Monoclonal antibody production with on-line harvesting and process monitoring.
A semi-automated system has been designed for on-line harvesting and monitoring of monoclonal antibody (mAb) production. [The antibody was directed against the peptide AGPAGTGKTTKDL.] Analytical and purification units were interfaced to the fermenter via a hollow fiber cartridge in which fermentation broth was continuously circulated through the lumen of the hollow fiber system. Permeate from the hollow fiber cartridge was pumped through either an analytical sampling loop or a preparative Protein G column where antibody species were captured. Switching between monitoring and harvesting was achieved by two 3-way toggle valves. Samples from the analytical sampling loop were transported to an analytical Protein G chromatography column for quantitation of all immunoglobulin G species in the fermenter. Data acquisition and processing was performed by the data system of the liquid chromatograph. All valves in the system except the two toggle valves were controlled by the liquid chromatograph. Antibody biosynthesis was monitored for the first 60 h of fermentation. Harvesting was initiated when mAb accumulated in the fermenter. Complete harvesting took approximately 90 h.
Zhang
J
J
Department of Chemistry, Purdue University, W. Lafayette, IN 47903, USA.
Zhou
H
H
Ji
Z
Z
Regnier
F
F
D000595:Amino Acid Sequence; D000911:Antibodies, Monoclonal / Q000032:analysis / Q000096:biosynthesis* / Q000302:isolation & purification; D002021:Buffers; D002853:Chromatography, Liquid; D005285:Fermentation; D006825:Hybridomas / Q000378:metabolism; D007074:Immunoglobulin G / Q000737:chemistry / Q000302:isolation & purification; D007202:Indicators and Reagents; D008969:Molecular Sequence Data; D009419:Nerve Tissue Proteins / Q000737:chemistry; D009862:Online Systems; D010455:Peptides / Q000276:immunology; D011494:Protein Kinases / Q000737:chemistry
D000911:Antibodies, Monoclonal; D002021:Buffers; C029152:G-substrate; D007074:Immunoglobulin G; D007202:Indicators and Reagents; D009419:Nerve Tissue Proteins; D010455:Peptides; D011494:Protein Kinases