PMID: 9005945Jan 15, 1997Paper

High level production of soluble single chain antibodies in small-scale Escherichia coli cultures

Journal of Immunological Methods
S M KipriyanovM Little

Abstract

We have investigated the effect of growth and induction conditions on the production of soluble single-chain Fv antibody fragments in Escherichia coli under the control of wt lac promoter. The scFv was directed into the periplasmic space by a pelB leader sequence. Addition of sucrose to the medium gave a 15-25-fold increase in the yield of soluble scFv-phOx (3.0 mg/l) for bacterial shake-tube cultures and an increase of 80-150-fold (16.5 mg/l) for shake-flask cultures. Using flask culture in the presence of 0.4 M sucrose, a significant amount of scFv was released into the medium. We found that the scFv could be made to accumulate in the periplasm or be secreted into the medium by simply changing the incubation conditions and the concentration of the inducer. The ratio between soluble antibody fragments and insoluble scFv aggregates proved to be dependent on the strength of the promoter. Lowering the incubation temperature below 20 degrees C had no effect on the yield of soluble antibody fragments in the periplasm, but they were no longer secreted into the medium. An example of high level production in shake-flask cultures and one-step purification by immobilized metal affinity chromatography (IMAC) is described for a soluble sc...Continue Reading

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Citations

Feb 24, 2004·Folia Microbiologica·D Blazek, V Celer
Oct 27, 2012·Applied Microbiology and Biotechnology·Randy Chi Fai CheungTzi Bun Ng
May 30, 2008·European Journal of Nuclear Medicine and Molecular Imaging·Matthias EderMichael Eisenhut
Jan 13, 2006·Cancer Metastasis Reviews·Mehdi Arbabi-GhahroudiRoger MacKenzie
Jan 25, 2013·BioDrugs : Clinical Immunotherapeutics, Biopharmaceuticals and Gene Therapy·Mohammad R TohidkiaYadollah Omidi
Nov 1, 2002·Biochemical and Biophysical Research Communications·Kim Bak JensenPeter Kristensen
Aug 21, 2003·Clinica Chimica Acta; International Journal of Clinical Chemistry·Gordana WozniakFlorian Rüker
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Sep 15, 2000·Journal of Immunological Methods·B E Power, P J Hudson
Feb 27, 2001·Journal of Immunological Methods·A TodorovskaP J Hudson
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Feb 24, 2016·Protein Engineering, Design & Selection : PEDS·Teruyo Ojima-KatoHideo Nakano
Nov 3, 2010·Biomedicine & Pharmacotherapy = Biomédecine & Pharmacothérapie·Yan GeXue-Guang Zhang

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