Article ID | Journal | Published Year | Pages | File Type |
---|---|---|---|---|
5513653 | Methods | 2017 | 37 Pages |
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.
Keywords
deoxyribonucleaseVλCκjoining segmentJ segmentSOECDR1MOPSIFAScAbTMBCFABCAPBSTIMACHRPPBSIMSscFvIPTG3-(N-morpholino)-propanesulfonic acid3,3′,5,5′-tetramethylbenzidinecomplete Freund’s adjuvantincomplete Freund’s adjuvantDNAseF(ab′)2VκRecombinant antibodysplicing by overlap extensionisopropyl β-D-1-thiogalactopyranosidebicinchoninic acidReverse transcriptaseELISAheavy chainSuper brothSOCFabPhosphate buffered salinesingle chain antibody fragmentsingle-chain variable fragmentlysogeny brothhemagglutininunitpolymerase chain reactionPCRPanningPurificationHorseradish peroxidaseImmobilised metal affinity chromatography
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Authors
H. Ma, R. O'Kennedy,