Recombinant antibody fragment production

•Generation of recombinant antibodies using phage display.•Screening of specific recombinant antibodies via panning.•Selection and purification of soluble recombinant antibodies using immobilised metal affinity chromatography (IMAC). Recombinant antibodies are now very important in both therapeutics...

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Bibliographic Details
Published in:Methods (San Diego, Calif.) Calif.), 2017-03, Vol.116, p.23-33
Main Authors: Ma, H., O’Kennedy, R.
Format: Article
Language:English
Subjects:
Animals
Antigens - administration & dosage
Chromatography, Affinity
Cloning, Molecular
Enzyme-Linked Immunosorbent Assay
Escherichia coli - genetics
Escherichia coli - metabolism
Freund's Adjuvant - administration & dosage
Gene Expression
Humans
Mice
Panning
Peptide Library
Purification
Recombinant antibody
Recombinant Proteins - biosynthesis
Recombinant Proteins - blood
Recombinant Proteins - isolation & purification
scFv
Single-Chain Antibodies - biosynthesis
Single-Chain Antibodies - blood
Single-Chain Antibodies - isolation & purification
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Summary:•Generation of recombinant antibodies using phage display.•Screening of specific recombinant antibodies via panning.•Selection and purification of soluble recombinant antibodies using immobilised metal affinity chromatography (IMAC). Recombinant antibodies are now very important in both therapeutics and diagnostics and offer significant advantages over conventional antibodies. The generation of a single-chain variable antibody fragment (scFv) (a common and important recombinant antibody format) is used to demonstrate the construction of a recombinant antibody library. An immunotube-based two-day panning approach, using Escherichia coli as an expression system, is utilised for antibody screening. The methods used for antibody selection and purification using immobilised metal affinity chromatography (IMAC) are described.
ISSN:1046-2023
1095-9130
DOI:10.1016/j.ymeth.2016.11.008