKTA FPLC) of recombinant collagen-like proteins in Escherichia coli-assessed the purity and folding of proteins using SDS-PAGE and Circular Dichroism Spectroscopy-determined melting temperature of proteins using Circular Dichroism Spectroscopy and Differential Scanning Calorimetry-performed ELISA to assess binding affinity of protein ligands to recombinant collagen-developed a fluorescence polarization assay to quantify binding affinity of protein-peptide interactions

-performed purification of antibodies and protein ligands using affinity, size exclusion, metal ion, and ion exchange chromatography under low endotoxin conditions using AKTA FPLC system-protein and antibody aggregation, purity, and function were assessed using analytical size exclusionchromatography, SDS-PAGE, ELISA and BLI-developed protocols for purifying recombinant antibodies using affinity and ion exchange chromatography-developed functional assays to quantify binding affinity of antibodies to antigen and assess level of bovine antibody contamination-maintained lab stocks, ordered reagents, and sterilized buffers and media using the autoclave

-created single amino acid mutant constructs using site directed mutagenesis -expressed recombinant proteins using Escherichia coli-purified the intracellular domain of hZIP4 using affinity chromatography -developed buffer forumlation for protein storage using dynamic light scattering to assess proteinaggregation-assessed protein purity by SDS-PAGE and western blot- performed isothermal titration calorimetry and fluorescence spectroscopy to characterize the binding affinity of the protein to zinc-developed a fluorescence based assay to quantify binding affinity of the protein to zinc-maintained lab stocks, ordered reagents, sterilized buffers and media using autoclave

-expressed recombinant proteins using SF9 insect cell and Escherichia coli expression systems-purified recombinant protein using affinity, metal ion, size exclusion, and ion exchange chromatography by gravity and AKTA FPLC system-assessed purity of protein using SDS-PAGE-maintained lab stocks, sterilized buffers and media using autoclave

taught swimming lessons as an instructor, performed cleaning and maintenance duties, and watched swimmers at the outdoor pool