Naveenkumar Singh, Ph.D.
Downstream Process Development

Naveenkumar Singh received his B.Sc. in Biotechnology (2007) at University of Pune (Pune, India), M.Sc. in Biotechnology (2009) at Jaipur National University (Jaipur, India), and Ph.D. in Biochemical Engineering (2016) at Jacobs University Bremen (Bremen, Germany).

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Ph.D. in Biochemical Engineering, with 6+ years relevant experience and a thorough knowledge of the downstream process.

During his Ph.D., Naveen Kumar Singh worked under the supervision of Prof. Marcelo Fernández-Lahore. His research dealt with designing experiments for developing fiber-based and cryogel-based adsorbents for capturing large therapeutic biomolecules (proteins, plasmids, and monoclonal antibodies). He had successfully evaluated the in-house fiber- and cryogel-based chromatographic adsorbents with the commercially available adsorbents and the in-house adsorbents had shown similar or higher productivities compared to the commercial adsorbents.

 

In February 2017, Naveen Kumar Singh joined the group of Prof. Merlin L. Bruening in the Department of Chemical and Biomolecular Engineering at the University of Notre Dame as a Postdoctoral Research Associate. His current research focuses on developing novel bioseparation processes by introducing polyelectrolyte multilayer films onto membranes/monoliths for the purification of biotherapeutics like monoclonal antibodies.

 

Skilled in both protein purification for R&D purposes, as well as purification scale-up process development for technology transfer to manufacturing. Highly versed and polished in the execution of downstream multi-steps purification of novel therapeutic proteins to support early phase clinical trials studies. Strong background in the development of protein purification protocols (AKTA-based protein chromatography) at small and larger scale. An AKTA/UNICORN expert across all models and software versions. Experienced with protein analytics, UF/DF, filtration, and formulation.

 

Specialties:

  • Recombinant proteins, monoclonal antibodies, and nucleic acid (pDNA and mRNA) purification
  • Different modes of chromatography
  • Bead-, fiber-, monoliths- and membrane-based adsorbents
  • Synthesis and evaluation of ion-exchangers, and affinity-based adsorbents
  • Basic analytical techniques for protein and nucleic acid (HPLC analysis, protein and nucleic acid quantification methods, electrophoresis, etc.)
  • Mass spectrometry characterization of therapeutic antibodies and other recombinant proteins
  • Scale-up /-down
  • Strong background in the development of protein purification protocols (protein chromatography) at small and larger scale
  • Working knowledge of good laboratory practices and cGMP
  • Column Packing
  • Tangential flow filtration (TFF), Ultrafiltration, Diafiltration
Experience

Feb. 2017 – Present

Postdoctoral Research Associate
University of Notre Dame
  • Developed porous membranes and monoliths for rapidly capturing monoclonal antibodies.
  • Successfully developed/evaluated enzymatic deglycosylation of monoclonal antibody using LC-MS.
  • Extensive hands-on skills in middle down/bottom up approach for peptide mapping, protein sequences identification, and protein posttranslational modifications (PTMs) characterization using mass spectrometry.

May 2016 – Oct. 2016

Research Associate (Postdoc)
Jacobs University Bremen, Bremen, Germany
  • Tailored the gPore fiber-based system in the recovery and purification of monoclonal antibodies.
  • Scaled a chromatography method (1 mL to 250 mL) for monoclonal antibodies purification.

Jun. 2011 – Apr. 2015

Research Associate (Downstream Processing)
Jacobs University Bremen, Bremen, Germany
  • Developed, modified, and evaluated the performance of gPore fiber-based materials, which can be functionalized and used to purify a range of bioproducts efficiently.
  • Successfully designed a single step downstream process for B-phycoerythrin purification and scaled it from 1 mL to 250 mL chromatographic column.
  • Established lab-scale chromatography methods for recombinant proteins, monoclonal antibodies, pDNA, and mRNA purification.
  • Trained 6 undergraduate, master, and Ph.D. students for the conduct of independent research.

Jul. 2010 – Jan. 2011

Trainee and Executive Trainee (Downstream Processing)
Wockhardt Ltd, Aurangabad (M.S.), India
  • Performed large-scale purification of recombinant insulin glargine under cGMP for clinical trials.
  • Carried out solubilization of inclusion bodies, refolding and final conversion of proinsulin to insulin.
  • Operated different column operations (Ion-, buffer-, cation-exchange chromatography, and RP-HPLC).
  • Assisted project manager in shortlisting better trypsin enzyme by running a couple of experiments.
  • Followed and maintained cGMP, standard operating procedures, health, and safety policies.

Education

2011–2016

Ph.D. in Biochemical Engineering
Jacobs University, Bremen, Germany

Thesis: “Preparation and characterization of grafted adsorbents and their applications in biomolecule purification”.

Supervisor: Prof. Dr. Marcelo Fernández-Lahore.

Within this project, Hybrid Disposable Cartridges (HDC), a hybrid approach created by merging existing textile technology with Material/polymer chemistry was used to generate a cost-effective and disposable system supporting eco-friendly bioprocessing.

The goal here was to evaluate the performance of gPore NW materials, which can be functionalized and used to purify a range of bioproducts efficiently.

The further objective was to tailor the gPore NW system, which has already shown excellent results in the recovery and purification of natural as well as recombinant proteins, to other products, like pDNA or monoclonal antibodies.

