Stable, continuous large-scale production of human monoclonal HIV-1 antibody using a computer-controlled pilot plant.
A completely automated pilot plant used for fermentation has been employed with direct digital control (DDC) technology for monitoring and regulating growth of human cells. A human hybridoma cell line (3D6) producing anti-human immunodeficiency virus (HIV)-1 antibodies was used as a model for large-scale production (300-liter airlift fermentor) in continuous culture. Parameters controlled were pH, dissolved oxygen, temperature and the flow rate of four gases used in the process. A control strategy was implemented to achieve constant fluid velocity and mixing by maintaining the rate of gas flow at a constant level. Another advantage of this approach was that the total gas flow required for optimal fluid circulation was reduced from 1 volume gas/volume fermenter/hour (vvh) to 0.3 vvh. Use of a low flow rate eliminated the serious problems of foaming, which contributed significantly to cell destruction, shorter filter-life and other considerations. Dilution rate was optimized at laboratory scale for maximum productivity, which results in relatively low viability. At a dilution rate of 0.0076 h-1, a total cell density of 6-7 x 10(5) cells/ml with a viability of approximately 75% was maintained during long-term continuous cultivation. These growth conditions resulted in a product titer stabilized in the range of 35 micrograms IgG/ml. Batchwise purification was achieved with a recovery of more than 50% and a final purification of active monoclonal antibody representing about 99% product. Results from isoelectric focusing and Western blotting demonstrated batch-to-batch consistency of the purified human monoclonal antibody to HIV-1 during the continuous growth process.(ABSTRACT TRUNCATED AT 250 WORDS)
Unterluggauer
F
F
Institute of Applied Microbiology, University of Agriculture and Forestry, Vienna, Austria.
Doblhoff-Dier
O
O
Tauer
C
C
Jungbauer
A
A
Gaida
T
T
Reiter
M
M
Schmatz
C
C
Zach
N
N
Katinger
H
H
D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000302:isolation & purification; D002460:Cell Line; D003201:Computers; D015483:HIV Antibodies / Q000096:biosynthesis* / Q000302:isolation & purification; D015497:HIV-1 / Q000276:immunology*; D006801:Humans
D000911:Antibodies, Monoclonal; D015483:HIV Antibodies