Document 0174
 DOCN  M9460174
 TI    Stable, continuous large-scale production of human monoclonal HIV-1
       antibody using a computer-controlled pilot plant.
 DT    9404
 AU    Unterluggauer F; Doblhoff-Dier O; Tauer C; Jungbauer A; Gaida T; Reiter
       M; Schmatz C; Zach N; Katinger H; Institute of Applied Microbiology,
       University of Agriculture and; Forestry, Vienna, Austria.
 SO    Biotechniques. 1994 Jan;16(1):140-7. Unique Identifier : AIDSLINE
       MED/94183545
 AB    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)
 DE    Antibodies, Monoclonal/*BIOSYNTHESIS/ISOLATION & PURIF  Cell Line
       Computers  Human  HIV Antibodies/*BIOSYNTHESIS/ISOLATION & PURIF
       HIV-1/*IMMUNOLOGY  JOURNAL ARTICLE

       SOURCE: National Library of Medicine.  NOTICE: This material may be
       protected by Copyright Law (Title 17, U.S.Code).