2007–2009

M.Sc. in Biotechnology
Jaipur National University, Jaipur, Rajasthan, India

Thesis: “Regulation of T cell immune responses by chemokines CCL19 and CCL21”.

Supervisor: Dr. Bhaskar Saha, National Centre for Cell Science, Pune.

2004–2007

B.Sc. in Biotechnology
University of Pune, Pune, Maharashtra, India
Training

July 2009 – Sept. 2009

Trainee
ILS Bioservices, Gurgaon, Haryana, India

3-month foundation certified program in Genomics.

Conferences and Workshops
Talks at Conferences
  1. 2012 5th Monolith Summer School & Symposium – MSS2012, Slovenia.
  2. 2007 Modern Approaches and Innovations in Biotechnology and Biodiversity conservation, India.

Posters at Conferences
  1. 2016 European Society for Biochemical Engineering Sciences (ESBES) 2016, Ireland.
  2. 2014 16th European Congress on Biotechnology, Scotland.

Workshops
  1. 2016 “QbD Related Modelling and Control Strategies” at the ESBES 2016, Ireland.
  2. 2012 “Purification of Biomolecules and scale-up” at the MSS2012, Slovenia.

Publications

N. K. Singh, R. N. Dsouza, M. L. Sánchez, S. Verma, E. Achilli, R. R. Vennapusa, M. Grasselli, and M. Fernández-Lahore, “Preparation and Characterization of Grafted Cellulosic Fibers and their Applications in Protein Purification“, Sep. Purif. Technol., 143 (2015) pp. 177–183.

N. K. Singh, R. N. Dsouza, N. S. Bibi, and M. Fernández-Lahore, “Direct Capture of His6-tagged Proteins Using Megaporous Cryogels Developed for Metal-ion Affinity Chromatography“, inAffinity Chromatography (Ed.: S. Reichelt), Spinger, New York, USA, Methods in Molecular Biology, 1286 (2015) pp. 201–212.

N. K. Singh, R. N. Dsouza, M. Grasselli, and M. Fernández-Lahore, “Gamma ray mediated functionalization of monolithic cryogels for macro-biomolecule purification“, N. Biotechnol., 31, Supplement (2014) pp. S127.

N. K. Singh, R. N. Dsouza, M. Grasselli, and M. Fernández-Lahore, “High capacity cryogel-type adsorbents for protein purification“, J. Chromatogr. A, 1355 (2014) pp. 143–148.

N. S. Bibi, N. K. Singh, R. N. Dsouza, M. Aasim, and M. Fernández-Lahore, “Synthesis and performance of megaporous immobilized metal-ion affinity cryogels for recombinant protein capture and purification“, J. Chromatogr. A, 1272 (2013) pp. 145–149.

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Keywords:

Process development, downstream purification process, downstream purification, downstream, recombinant protein, mAb, vaccines, vaccine, mRNA vaccine, DNA vaccine, scientist, centrifugation, filtration, pilot-scale, bench-scale, pilot scale, bench scale, laboratory maintenance, biochemistry, biology, biochemical engineering, chromatography, tangential flow filtration, flow filtration, tangential flow, AKTA chromatography, AKTA, Pilot, cross flow filtration, JMP software, JMP, shake flask, t flask, fplc, fast protein liquid chromatography

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Technical Competencies

Downstream Processing

Preparative chromatography (monoliths-, membrane-, and bead-based adsorbents)(ion-exchange, affinity, multimodal and hydrophobic-interactions), size-exclusion, reversed-phase chromatography, tangential flow filtration (TFF), ultrafiltration, diafiltration, centrifugation, HPLC.

Analytical Techniques

SDS-PAGE, Native PAGE, Western blotting, Southern blotting, Northern blotting, isoelectric focusing, spectrophotometry, confocal microscopy, ELISA, and ExperionTMautomated electrophoresis system.

Molecular Biology

Nucleic acid isolation and processing, cloning, reverse transcriptase-PCR.

Cell Culture

Bacterial cell culture, primary cultures (Murine Macrophages, T cells, Dendritic cells, Leishmania, etc.) and cell lines.

Fermentation

Batch and Fed-Batch fermentation (upto 500 L).

Instruments

ÄKTApilot/ÄKTAexplorer/ÄKTA FPLC (AKTA systems) and UNICORN software; FEI Magellan 400 (SEM); Dionex Ultimate 3000 (HPLC); Thermo Scientific LTQ Orbitrap Velos (Mass Spectrometer); Bruker micrOTOF-Q II (Mass Spec); Nicolet 6700 FT-IR spectrometer; alpha-SE Ellipsometer

Experience in GLP, cGMP, standard operating procedures (SOP’s), health and safety documentation.

Operating system (Computer)

Linux, MacOS and Windows.

Publishing (Computer)

html, LATEX, Microsoft Office, EndNote, Adobe Illustrator, Adobe Photoshop.

Scientific (Computer)

Origin, ChemBioOffice, MODDE, BLAST, FASTA, PROSITE, UNICORN.

Languages (Computer)

C, C++.

Website Designing (Computer)

Blogger, WordPress, Microsoft WebMatrix.

English (Languages)
Hindi (Languages)
Marathi (Languages)
German (Languages)
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