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Production of Antibody Fragments in Escherichia coli
Author(s) -
HARRISON J. S.,
KESHAVARZMOORE E.
Publication year - 1996
Publication title -
annals of the new york academy of sciences
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.712
H-Index - 248
eISSN - 1749-6632
pISSN - 0077-8923
DOI - 10.1111/j.1749-6632.1996.tb40556.x
Subject(s) - citation , library science , metabolic engineering , computer science , chemistry , biochemistry , enzyme
As a result of the increasing commercial demand for antigen-binding antibody fragments for medical, industrial, and diagnostic applications, a need exists for a production process whereby these proteins may be produced efficiently and at low cost. Bacterial expression in, for example, Escherichia coli, where genetic manipulations and fermentation have been widely investigated, provides an alternative to animal cell culture. Large-scale processes for production of recombinant proteins from E. coli have been reported,’ even though in certain expression strategies where the recombinant protein is expressed in an insoluble form, bacterial expression may still be less economical.2 Reviews of recent literature concerned with antibody e n g i n e ~ r i n g ~ ~ and the expression of antibody fragments in E. coli have already been published.”8 Although antibody fragments have been expressed in an inactive form as inclusion bodies in the cytoplasm, a more common strategy is the direction of antibody fragments to the periplasmic space where folding to active antigen-binding proteins may occur. Very few published reports to date have been concerned specifically with aspects of the fermentation of E. coli to produce high levels of functional antibody fragments. Nevertheless, it is clear that the expression of functional antibody fragments is affected by the nature of the expression-inducer system as well as the fermentation parameters. Much of this work has been performed in shake flasks where production levels have been low, but reports of concentrations of up to 450 mg/l in fermentcr cultures of Fv fragmentsY and 1 to 2 g/l of bivalent antibody fragmentsi0 have been made. This paper reviews recent literature on the production of antibody fragments by fermentation of E. coli with a view to integration with subsequent downstream processing. With this in mind, it is useful to relate the production of antibody fragments to more general information concerning recombinant protein production in E. coli.