ONLINE MONITORING OF MONOCLONAL-ANTIBODY FORMATION IN HIGH-DENSITY PERFUSION CULTURE USING FIA

被引：31

作者：

FENGE, C

FRAUNE, E

FREITAG, R

SCHEPER, T

SCHUGERL, K

机构：

[1] B.Braun Diessel Biotech GmbH, Melsungen, D-3508

[2] Institut für Technische Chemie, Universität Hannover, Hannover, D-3000

来源：

CYTOTECHNOLOGY | 1991年 / 6卷 / 01期

关键词：

FLOW INJECTION ANALYSIS; HIGH CELL DENSITY; HYBRIDOMA CELLS; ONLINE MONITORING OF MONOCLONAL ANTIBODY FORMATION; ONLINE GLUCOSE AND LACTATE MEASUREMENTS; PERFUSION CULTIVATION;

D O I：

10.1007/BF00353703

中图分类号：

Q81 [生物工程学（生物技术）]; Q93 [微生物学];

学科分类号：

071005 ; 0836 ; 090102 ; 100705 ;

摘要：

An automated flow injection system for on-line analysis of proteins in real fermentation fluids was developed by combining the principles of stopped-flow, merging zones flow injection analysis (FIA) with antigen-antibody reactions. IgG in the sample reacted with its corresponding antibody (a-IgG) in the reagent solution. Formation of insoluble immunocomplexes resulted in an increase of the turbidity which was determined photometrically. This system was used to monitor monoclonal antibody production in high cell density perfusion culture of hybridoma cells. Perfusion was performed with a newly developed static filtration unit equipped with hydrophilic microporous tubular membranes. Different sampling devices were tested to obtain a cell-free sample stream for on-line product analysis of high molecular weight (e.g., monoclonal antibodies) and low molecular weight (e.g., glucose, lactate) medium components. In fermentation fluids a good correlation (coefficient: 0.996) between the FIA method and an ELISA test was demonstrated. In a high density perfusion cultivation process mAb formation was succesfully monitored on-line over a period of 400 h using a reliable sampling system. Glucose and lactate were measured over the same period of time using a commercially available automatic analyser based on immobilized enzyme technology.

引用

页码：55 / 63

页数：9

共 9 条

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