Muralidhar Reddivari
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Experienced manager in therapeutic protein and virus expression and purification, bioprocess development including management of upstream and downstream process development and unit operations.
Experience
Reddivari Muralidhar, PhD
570 Winding Valley Cove, Collierville, TN, 38017, USA
Murali59_60002@yahoo.com
9013783224
Experienced manager in therapeutic protein and virus expression and purification, bioprocess development including management of upstream and downstream process development and unit operations.
Professional Summary
- Over 20 years of experience in adeno associated virus / lentivirus / recombinant protein expression and purification using mammalian cells, insect cells, yeast and bacteria.
- Upstream Process development - Maintain and run SS Bioreactors / Fermentors from 10L to 200L, SUB wave-bioreactors up to 200L.
- Harvesting techniques including depth filtration, TFF and smart flow filtration.
- Downstream Process development - Maintain and run protein purification platforms including AKTA Ready, AKTA AVANT, AKTA Pure and AKTA purifiers. Bio-Rad NGC purification platforms: development of purification strategies based on protein chemistry.
- Fill/finish operations using ChaseLogeman / Bausch and Stroebel and SKAN isolators
- Manage and train a team of scientists / technicians.
- CMC and GMP compliant documentation and tech transfer to CDMO.
- Servant Leader, team player – graduate of Leadership Academy at St Jude Children’s Research Hospital.
- Successful technology transfer of cGMP unit operations to Fujifilm Diosynth Biotechnologies, WuXi Biologics and Brammer Bio for rAAV downstream from harvest to fill/finish
Servant Leader, Director, Downstream Process Development, Applied Genetics Technologies Corporation [AGTC], Gainesville, Fl (July 2021 till Date)
- Successful adeno associated virus generation for tox/bio studies.
- Writing batch records compliant with Title 21 of CFR, Q5A for viral clearance; CMC guidance.
- Team building and leadership building – Hire and manage Scientists and their activities.
- Culture building – fostering a collaborative data driven environment.
- Hire and manage a team of Senior Scientists and plan their activities including training and orientation.
- Upstream and Downstream process development for manufacture of CRISPR-Cas9.
- Lentivirus - downstream unit operations from harvest to fill/finish
- cGMP manufacturing of antibody for neuroblastoma
- Upstream process development using Wave bioreactor / 200L Sartorius Cultibag and Downstream process development using AKTA ready and Alpha Plus TFF for monoclonal antibody production for treatment of neuroblastoma resulting in a US patent.
- Downstream cGMP manufacturing of Lentiviral vector for cell therapy with AKTA Ready
- Protein / Virus like particle expression in Baculovirus expression systems including BACMAM, HEK 293T cells, large scale E. coli expression, Protein / Virus like particle purification using different chromatographic techniques and AKTA FPLC systems.
- Baculovirus expression systems for protein / Virus like particle expression including BACMAM.
- Large scale E. coli expression
- Large-scale protein production and purification
- Bioprocess development.
- Lab management.
- Consultation with Principal Investigators
- Setup of pilot fermenter plant
Senior Research Technologist (2004 – 2006)
Protein Production Facility, Department of Infectious Diseases,
St. Jude Children Research Hospital, Memphis, TN, USA
- Baculovirus expression systems for protein expression
- Large scale E. coli expression
- Large-scale protein production and purification
- Set up of fermentation lab
2003 – 2004
Post-Doctoral Associate at the University of Tennessee, Vascular Biology Center of Excellence, Memphis, TN, USA
- Mapping of membrane associated proteins by phage display technology
- siRNA techniques for membrane associated protein
2002 – 2003
Research Scientist, Randox Laboratories, Northern Ireland, United Kingdom
- Production of recombinant proteins as disease markers
- Design of fermentation labs
1999 – 1998
DAAD Research fellowship
Protein researcher, (Cell immobilization group) Department of
Bioprocess Engineering (Technical University of Berlin,
Germany) (Part of my M. S. Study).
Skills:
Upstream operations
Molecular biology
Selection of expression vectors for viral vectors and proteins.
Development of cloning plan for viral vectors / recombinant proteins
Viral vector / protein expression using commercially available cell lines
Cell culture techniques
Isolation, development, propagation and maintenance of cell lines including insect cell, mammalian cells, E. coli and yeast.
Cell encapsulation, applications of encapsulated cell lines, development of encapsulation techniques.
Development of specific technologies for encapsulated hybridoma, insect and liver cell lines.
Cell migration assay, cell invasion assay, cell sorting, immuno-
fluorescence and FACS analysis.
