title: Recombinant antibody fragment production.
abstract: 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.
authors.affiliation: Biomedical Diagnostics Institute, Dublin City University, Dublin 9, Ireland. Electronic address: hui.ma@dcu.ie.
authors.forename: H
authors.initials: H
authors.lastname: Ma
authors.affiliation: Biomedical Diagnostics Institute, Dublin City University, Dublin 9, Ireland.
authors.forename: R
authors.initials: R
authors.lastname: O'Kennedy
mesh_terms: D000818:Animals; D000941:Antigens / Q000008:administration & dosage; D002846:Chromatography, Affinity; D003001:Cloning, Molecular; D004797:Enzyme-Linked Immunosorbent Assay; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D005620:Freund's Adjuvant / Q000008:administration & dosage*; D015870:Gene Expression; D006801:Humans; D051379:Mice; D019151:Peptide Library; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000097:blood / Q000302:isolation & purification; D057127:Single-Chain Antibodies / Q000096:biosynthesis* / Q000097:blood / Q000302:isolation & purification*
chemical_list: D000941:Antigens; D019151:Peptide Library; D011994:Recombinant Proteins; D057127:Single-Chain Antibodies; D005620:Freund's Adjuvant
keywords: Panning; Purification; Recombinant antibody; scFv