Expert Details
Protein aggregation, characterization, purification
ID: 732518
Massachusetts, USA
Keywords: Process development, DSP, Downstream processes, Gene therapy, Oncolytic Immunotherapy, AAV, Adenovirus, Lentivirus, enveloped-virus, viral vector purification, vaccine characterization, protein characterization, analytical characterization, antibody purification, biologics, Depth filtration, TFF, Tangential flow filtration, UF/DF, Chromatography, Vaccine, Covid-19, HPLC (IEX, SEC), RP-HPLC, AKTA FPLC, TFF, dynamic light scattering (DLS), DSC, CD, AUC, protein chemistry, biochemistry, NMR spectroscopy, SAXS, protein engineering, UV-vis, fluorescence spectroscopy, gel electrophoresis, biochemical, biophysical characterization, Fluorescence biophysical characterization, data analysis
SUMMARY
• Innovative protein biophysicist and biochemist with 10+ years of proven expertise in protein biochemistry and biophysics
• Led 150 + projects and worked in collaborative and multidisciplinary teams within academic and industrial set-ups
• Performed and supervised projects involving protein and antibody purification and characterization
• Interacted with teams within and outside the company to prepare statement of work, reports, expanded business opportunities in a one-to-one or team settings
• Prepared cost-benefit analysis on an in-house project to improve efficiency and curb losses in a long run
SKILLS
• Nucleic acids: Isolation of mRNA from mouse brain, isolation of plasmid DNA from E. coli DH5 strain, reverse transcription of mRNA, Polymerase Chain Reaction (PCR), Molecular cloning
• Recombinant proteins: Expression and purification of proteins such as prion protein and its variant, αB-crystallin, α-synuclein, 2-microglobulin, CRABP1 and its variants using bacterial expression system, 15N and 13C isotope labeling of proteins, protein modification using fluorophore labeling, SDS-PAGE, Western blotting, and Blue-Native PAGE
• Protein purification: Affinity chromatography, anion and cation exchange chromatography (CEX), HIC, preparative SEC, AKTA FPLC using custom packed columns including XKs (16, 26 and 50) and BPG100 columns, and Tangential flow filtration.
• Protein analysis: Reversed-phase-, and Size exclusion- HPLC (Agilent), LAL assay, Octet (titer assay)
• Spectroscopy: Nuclear Magnetic Resonance (NMR) (Varian 700), UV-Visible Spectroscopy, Fluorescence Spectroscopy (Anisotropy, FRET), Circular Dichroism (CD), Dynamic Light Scattering (DLS), Time Correlated Single Photon Counting (TCSPC), Isothermal Calorimetry (ITC), Small-angle X-ray Scattering (SAXS), Analytical Ultracentrifugation (AUC)
• Microscopy: Total Internal Reflection Fluorescence Microscopy (TIRF-M), Transmission Electron Microscopy (TEM), Atomic Force Microscopy (AFM)
• Productivity: Microsoft Word, Excel, and Power point, Origin, and SigmaPlot
• Programming: MATLAB, R, Python
Education
| Year | Degree | Subject | Institution |
|---|---|---|---|
| Year: 2010 | Degree: Ph.D. | Subject: Biochemistry | Institution: CCMB, India |
| Year: 2003 | Degree: M.Sc. | Subject: Life Sciences | Institution: JNU, India |
| Year: 2001 | Degree: B.Sc. | Subject: Agricultural Sciences | Institution: BHU, India |
Work History
| Years | Employer | Title | Department |
|---|---|---|---|
| Years: 2021 to Present | Employer: Undisclosed | Title: Head of Downstream Manufacturing | Department: |
Responsibilities:Leads a team of downstream scientists to develop scalable downstream processes for viral vectors for use in Gene, Cell and Oncolytic therapies and for Vaccine production and for therapeutics proteins and antibodies. |
|||
| Years | Employer | Title | Department |
| Years: 2018 to 2021 | Employer: Pall Corporation | Title: Senior Purification Scientist | Department: |
Responsibilities:Research |
|||
| Years | Employer | Title | Department |
| Years: 2016 to 2018 | Employer: LakePharma, Inc. | Title: Senior Scientist- Protein Sciences | Department: |