Fermentation techniques
Development of scale-up process for production of viral vectors / proteins, from shake flask cultures to 200 l scale fermenters for E. coli, Yeast, Insect and mammalian cells.
Bioreactors handled include STR, Airlift, and Wave bioreactors including instruments from Braun Biotech, Sartorius, Wave Biotech, Applikon and New Brunswick Scientific; metabolite online analysis by Flownamics Seg-Flow and Nova Flex2
Downstream operations
Continuous flow centrifuges CEPA Z series, ultra-centrifugation Beckman and Sorvall series,
Depth Filtration with Sartorius PB1 filters, D0SP from Millipore
Cell lysis by Microfluidics Microfluidizer and GEA Ariete Homogenizer
Tangential flow filtration using Sartorius Alpha Plus, SciLog Millipore Pelicon, diafiltration, ultrafiltration for processing secreted proteins using instruments from Millipore.
Downstream Instrumentation skills
AKTA Ready, AKTA Avant, AKTA explorer 10S and 100, AKTA FPLC, AKTA Avant 25L and AKTA purifiers, Bio- RAD NGC platform including maintenance for protein purification.
Chromatographic techniques including Affinity, Ion exchange, Size exclusion, hydrophobic interaction, Tandem affinity and Multi-modal chromatography including prepacked and custom-made columns.
HPLC: Thermo Spray Inc. with RI detector using reverse phase and chiral columns
GC: Perkin Elmer gas chromatograph
Mass spectrometer: MALDI-TOF Finnigan electron spray ionization mass spectrometer coupled with HPLC and its analysis
IR spectrometer: Perkin Elmer infra-red spectrometer and peak detection
UV/Visible spectrometer: Pharmacia biotech and Kontron GMBH and Nanodrop.
Computer skills and Software
Latest version of Unicorn for AKTA including programming. MFCS for fermentation
Vector NTI, Genome database search tools including all BLAST tools, EMBL tools, CDART, Genome and Proteome FASTA, CLUSTALL, TRANSEQ, e-F sites, Vector NTI, Topology prediction, Unigene, SMART etc.
Languages C, C++ (Numerical recipes in C), G language (Virtual Instruments), MRU controls and Proportional Integral Controls (PID in Fermenters).
- Known software, Microsoft Word, Excel, Microsoft Access, Virtual instruments (National instruments), Design expert, Design ease, Stat ease, SPSS and Sigma plot.
Education:
1992–1996
The T.N. Dr. MGR Medical University – Chennai, India
Bachelor of Pharmacy
1997 –1999
Indian Institute of Technology - Madras, India/ Technical University of Berlin. Germany
Master of Science by Research DAAD fellowship
Research area: Cell biology, artificial organs by encapsulation
1999 – 2002
University of Ulster, Ireland, U.K.
Doctor of Philosophy (DPhil by Research)
Research area: Pharmaceutical biotechnology – Enzyme production and lipase catalyzed stereochemical biotransformation
Select Publications
- Oral Administration of Astrovirus Capsid Protein Is Sufficient To Induce Acute Diarrhea In Vivo Victoria A. Meliopoulos, Shauna A. Marvin, Pamela Freiden, Lindsey A. Moser, Prashant Nighot, Rizwana Ali, Anthony Blikslager, Muralidhar Reddivari, Richard J. Heath, Matthew D. Koci, Stacey Schultz-Cherry, doi: 10.1128/mBio.01494-16 1 November 2016 mBio vol. 7 no. 6 e01494-16
- Matthew M Wielgosz, Yoon-Sang Kim, Gael G Carney, Jun Zhan, Muralidhar Reddivari, Terry Coop, Richard J Heath, Scott A Brown, Arthur W Nienhuis. Generation of a lentiviral vector producer cell clone for human Wiskott-Aldrich syndrome gene therapy. Molecular Therapy — Methods & Clinical Development 2, 14063 (2015)
- Rebecca M Dubois, Pamela Freiden, Shauna Marvin, Muralidhar Reddivari, Richard J Heath, Stephen W White, Stacey Schultz-Cherry. Crystal structure of the avian astrovirus capsid spike. Journal of Virology 05/2013; DOI:10.1128/JVI.03139-12
- Rebecca M. DuBois, Hassan Zaraket, Muralidhar Reddivari, Richard J. Heath, Stephen W. White, Charles J. Russell. Acid Stability of the Hemagglutinin Protein Regulates H5N1 Influenza Virus Pathogenicity PLOS Pathogens, December 2011 | Volume 7 | Issue 12 | e1002398
Title: PROCESS FOR PRODUCING HU14.18K322A MONOCLONAL ANTIBODY; Document Type and Number: WIPO Patent Application WO/2019/161167, Kind Code: A1 2019
Nationality: American
E-mail: murali59_60002@yahoo.com
Education
Reddivari Muralidhar, PhD
570 Winding Valley Cove, Collierville, TN, 38017, USA
Murali59_60002@yahoo.com
9013783224
Experienced manager in therapeutic protein and virus expression and purification, bioprocess development including management of upstream and downstream process development and unit operations.
