Arthur Hauenstein Email & Phone Number

South San Francisco, California, United States

Biophysical characterization of lysosome targeting chimeras (LYTACs).

Brisbane, California, United States

Contributed to Infectious Disease and Vaccines (IDV) therapeutic pipeline, including proposing new targets and supporting existing programs as protein sciences subject matter expert (SME) in viral biochemistry group. Supervised 1 direct report for 3+ years expanding this individual’s role from RA II to SRA. Hired and managed 1 contractor (6 months) and 1 summer intern (12 weeks).o Developed new project from target validation to hit ID as TA lead on novel host-virus protein-protein interaction (PPI) target. Engaged key opinion leaders (KOLs), cross-functional teams, and senior leadership to gain portfolio entry approval. • Designed, expressed, and purified target proteins. Conducted target validation and MoA studies. • Worked with 20+ member, global, cross-functional team (including one direct report) to develop hit-finding plans and orthogonal assays (FRET, SPR, AlphaLISA, HTRF). Completed DEL, AS-MS (small-molecule), mRNA-display macrocyclic peptide screening campaigns through Hit ID stage. • Identified 2 hit series with different inhibition mechanisms that have nM to sub-nM potency in binding and biochemical assays (manuscript in preparation). • Communicated project milestones, status updates, and team recommendations to senior leadership and key stakeholders. o Supported multiple programs as a protein sciences SME within IDV • Expressed and purified recombinant proteins of interest from insect and bacterial cell hosts enabling affinity screening and downstream assay development. • Developed numerous biochemical (FRET-based activity assay) and biophysical (AlphaLISA-based PPI assay, MST, ITC, DLS, DSF) assays to support target validation and MoA studies.

Natick, Massachusetts, United States

Core member of seed-stage company founded by Hao Wu and Judy Lieberman (Boston Children's Hospital / Harvard Medical School). Collaborated on design and setup of laboratory, including CapEx purchasing and equipment maintenance. Contributed to strategic planning, including project prioritization, target / indication assessment and selection, and hit finding strategies. o Supported protein production and hit profiling on lead program.o Proposed and gained approval for new auto-inflammatory disease target by engaging scientific advisory board (SAB) members, consultants, and senior management.• Conducted target validation and MoA studies.• Conceptualized and performed focused, target-based screen (ADP-Glo assay, ~3K diverse small-molecule kinase library) including construct design, protein production, and tool finding.• Developed orthogonal SPR, NF-kB luciferase reporter, Western Blot, and ELISA-based assays.• Identified tool compound with low micromolar binding and biochemical activity.

Boston, Massachusetts, United States

Program in Cellular and Molecular Medicine (PI: Hao Wu, Ph.D.)Studied the activation and assembly mechanisms of innate immune complexes.o Project 1: Investigated the binding of polyubiquitin chains to NEMO• Developed in vitro synthesis and purification scheme to make distinct length K63-linked and M1-linked polyUb chains.• Conducted SEC-MALS and SEC-SAXS on full-length NEMO. • Identified regions of NEMO that allosterically regulate binding to M1-linked polyUb chains. Characterized NEMO binding preference for M1-linked over K63-linked polyUb chains.o Project 2: Characterized the effect of the diffuse-large B-cell lymphoma gain-of-function mutation L265P on MyD88 TIR domain structure.• Conducted extensive protein engineering and mutational analysis of TIR-TIR interaction interfaces to disrupt/limit L265P-driven TIR oligomerization for structural studies.• Performed in vitro reconstitution studies of TLR-MAL-MyD88 assembly on supported lipid bilayers.• Generated stable, doxycycline-inducible cellular assay in HEK293 to monitor MyD88 assembly by confocal fluorescence microscopy.o Project 3: Investigated NLRP3 inflammasome activation mechanisms.• Engineered, expressed, and purified various constructs of NLRP3 proteins from mammalian (HEK293-F) and insect cells (Sf9)• Designed fluorescence polarization (FP) assay to study NEK7 dependent NLRP3 activation.• Performed structure-guided mutational analysis of NLRP3 binding to its co-activator, NEK7, using pull-down assays and by functional reconstitution in NEK7 KO iBMDM cells.• Created clones of numerous functional, fluorescently labeled inflammasome pathway proteins in lentiviral vectors to generate stable cell lines for monitoring spatiotemporal dynamics of inflammasome activation by live-cell imaging.• Validated a novel regulator of NLRP3 inflammasome activation, HDAC6, by generating CRISPR/Cas9 genetic knockouts in mouse iBMDM cells and conducting functional, cellular reconstitution studies.

San Diego, California, United States

Department of Chemistry and Biochemistry (PI: Tom Huxford, Ph.D.)Conducted Structural/Biochemical/Biophysical Studies to Elucidate the Activation Mechanism and Substrate Selectivity of the Human Inhibitor of kappa B Kinase 2/Beta (IKK2/B).o Developed novel protein constructs and optimized baculoviral insect cell expression, purification, and crystallization conditions that led to the determination of the first human inhibitor of kB kinase 2 (IKK2) structure by X-ray crystallography in collaboration with the Gourisankar Ghosh lab (UCSD).o Elucidated the role of higher-order oligomerization in the trans-autoactivation mechanism of IKK2 by a variety of biophysical and biochemical techniques including size-exclusion chromatography-multi-angle light scattering (SEC-MALS), small-angle x-ray scattering (SAXS), analytical ultracentrifugation, and in vitro kinase assays (32P autoradiography).o Characterized the minimal determinants of oligomerization-dependent autophosphorylation and substrate selectivity through protein engineering and mutational analysis studies of a monomeric IKK2 construct.

Doctor Of Philosophy (Phd), Biochemistry

Doctor Of Philosophy (Phd), Biochemistry