Expression and purification strategies for the production of single-chain antibody and T-cell receptor fragments in E. coli

This work describes protocols for the production of single-chain antibody and T-cell receptor fragments in E. coli. A choice of methods is given for the purification of the recombinant fragments that rely on the use of either immunoaffinity or metal chelate affinity chromatography. The TCR fragments...

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Bibliographic Details
Published in:Molecular biotechnology 1995-12, Vol.4 (3), p.239-245
Main Authors: MOLLOY, P. E, GRAHAM, B. M, CUPIT, P. M, GRANT, S. D, PORTER, A. J. R, CUNNINGHAM, C
Format: Article
Language:English
Subjects:
Biological and medical sciences
Biotechnology
Chromatography, Affinity
Enzyme-Linked Immunosorbent Assay
Escherichia coli - chemistry
Escherichia coli - genetics
Fundamental and applied biological sciences. Psychology
Gene Expression
Genes, Bacterial
Genetic engineering
Genetic technics
Immunoglobulin Fragments - biosynthesis
Immunoglobulin Fragments - chemistry
Immunoglobulin Fragments - isolation & purification
Immunoglobulins - chemistry
Immunoglobulins - isolation & purification
Methods. Procedures. Technologies
Modification of gene expression level
Peptide Fragments - chemistry
Peptide Fragments - isolation & purification
Plasmids - genetics
Protein Structure, Secondary
Receptors, Antigen, T-Cell - biosynthesis
Recombinant Proteins - chemistry
Recombinant Proteins - isolation & purification
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Summary:This work describes protocols for the production of single-chain antibody and T-cell receptor fragments in E. coli. A choice of methods is given for the purification of the recombinant fragments that rely on the use of either immunoaffinity or metal chelate affinity chromatography. The TCR fragments may have to be denatured and refolded before the fragments attain their proper conformation.
ISSN:1073-6085
1559-0305
DOI:10.1007/BF02779017