Professional Summary
- Over 20 years of experience in adeno associated virus / lentivirus / recombinant protein expression and purification using mammalian cells, insect cells, yeast and bacteria.
- Upstream Process development - Maintain and run SS Bioreactors / Fermentors from 10L to 200L, SUB wave-bioreactors up to 200L.
- Harvesting techniques including depth filtration, TFF and smart flow filtration.
- Downstream Process development - Maintain and run protein purification platforms including AKTA Ready, AKTA AVANT, AKTA Pure and AKTA purifiers. Bio-Rad NGC purification platforms: development of purification strategies based on protein chemistry.
- Fill/finish operations using ChaseLogeman / Bausch and Stroebel and SKAN isolators
- Manage and train a team of scientists / technicians.
- CMC and GMP compliant documentation and tech transfer to CDMO.
- Servant Leader, team player – graduate of Leadership Academy at St Jude Children’s Research Hospital.
- Successful technology transfer of cGMP unit operations to Fujifilm Diosynth Biotechnologies, WuXi Biologics and Brammer Bio for rAAV downstream from harvest to fill/finish
Servant Leader, Director, Downstream Process Development, Applied Genetics Technologies Corporation [AGTC], Gainesville, Fl (July 2021 till Date)
- Successful adeno associated virus generation for tox/bio studies.
- Writing batch records compliant with Title 21 of CFR, Q5A for viral clearance; CMC guidance.
- Team building and leadership building – Hire and manage Scientists and their activities.
- Culture building – fostering a collaborative data driven environment.
- Hire and manage a team of Senior Scientists and plan their activities including training and orientation.
- Upstream and Downstream process development for manufacture of CRISPR-Cas9.
- Lentivirus - downstream unit operations from harvest to fill/finish
- cGMP manufacturing of antibody for neuroblastoma
- Upstream process development using Wave bioreactor / 200L Sartorius Cultibag and Downstream process development using AKTA ready and Alpha Plus TFF for monoclonal antibody production for treatment of neuroblastoma resulting in a US patent.
- Downstream cGMP manufacturing of Lentiviral vector for cell therapy with AKTA Ready
- Protein / Virus like particle expression in Baculovirus expression systems including BACMAM, HEK 293T cells, large scale E. coli expression, Protein / Virus like particle purification using different chromatographic techniques and AKTA FPLC systems.
- Baculovirus expression systems for protein / Virus like particle expression including BACMAM.
- Large scale E. coli expression
- Large-scale protein production and purification
- Bioprocess development.
- Lab management.
- Consultation with Principal Investigators
- Setup of pilot fermenter plant
Senior Research Technologist (2004 – 2006)
Protein Production Facility, Department of Infectious Diseases,
St. Jude Children Research Hospital, Memphis, TN, USA
- Baculovirus expression systems for protein expression
- Large scale E. coli expression
- Large-scale protein production and purification
- Set up of fermentation lab
2003 – 2004
Post-Doctoral Associate at the University of Tennessee, Vascular Biology Center of Excellence, Memphis, TN, USA
- Mapping of membrane associated proteins by phage display technology
- siRNA techniques for membrane associated protein
2002 – 2003
Research Scientist, Randox Laboratories, Northern Ireland, United Kingdom
- Production of recombinant proteins as disease markers
- Design of fermentation labs
1999 – 1998
DAAD Research fellowship
Protein researcher, (Cell immobilization group) Department of
Bioprocess Engineering (Technical University of Berlin,
Germany) (Part of my M. S. Study).
Skills:
Upstream operations
Molecular biology
Selection of expression vectors for viral vectors and proteins.
Development of cloning plan for viral vectors / recombinant proteins
Viral vector / protein expression using commercially available cell lines
Cell culture techniques
Isolation, development, propagation and maintenance of cell lines including insect cell, mammalian cells, E. coli and yeast.
Cell encapsulation, applications of encapsulated cell lines, development of encapsulation techniques.
Development of specific technologies for encapsulated hybridoma, insect and liver cell lines.
Cell migration assay, cell invasion assay, cell sorting, immuno-
fluorescence and FACS analysis.
Fermentation techniques
Development of scale-up process for production of viral vectors / proteins, from shake flask cultures to 200 l scale fermenters for E. coli, Yeast, Insect and mammalian cells.