Responsibilities:• Led team as a project manager• Team lead-scale-up purification • Led as a Study Director for a few projects with 10-15 team members • Performed 150+ projects to their completion in one of the highest performing groups • Worked in the downstream process development (DSP) such as purification of antibodies and mammalian cells at milligram to gram scales under low endotoxin condition • Involved in purification and characterization of proteins from bacterial, yeast, insects and mammalian cells. • Performed IMAC, IEX, affinity including GST, streptactin, softlink, ProA purifications, membrane enrichments using N2 bomb and sonication • Performed SEC using S400, S200, S75 columns • Performed large scale purification using BPG100 columns with 1-2 L of resins • Characterized protein using aSEC, RP, CEX HPLC, DLS, CD • Designed and executed refolding scouts to find optimal conditions for refolding • Custom packed columns with bed volumes ranging from ml to L scale • Performed ultrafiltration and diafiltration using TFF for up to 50 L scale • Performed high through put protein purification • Extensively worked on Akta Start, explorer, FPLC and Pilot • Developed SOPs and prepared detailed report • Trained and supervised direct report • Trained and supervised new and old hires on the purification and characterization techniques including DLS, HPLC, TFF, custom resin packing, and Akta explorer • Extensive client interactions to prepare statement of works, reports, and expand business opportunities in a one-to-one or team settings • Handled multiple projects and client interactions simultaneously • Collaborated with various teams for the completion of projects • Directly involved in installation or training in new techniques such as DLS, Octet, and UPLC |
|||
| Years | Employer | Title | Department |
| Years: 2011 to 2016 | Employer: University of Massachusetts | Title: Postdoctoral Researcher | Department: |
Responsibilities:• Investigated crosstalk between folding and aggregation of a model protein and its engineered variants• Performed 15+ protein enrichments under minimal conditions to isotopically label protein • Learned and performed 1D, 2D HSQC, 3D HNCO, HNCA, HNcoCA, HNcaCO, CBCAcoNH, and HNCACB experiments • Processed NMR spectra and assigned protein under denatured condition • Performed non-uniform sampling to reduce NMR time • Developed a method using principal component analysis to investigate structural content of the unfolded state • Developed skills for R, Python and MATLAB programming • Performed protein folding experiments using CD, fluorescence measurements • Identified that the folding information is present in the unfolded state • Presented work in conferences and annual retreats • Trained and supervised high school student and undergrads |
|||
| Years | Employer | Title | Department |
| Years: 2010 to 2011 | Employer: Center for Cellular and Molecular Biology | Title: Research Associate | Department: |
Responsibilities:• Designed software for photo-acoustic spectroscopy (PAS) using LabVIEW• Performed stability analysis on recombinant humanized monoclonal antibody, Bevacizumab (Avastin, Genentech/Roche) using reverse phase-HPLC (RP-HPLC) |
|||
| Years | Employer | Title | Department |
| Years: 2004 to 2010 | Employer: Center for Cellular and Molecular Biology | Title: Graduate Research Assistant | Department: |
Responsibilities:• Studied misfolding and aggregation of mammalian prion protein• Cloned, expressed and purified mammalian prion protein and its variants • Discovered novel tertiary interactions between the N- and C-terminal regions of prion protein upon copper-binding using CD, fluorescence, SAXS, ITC, AUC, DLS, and NMR • Identified loss of nucleation as a cause for inhibition of amyloid formation of UV-exposed prion protein using CD, fluorescence, AFM, and EM • Authored three scientific papers and a book chapter • Presented various professional seminars on prion diseases • Trained and supervised high school students, undergrads and graduate students |
|||
Career Accomplishments
| Awards / Recognition |
|---|
| HONORS AND AWARDS • Best oral presentation award, in National Symposium on Biophysics-Biophysics in Medicine and Biology, organized by Indian Biophysical Society, India • Young Scientist Bursary to attend 63rd Harden Conference – Protein folding and assembly in vitro and in vivo, Ambleside, UK, organized by Biochemical Society • Senior Research Fellowship, Council of Scientific and Industrial Research (CSIR), India • Junior Research Fellowship, CSIR, India |
Additional Experience
| Other Relevant Experience |
|---|
| ADDITIONAL INFO • Workshop attended- Tips & Tricks: ACQUITY H-Class and QDa at Waters regional office • Was point-of-contact with service engineers for several biophysical instruments such as ITC, CD, DLS, and time-resolved fluorometers • Installed Endosafe Nexgen PTS reader with the remote help of application scientist |