TY - JOUR

T1 - Metabolism of hybridoma cells and antibody secretion at high cell densities in dialysis tubing

AU - Käsehagen, Claudia

AU - Linz, Fritjof

AU - Kretzmer, Gerlinde

AU - Scheper, Thomas

AU - Schügerl, Karl

N1 - Funding information: Part of these investigations were carried out in the frame of the BAP-project 0132 D. The authors gratefully acknowledge the financial support of the European Community.

PY - 1991/11

Y1 - 1991/11

N2 - The experimental setup, consisting of a bundle of dialysis tubing 2.5 mm in diameter [10-15 kD cutoff, mean pore size 25 Å, 20 μm (dry) and 40 μm (wet) wall thickness] inserted into a 1-l glass bioreactor supplied with oxygen and pH electrodes, a porous gas distributor, a sampling tube, and a holder for the eight pieces of dialysis tubing, was developed to investigate the properties and the microenvironment of hybridoma cells enclosed in the tubing during their batch cultivation. The concentrations of low-molecular-weight medium components were the same inside and outside the tubing, and it was possible to control the microenvironment of the cells in the tubing easily. The cell damage caused by mechanical stress was less in the dialysis tubing than in stirred spinner flasks. The influence of the initial cell density in the range from 4 × 105 to 1 × 108 cells ml-1 and the cultivation time were evaluated according to the total and viable cell concentrations and the cell/cell fragment size distributions. Furthermore, the cell membrane properties, glucose consumption rate, lactate, ammonia and lipid storage material, and the monoclonal antibody production rates as well as intracellular enzyme activities in the culture medium were measured and compared to those in reference cultures in spinner flasks with the same inoculum at low initial cell densities. In dialysis tubing in a concentration range of 5 × 106 to 108 cells ml-1, the total and viable concentrations of cells remained the same during cultivation. At high cell concentrations, cell size decreased, membrane folding increased, the fraction of dead cells and cell fragments increased, and the metabolic activity of the cells and the monoclonal antibody productivity were higher than with low cell concentrations. Since the dialysis membrane is nonpermeable for proteins, high monoclonal antibody concentrations were attained.

AB - The experimental setup, consisting of a bundle of dialysis tubing 2.5 mm in diameter [10-15 kD cutoff, mean pore size 25 Å, 20 μm (dry) and 40 μm (wet) wall thickness] inserted into a 1-l glass bioreactor supplied with oxygen and pH electrodes, a porous gas distributor, a sampling tube, and a holder for the eight pieces of dialysis tubing, was developed to investigate the properties and the microenvironment of hybridoma cells enclosed in the tubing during their batch cultivation. The concentrations of low-molecular-weight medium components were the same inside and outside the tubing, and it was possible to control the microenvironment of the cells in the tubing easily. The cell damage caused by mechanical stress was less in the dialysis tubing than in stirred spinner flasks. The influence of the initial cell density in the range from 4 × 105 to 1 × 108 cells ml-1 and the cultivation time were evaluated according to the total and viable cell concentrations and the cell/cell fragment size distributions. Furthermore, the cell membrane properties, glucose consumption rate, lactate, ammonia and lipid storage material, and the monoclonal antibody production rates as well as intracellular enzyme activities in the culture medium were measured and compared to those in reference cultures in spinner flasks with the same inoculum at low initial cell densities. In dialysis tubing in a concentration range of 5 × 106 to 108 cells ml-1, the total and viable concentrations of cells remained the same during cultivation. At high cell concentrations, cell size decreased, membrane folding increased, the fraction of dead cells and cell fragments increased, and the metabolic activity of the cells and the monoclonal antibody productivity were higher than with low cell concentrations. Since the dialysis membrane is nonpermeable for proteins, high monoclonal antibody concentrations were attained.

KW - electrorotation measurements

KW - flow cytometry

KW - high cell density

KW - Hybridoma cells

KW - membrane reactor

KW - monoclonal antibody

UR - http://www.scopus.com/inward/record.url?scp=0026261763&partnerID=8YFLogxK

U2 - 10.1016/0141-0229(91)90103-H

DO - 10.1016/0141-0229(91)90103-H

M3 - Article

C2 - 1367998

AN - SCOPUS:0026261763

VL - 13

SP - 873

EP - 881

JO - Enzyme and microbial technology

JF - Enzyme and microbial technology

SN - 0141-0229

IS - 11

ER -