Bids Are Invited For Custom Bid for Services - Development contract for IL6 Production - Production of Recombinant IL-6 Process development approach for the production of recombinant IL6 (rIL6): The process development of rIL6 should be performed implementing the guidelines of International Conference on Harmonisation (ICH) to develop the process and in process product controls for the commercial manufacture of rIL6. It should involve the following steps: 1. Construct development for recombinant Mouse and Human IL-6. It should involve following steps: a) In silico analysis, Codon biasing and Gene synthesis. b) Cloning in bacterial expression vector, eukaryotic and mammalian expression vector. c) Primary & Secondary verification of sequence. d) Screening for best expression colony. 2. Process establishment at shake flask level for final clone selection. It should involve following steps: a) Standardization of Physical parameters for expression optimization b) Downstream Process (DSP) optimization c) Analytical characterization Milestone 1: Construct synthesis, successful expression of clones. Small scale production of Mouse and Human recombinant IL-6 (100 ug each) which will be tested in vitro and in vivo for biomarker induction along with the complete certificate of analysis data. 3. Scale up the expression and process development at 5L scale of the best suited recombinant IL-6 and host. It should involve following steps: a) Process development at bioreactor level b) Multiple Parameters like High cell density batches, time, temperature, Induction OD, Inducer concentration etc. should be optimized for getting high expressing clone. c) Complete analytics assessment against commercially available rIL6 d) Method development and qualification e) Stability studies f) Dossier preparation 4. Consistency batches a) Three consistency batches for process reproducibility at manufacturing batch process scale utilizing draft batch production records. b) One batch – For Toxicology study. Milestone 2: Scale up the development of best suited recombinant IL-6 from mouse (10 mg) and human (100 mg total from eukaryotic and mammalian clone) at 5L scale. THE FOLLOWING STEPS SHOULD BE INVOLVED FOR PROCESS DEVELOPMENT: Cell banking: a) Clone that show expression at small scale and further tested for activity of rIL6 should be considered and stocked as Research cell bank (RCB). b) The selected clone should be utilized for further process development tested at 1L & 5L scale followed by bio-physical characterization and testing of activity profile. c) For final selected clone, cell banking system of Master Cell Bank (MCB) and Working Cell Bank (WCB) should be developed and maintained in accordance to ICH guidelines along with full description of the procedures followed. d) Characterisation including identity, viability, stability, presence of adventitious agents should be conducted as per the ICH guidelines. Upstream process development Fermentation conditions and in-process controls (IPC) should be optimised, developed, established and documented as batch manufacturing records. Process validation Process validation should include the collection and evaluation of data, from the process design stage throughout production, establishing scientific evidence that the optimized process is capable of consistently delivering a quality drug substance. Development, characterisation, and validation of the rIL6 production process should be established based on previous studies on recombinant IL6, understanding of similar molecules and processes using similar host platform and analysis of recombinant IL6 (rIL6) B. Biological characterisation: Functional activity of rIL6 Shall be accessed by a. Cell proliferation of mouse B9/TF1 cells should be analysed. b. Induction of biomarker at Source/ INMAS C. Variants and impurities Presence of any process-related impurities like Host cell proteins (HCP) and DNA, Fermentation process derived impurities, Downstream-derived impurities such as leachables during chromatography Other impurities including endotoxin, bioburden etc. should be analysed. The impurity profile of rIL6 active substance should be analysed for presence of fragments, dimers and higher aggregates D. Adventitious agents Usage of animal-derived raw materials is strictly prohibited in the rIL6 manufacturing process. Details of all raw material procured and defined for rIL6 production process should be documented. E. Product Manufacturing To develop and deliver the constructs, purified rIL6 and a manufacturing process to a scale of 5 litres to deliver rIL6 that should be used by INMAS-DRDO for its further in-vitro and in-vivo animal studies. The complete manufacturing process should start with making and maintenance of cell banks, and should describe the fermentation, purification, formulation steps, including IPC and final QC. F. Product specification Description of physical appearance of active substance, final formulation, solubility, storage conditions should be described and finalized as the Target product profile. Description of complete analytical method development procedure should be described and documented. The test material should be tested using these methodologies to match the required specifications, as follows. G. Stability The design of the stability program should be in accordance with the ICH guidelines. The stability data should detail the storage conditions, shelf life of rIL6 and other details relating to the final storage specifications of the final protein. Deliverables: 1. The constructs (mouse and human recombinant IL6 in bacterial, eukaryotic and mammalian expression system), 2. Purified proteins (10 mg mouse and 100 mg human rIL6) 3. A manufacturing process to a scale of 5 litres for production of active drug substance (rIL6) Eligibility criteria for the source firm/ Research Service Provider: a) Should have DSIR Recognition. b) Biosafety assessment status of the Research Service Provider should be declared. c) Should have specialization in cloning, fermentation and recombinant protein expression (in E.coli, Pichia & Mammalian hosts), refolding and purification with more than 5 years of research experience. d) The Research Service Provider should have experience of doing minimum 3 projects on similar platforms from Ideation to protein delivery stage for different therapeutic area including vaccine, Biosimilar, recombinant proteins, hormones etc. Provide project sanction letter and satisfactory delivery report. e) The RSP should have Strong background on protein expression in different hosts from easy to difficult to express proteins from plate level to a bioreactor level. f) Should have well established facility for the development and product generation. g) The RSP Should have established and well equipped labs and culture rooms for Microbial and Yeast Systems. h) Should have well established bioreactor facility for process development and pilot scale fermentation for microbial and yeast systems ranging from 3L to 300L bioreactor Total Quantity :
