Pilot scale production of a human monoclonal antibody against human immunodeficiency virus HIV-1.
Human monoclonal antibodies against the transmembrane protein gp41 of HIV-1 were isolated and purified on a pilot scale. A purification scheme was established for the production of human monoclonal antibodies on the gram scale. 50 1 of culture supernatant can be treated in one purification cycle. The hybridomas were mass cultured in an airlift fermenter. The culture broth was clarified by microfiltration and chromatographed on CM-Sepharose fast flow and protein A Superose. Scale up of the high performance affinity chromatography from 1 ml protein A Superose up to 40 ml is described. All desalting steps were performed by gel filtration on Sephadex G-25 coarse. The yield of the whole purification procedure is in the range of 50-60%. The purity is higher than 99.9%. DNA and reverse transcriptase could not be detected. The whole method is designed as a basis for scale up to industrial scale. Results from quality control assays have proven the validity of this approach.
Jungbauer
A
A
Institute of Applied Microbiology, University of Agriculture and Forestry, Vienna, Austria.
Tauer
C
C
Wenisch
E
E
Steindl
F
F
Purtscher
M
M
Reiter
M
M
Unterluggauer
F
F
Buchacher
A
A
Uhl
K
K
Katinger
H
H
D000911:Antibodies, Monoclonal / Q000302:isolation & purification*; D015153:Blotting, Western; D002846:Chromatography, Affinity / Q000379:methods; D002850:Chromatography, Gel / Q000379:methods; D002852:Chromatography, Ion Exchange / Q000379:methods; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D015483:HIV Antibodies / Q000302:isolation & purification*; D015497:HIV-1 / Q000276:immunology*; D006801:Humans; D006825:Hybridomas / Q000276:immunology; D007074:Immunoglobulin G / Q000032:analysis / Q000302:isolation & purification*
D000911:Antibodies, Monoclonal; D015483:HIV Antibodies; D007074:Immunoglobulin G