Process Development

Virendra Pratap Singh

Email: *******@****.***, Phone:410-***-****, 4812 Eldon Green CT, Halethorpe MD-21227

Linkedin: http://www.linkedin.com/pub/virendra-singh/37/295/4b8

Objective

To obtain a position in bioprocess/chemical industry where I can effectively utilize my technical skills and contribute in the growth of an organization.

Professional Summary

Over 5 years of full time research and development/process development experience in recombinant protein purification, filtration and fermentation in industry.

Technical Skills

Chromatography Lab to process scale chromatography (BPG 450/750), operation and column packing of reverse phase, ion exchange, size exclusion, hydrophobic, affinity and mix mode chromatography.

Scale up of chromatography. ( AKTA) prime, explorer, pilot and process.

Analytical Techniques Characterization of proteins by SDS-PAGE, Isoelectric-focusing, Western blotting, 2D gel electrophoresis and analytical HPLC. Agarose gel electrophoresis, PCR, Capillary zone electrophoresis.

Protein quantification with HPLC, UV spectroscopy and chemical methods like Lowry’s and Bradford.

Filtration Tangential flow filtration, sterile filtration and dead end flow process optimization, throughput analysis. Filtration scale up.

Filter Validation Sterile filter validation by bacteria challenge test, chemical compatibility and integrity testing, burst pressure analysis.

Fermentation SIP,CIP and operation of fermentation from 2L to 300L,sterile handling, scale up.

Others Design of experiments (DOE), Protein refolding, lyophilisation and centrifugation

Qualification SOPs preparation and qualification of equipments.

Education

• Pursuing MS program in Chemical Engineering at University of Maryland Baltimore County USA. Expected May 2012.

• Master of Technology (Biotechnology) Anna University India, First class with distinction. May 2005.

• Bachelor of Engineering (Bio-Chemical Engineering) Kumaun University, India, First-Class. June 2003.

Professional Experience

Project Leader (Downstream Processing), Apr 2008-Jul 2010, Sartorius-Stedim Biotech (R&D), Bangalore

• Worked on dynamic binding capacity determination and flux determination of membrane adsorber chromatography ( anion, cation exchange) capsules in various buffer systems using Bovine serum albumin and Lysozyme.

• Throughput analysis of the cartridge and disk filters made of various membranes like polyether sulfone (PESU), cellulose acetate (CA) and polypropylene (PP) of different pore sizes in various combinations.

• Sterile grade filter validation by bacteria challenge test, product specific integrity testing and physical/chemical compatibility determination of membrane filters in various biochemical solutions.

• Scale up/filter sizing of dead end filtration systems based on cake formation, gradual pore plugging or complete pore plugging model.

• Cross flow filtration optimization for various recombinant human therapeutic proteins in various buffers and scale up.

• Chemical compatibility analysis of new generation PESU membrane filters in various organic, inorganic, salts, acids and base solutions to study the effect of chemicals on the integrity and physical strength of the membrane, intended for sterile filtration.

• Lab setup, qualification of equipments and SOPs preparation.

Research Associate, Jun 2005–Mar 2008, Wockhardt Research Centre Aurangabad, India

• Purification process development (HCP, Dimer, Isoforms and endotoxins separation as major considerations) and scale up of rh-Erythropoietin by using affinity (blue sepharose), reverse phase (source 30 RPC), anion exchange (Q-sephrose) and gel filtration chromatography (superdex G-200).Scale up from lab to process level.

• Optimization of GCSF to PEG-GCSF conversion and its purification process standardization using cation exchange (SP sepharose) chromatography.

• Development of solubilisation and refolding of therapeutic proteins like rh-GCSF, rh-Glargine (insulin analogue), and rh-IFN beta 1b expressed as inclusion body in various strains of E. coli.

• Optimization of conversion process from pro-insulin expressed in H. polymorpha to insulin-ester.

• Involved in purification development of insulin from (1) Conversion of pro-insulin to insulin-ester (transpeptidaion reaction by using trypsin); (2) Organic precipitation (isopropanol); (3) Ester purification (amberchrome CG-300 M); (4) insulin-ester hydrolysis (triflouroacetic acid) (5) RPHPLC (XT-30), Scale up of the process and technology transfer from R&D to process scale.

• Experience of working in cGMP(current good manufacturing practices) facility and also attended academic courses for cGMP and regulatory affairs

Certificate on Sterile filter Validation

Sartorius-Stedim Biotech (R&D), Goettingen Germany, Jul – Aug 2008

Bacteria challenge test by direct/indirect method to validate sterile grade filters, physical/chemical compatibility of the membranes, burst pressure analysis, product specific integrity testing and throughput analysis of various filters.

Clamping force analysis and defect determination in the cross flow cassettes by smoke test.

M.Tech Project

‘Large scale production of 'pVAX-ALT2' for DNA vaccine development of Lymphatic Filariasis’, a disease caused by mosquito-borne Brugia malayi. Fermentation was carried out in 5 liter scale by cultivating E.coli (TOP 10) in glucose –yeast extract medium. Recombinant plasmid was recovered by alkaline cell lysis followed by purification with anion exchange chromatography (Q-sepharose).

MS (Thesis)

Cell free expression and purification of sensor protein at small scale.

Achievements

• Received (DBT-New Delhi) fellowship during M.Tech (2003-2005).

• Received merit scholarship during B.E.(1999-2003)

• Qualified All India ‘DBT-JRF' examination in 2005.

• Qualified 'Integrated Scholarship Examination' in 1992 and 1995 conducted by UP Board Allahabad and received merit scholarship from class 6th to intermediate level.

• Was selected through campus interview conducted by Wockhardt (Biotech R&D) for its research centre at Aurangabad, India.

References

Dr. Antonio Moreira

Professor and Vice Provost

University of Maryland Baltimore County, MD 21250

Email: *******@****.***

Phone: 410-***-****

Dr. Vinod Kuberkar

Head-Business Development

Intas Biopharmaceticals

Ahmedabad, India

Email:*****.********@**************.**.**

Phone: +91-972*******

Dr. Douglas Frey

Professor and Graduate Program Director

University of Maryland Baltimore County, MD 21250

Email: ******@****.***

Phone: 410-***-****

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