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Production of a crucial therapeutic protein for Trauma and Accident care in Escherichia Coli system

Primary Information

Domain

Healthcare

Project No.

4192

Sanction and Project Initiation

Sanction No: F.NO. 41-2/2015 -T-S-I (PT.) SL. NO. 4192

Sanction Date: 1/9/2017

Project Initiation date: 30/08/2017

Project Duration: 36

Partner Ministry/Agency/Industry

Indian Institute of Technology Delhi

 

Role of partner:Infrastructural support Financial Support

 

Support from partner:Infrastructural support Financial Support

Principal Investigator

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Tapan Kumar Chaudhuri
Indian Institute of Technology -Delhi

Host Institute

Co-PIs

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Prof. K. J. Mukherjee
Jawahar Lal Nehru University, Delhi

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Prof. Amit Kumar Dinda
All Indian Institute of Medical Science

 

Scope and Objectives

Development of an efficient E. coli based system for enhanced production of recombinant Human Serum Albumin protein. Optimization of expression, folding, purification and conditions for improving shelf life of the E. coli derived rHSA. Characterization by carrying out structural, physiological, biochemical, functional studies of the E. coli derived rHSA and its comparison with the plasma derived form. Development of the bio-processing method for scaling-up of the production of rHSA derived from E. coli. To carry out Toxicological studies for safety evaluation of rHSA for clinical usage.

Deliverables

Promoting the very well-studied E .coli host system, which is of low cost, and scalable, to prepare immensely important, multi- application oriented and immensely demand driven recombinant therapeutic protein HSA. Development of comprehensive strategy for obtaining higher yield of functional rHSA through improvement of cell growth condition for enhancement of protein expression (Bioprocess development), modulating in vivo folding of nascent recombinant protein (cellular folding), and optimization of the process of extraction and purification (downstream processing). To deliver good amount of contamination free recombinant HSA, having a reasonable shelf life, for use in trauma and accidental care in an affordable price.

 

Scientific Output

Currently, HSA for treatment of trauma and accident is derived from human blood which is the major issue of concern as it possesses the risk of contamination by various harmful blood-derived pathogenic virus like HIV, Hepatitis etc. and also is in limited supply. Whereas, development of a strategy for obtaining an optimized E. coli based system for enhanced production of functionally active soluble rHSA with improved shelf life is the need of the hour and has potential to overcome the above mentioned risk by production of stable, pathogen free rHSA crucial therapeutic biomolecule of trauma care for clinical usage in sufficient quantities in a cost effective manner.

 

Results and outcome till date

Currently, HSA for treatment of trauma and accident is derived from human blood which is the major issue of concern as it possesses the risk of contamination by a various harmful blood-derived pathogenic virus like HIV, Hepatitis, etc. and also is in limited supply. Whereas, development of a strategy for obtaining an optimized E. coli based system for enhanced production of functionally active soluble rHSA with an improved shelf life is the need of the hour and has the potential to overcome the above-mentioned risk by the production of stable, pathogen-free rHSA crucial therapeutic biomolecule of trauma care for clinical usage in sufficient quantities in a cost-effective manner.

 

Societal benefit and impact anticipated

Patients undergoing trauma and accidental care in various places will be benefitted; since HSA has immense potential and wide applications in the biotechnology sector also, therefore the biotechnological industry will also get a benefit with the development of the product i.e. E. coli derived rHSA. For now, the experiments will be carried out to check the toxicological status of the E. coli derived rHSA for its clinical usage, which will be carried out using animal models (rats) & peripheral blood from human volunteers.

Next steps

An optimized Downstream processing protocol for purification of functional rHSA protein. Biophysical characterization, stability, and comparison of E.coli derived rHSA with the commercially available product. Developing Bioprocess strategy for scaling up production of rHSA in E. coli. Formulation development for Pre-clinical trials.

Scholars and Project Staff

Project Staff appointment date: 01 Sept 2017 - 08 Nov 2017.
Junior Research Fellow appointment date: 10 Nov 2017 - 09 Nov 2019

Challenges faced

Difficulty with enhancing the solubility of the rHSA when expressed in E. coli Difficulty in achieving required purity levels.

Financial Information

  • Total sanction: Rs. 6000000

  • Amount received: Rs. 4840000

  • Amount utilised for Equipment: Rs. 2291927

  • Amount utilised for Manpower: Rs. 583850

  • Amount utilised for Consumables: Rs. 1771743

  • Amount utilised for Contingency: Rs. 61740

  • Amount utilised for Travel: Rs. 0

  • Amount utilised for Other Expenses: 0

  • Amount utilised for Overheads: Rs. 0

Equipment and facilities

 

AKTA pure Thermo Scientific Refrigerator