QuantyBio Corporation is developing a monoclonal antibody and they need your help. They need to make preclinical (PK and toxicology) and Clinical (Phase I) material for their clinical program. The company has developed a cell line that produces about 600 mg/L in a batch culture in 7 days. Cell density: Start at 0.5 MM/mL, max 15 MM/mL, end viability = 40%, Specific productivity of these cells is at qp = 35 pg/cell/day. The cells have a specific oxygen consumption of qO2 = 0.22 uM/MM-hr, a research cell bank (RCB) is made and kept frozen in LN2 freezer (a total of 20 vials of RCB).
They would like to develop a fed-batch process and optimize it. The targeted process will look like this:
Starting cell density: 0.5 Million cells/mL
Titer = 2 g/L
Culture duration = 14 days
Viability at the end of the culture > 50%
Chemically defined medium and feeds
3-step purification step
Overall yield > 50% (bioreactor to final vial)
Final formulation: 25 mg/mL
Fill volume: 2 ml in a 3ml vial (liquid formulation)
The size of the preclinical and clinical trials are as follows:
Preclinical: 300 vials are needed from a GLP lab
Clinical: 5000 vials are needed from a GMP facility
1. Preclinical Material:
Calculate the amount of material (in grams) to be made in the bioreactor based on preclinical trial needs
Calculate the size of the bioreactor (in Liters) to be used
Based on answer (1b) above:
1. Calculate the dimensions of the bioreactor, Diameter, Height, Impeller diameter, Cross-sectional area (A), etc. In terms of the impeller, the bioreactor for the preclinical study will have a pitched blade impeller operated at 200 rpm.
2. What are the Oxygen Transfer Rate (OTR) Requirements of the cells at a maximum cell density of 20 MM/mL?
3. What is the kla requirement if pure O2 is used in the gas and the DO is maintained at 50% air saturation?
4. The bioreactor for the preclinical study will have a microsparger and it can maintain DO at 20 MM/mL densities with a gas flow rate of 1 Liter per minute. What is the vvm and what is the gas superficial velocity (u=Qg/A)?
5. Using kla = a(RPM)^b(vvm)^c correlation with b = 0.35 and c = 0.75, what is the value of a for kla correlation?
2. Clinical material:
Calculate the amount of material (in grams) to be made in the bioreactor based on Phase I trial needs
Calculate the size of the bioreactor (in Liters) to be used
Based on answer (2b) above:
6. Calculate the dimensions of the bioreactor, Diameter, Height, Impeller diameter, Cross-sectional area (A), etc.
7. What should be the agitation rate for the GMP bioreactor (the small bioreactor uses 200 rpm)?
8. Using the kla correlation kla = a(RPM)^b(vvm)^c with b = 0.35 and c = 0.75. What is the vvm of oxygen needed in the GMP bioreactor?
9. What is the gas superficial velocity (u=Qg/A) in the GMP bioreactor?
Part B: Bioreactor Design
Using the formulas presented in this module for a chemostat, calculate and plot as a function of dilution rate (between 0.2-2 1/day, with an increment of 0.05), cell density, and substrate concentration using the following parameters (2 points): um = 1.8, Ks = 0.3, Yx/s = 1, So = 1
Using the formulas presented in this module for perfusion, calculate and plot as a function of dilution rate (between 0.2-4 1/day, with an increment of 0.05), cell density, and substrate concentration using the following parameters: um = 1.8, Ks = 0.3, Yx/s = 1, So = 1, alpha = 0.1, C = 2