Bioreactors handled include STR, Airlift, and Wave bioreactors including instruments from Braun Biotech, Sartorius, Wave Biotech, Applikon and New Brunswick Scientific; metabolite online analysis by Flownamics Seg-Flow and Nova Flex2
Downstream operations
Continuous flow centrifuges CEPA Z series, ultra-centrifugation Beckman and Sorvall series,
Depth Filtration with Sartorius PB1 filters, D0SP from Millipore
Cell lysis by Microfluidics Microfluidizer and GEA Ariete Homogenizer
Tangential flow filtration using Sartorius Alpha Plus, SciLog Millipore Pelicon, diafiltration, ultrafiltration for processing secreted proteins using instruments from Millipore.
Downstream Instrumentation skills
AKTA Ready, AKTA Avant, AKTA explorer 10S and 100, AKTA FPLC, AKTA Avant 25L and AKTA purifiers, Bio- RAD NGC platform including maintenance for protein purification.
Chromatographic techniques including Affinity, Ion exchange, Size exclusion, hydrophobic interaction, Tandem affinity and Multi-modal chromatography including prepacked and custom-made columns.
HPLC: Thermo Spray Inc. with RI detector using reverse phase and chiral columns
GC: Perkin Elmer gas chromatograph
Mass spectrometer: MALDI-TOF Finnigan electron spray ionization mass spectrometer coupled with HPLC and its analysis
IR spectrometer: Perkin Elmer infra-red spectrometer and peak detection
UV/Visible spectrometer: Pharmacia biotech and Kontron GMBH and Nanodrop.
Computer skills and Software
Latest version of Unicorn for AKTA including programming. MFCS for fermentation
Vector NTI, Genome database search tools including all BLAST tools, EMBL tools, CDART, Genome and Proteome FASTA, CLUSTALL, TRANSEQ, e-F sites, Vector NTI, Topology prediction, Unigene, SMART etc.
Languages C, C++ (Numerical recipes in C), G language (Virtual Instruments), MRU controls and Proportional Integral Controls (PID in Fermenters).
- Known software, Microsoft Word, Excel, Microsoft Access, Virtual instruments (National instruments), Design expert, Design ease, Stat ease, SPSS and Sigma plot.
Education:
1992–1996
The T.N. Dr. MGR Medical University – Chennai, India
Bachelor of Pharmacy
1997 –1999
Indian Institute of Technology - Madras, India/ Technical University of Berlin. Germany
Master of Science by Research DAAD fellowship
Research area: Cell biology, artificial organs by encapsulation
1999 – 2002
University of Ulster, Ireland, U.K.
Doctor of Philosophy (DPhil by Research)
Research area: Pharmaceutical biotechnology – Enzyme production and lipase catalyzed stereochemical biotransformation
Select Publications
- Oral Administration of Astrovirus Capsid Protein Is Sufficient To Induce Acute Diarrhea In Vivo Victoria A. Meliopoulos, Shauna A. Marvin, Pamela Freiden, Lindsey A. Moser, Prashant Nighot, Rizwana Ali, Anthony Blikslager, Muralidhar Reddivari, Richard J. Heath, Matthew D. Koci, Stacey Schultz-Cherry, doi: 10.1128/mBio.01494-16 1 November 2016 mBio vol. 7 no. 6 e01494-16
- Matthew M Wielgosz, Yoon-Sang Kim, Gael G Carney, Jun Zhan, Muralidhar Reddivari, Terry Coop, Richard J Heath, Scott A Brown, Arthur W Nienhuis. Generation of a lentiviral vector producer cell clone for human Wiskott-Aldrich syndrome gene therapy. Molecular Therapy — Methods & Clinical Development 2, 14063 (2015)
- Rebecca M Dubois, Pamela Freiden, Shauna Marvin, Muralidhar Reddivari, Richard J Heath, Stephen W White, Stacey Schultz-Cherry. Crystal structure of the avian astrovirus capsid spike. Journal of Virology 05/2013; DOI:10.1128/JVI.03139-12
- Rebecca M. DuBois, Hassan Zaraket, Muralidhar Reddivari, Richard J. Heath, Stephen W. White, Charles J. Russell. Acid Stability of the Hemagglutinin Protein Regulates H5N1 Influenza Virus Pathogenicity PLOS Pathogens, December 2011 | Volume 7 | Issue 12 | e1002398
Title: PROCESS FOR PRODUCING HU14.18K322A MONOCLONAL ANTIBODY; Document Type and Number: WIPO Patent Application WO/2019/161167, Kind Code: A1 2019
Nationality: American
E-mail: murali59_60002@yahoo.com