Control of cell growth through a temperature feedback loop

Abstract

The present invention relates to the use of online biomass capacitance monitoring in cultures as a way to control the growth of cells through the use of a temperature control loop. In certain embodiments, a biomass capacitance probe is used to measure the cell density, and a predetermined growth curve is used to adjust the temperature in the culture.

1 . A method of controlling the growth of cells in a bioreactor comprising:
 a) measuring the viable cell density of the cells growing in culture using a biomass capacitance probe; and   b) utilizing a temperature control loop to adjust the growth rate of the cells.   
     
     
         2 . A method of modulating reactor temperature to control the growth of cells in a bioreactor comprising:
 a) measuring the viable cell density of the cells growing in culture using a biomass capacitance probe; and   b) utilizing a temperature control loop to modulate the reactor temperature and adjust the growth rate of the cells.   
     
     
         3 . A method of minimizing the growth fluctuation of cells in a bioreactor comprising:
 a) measuring the viable cell density of cells growing in culture using a biomass capacitance probe; and   b) utilizing a temperature control loop to adjust the growth rate of the cells.   
     
     
         4 . The method of  claim 1 , wherein a pre-determined growth curve is utilized to adjust the temperature in the culture. 
     
     
         5 . The method of  claim 1 , wherein a bolus feed is added to the bioreactor daily. 
     
     
         6 . The method of  claim 5 , wherein the daily bolus feed is begun starting on about day 1, about day 1.5, about day 2, about day 2.5, about day 3, about day 3.5, about day 4, about day 5, about day 6, about day 7, about day 8, about day 9 or about day 10 after the culture is inoculated. 
     
     
         7 . The method of  claim 5 , wherein the daily bolus feed volume is about 1%, about 2%, about 3%, about 4%, about 5%, about 6%, about 7%, about 8%, about 9%, about 10%, about 15% or more of the initial working volume. 
     
     
         8 . The method of  claim 5 , wherein the daily bolus feed volume is about 3%, about 3.1%, about 3.2%, about 3.3%, about 3.4%, about 3.5%, about 3.6%, about 3.7%, about 3.8, about 3.9% or about 4% of the initial working volume. 
     
     
         9 . The method of  claim 5 , wherein the daily bolus feed volume is about 3.6% of the initial working volume. 
     
     
         10 . The method of  claim 1 , wherein a bolus feed is added to the bioreactor hourly. 
     
     
         11 . The method of  claim 10 , wherein the hourly bolus feed is begun starting on about day 1, about day 1.5, about day 2, about day 2.5, about day 3, about day 3.5, about day 4, about day 5, about day 6, about day 7, about day 8, about day 9 or about day 10 after the culture is inoculated. 
     
     
         12 . The method of  claim 10 , wherein the hourly bolus feed is about 1/24 of the calculated volume for a daily bolus feed. 
     
     
         13 . The method of  claim 10 , wherein the hourly bolus feed is about 0.1%, about 0.11%, about 0.12%, about 0.13%, about 0.14%, about 0.15%, about 0.16%, about 0.17%, about 0.18%, about 0.19%, about 0.2%, about 0.25%, about 0.3%, about 0.4% or about 0.5% of the initial working volume. 
     
     
         14 . The method of  claim 10 , wherein the hourly bolus feed volume is about 0.15% of the initial working volume. 
     
     
         15 . The method of  claim 1 , wherein the viable cell density is plotted to a temperature range of about 30-40° C. 
     
     
         16 . The method of  claim 1 , wherein the viable cell density is plotted to a temperature range of about 31-37° C. 
     
     
         17 . The method of  claim 1 , wherein daily temperature oscillations in the reactor have a peak to peak amplitude of about 10° C., about 9° C., about 8° C., about 7° C., about 6° C., about 5° C., about 4° C., about 3° C., about 2° C., about 1° C. or less. 
     
     
         18 . The method of  claim 1 , wherein the cells produce a polypeptide of interest. 
     
     
         19 . The method of  claim 18 , wherein the polypeptide of interest is an antibody. 
     
     
         20 . The method of  claim 1 , wherein the cells are mammalian cells. 
     
     
         21 . The method of  claim 1 , wherein the cells are Chinese Hamster Ovary (CHO) cells. 
     
     
         22 . The method of  claim 1 , wherein the biomass capacitance probe is an INCYTE probe. 
     
     
         23 . The method of  claim 1 , wherein the pH of the culture is about 6.5 to about 8.0, about 7.0 to about 8.0, or about 7.0 to about 7.5. 
     
     
         24 . The method of  claim 1 , wherein the pH of the culture is about 7.1 to about 7.4. 
     
     
         25 . The method of  claim 1 , wherein the culture is inoculated with cells at a density of about 1×10 5 , about 2×10 5 , about 3×10 5 , about 4×10 5 , about 5×10 5 , about 6×10 5 , about 7×10 5 , about 8×10 5 , about 9×10 5 , or about 10×10 5  cells/ml. 
     
     
         26 . The method of  claim 1 , wherein the culture is inoculated with cells at a density of about 6×10 5  cells/ml.