Matches in SemOpenAlex for { <https://semopenalex.org/work/W2980474253> ?p ?o ?g. }
Showing items 1 to 77 of
77
with 100 items per page.
- W2980474253 abstract "Phage display bypasses the immune response by bringing the entire immune repertoire to a target antigen, in vitro. The coupling of antibody phage display and next generation sequencing (NGS) has the potential to revolutionise the discovery of novel antibodies. In this study, next generation antibody phage display (NGAPD) strategies were devised to identify novel antibodies against conformational epitopes on the enzyme phytase, and against the hapten phytic acid. These were notably difficult targets and previous immunisation attempts for phytic acid have not demonstrated any immune response for hybridoma or the production of immune phage display libraries. This study used naive human scFv and camelid VHH phage display libraries and it was therefore more difficult to identify potential binders due to a lack of bias towards the targets within a high level of antibody diversity. Following phage panning, the read depth provided by NGS facilitates the identification of novel ligands from within a diverse sub-library, and thus the study also aimed to optimise the use of NGS with naive phage display libraries. The NGS approach was used in comparison with conventional colony picking. The colony picking approach successfully identified phage-fusion binders against the three targets, however when expressed as antibody-MBP fusions the binding activities were lost. The NGS data analysis involved ranking candidates using a Z score followed by further, highly stringent sorting analyses designed to remove nonspecific ligands. Major challenges involved rescuing scFv candidates from their sub-libraries; this was later resolved with the use of the camelid VHH library that uses a single antibody framework. The NGAPD concluded with a range of NGS candidates synthesised from the final VHH experiment. The candidates were cloned in a single step into a pMAL expression system using inverse PCR. Finally, the NGS candidates were expressed as MBP fusions; however no binding activity was detected. The final section of the study considered alternative bioinformatics analysis strategies for NGS data that have been used in the literature for both antibodies and peptides and also suggested novel enrichment profiles as a means of identifying selection dependant ligands. The VHH NGS data was reassessed based solely on candidate frequency and enrichment throughout panning. A selection of antibody candidates were identified by both bioinformatics methods for phytase, suggesting that the original NGS analysis strategy was successfully identifying promising candidates that were more likely to be conformation specific." @default.
- W2980474253 created "2019-10-25" @default.
- W2980474253 creator A5037545905 @default.
- W2980474253 date "2019-07-31" @default.
- W2980474253 modified "2023-09-27" @default.
- W2980474253 title "Isolating novel antibodies against difficult targets using next generation antibody phage display" @default.
- W2980474253 hasPublicationYear "2019" @default.
- W2980474253 type Work @default.
- W2980474253 sameAs 2980474253 @default.
- W2980474253 citedByCount "0" @default.
- W2980474253 crossrefType "dissertation" @default.
- W2980474253 hasAuthorship W2980474253A5037545905 @default.
- W2980474253 hasConcept C10079403 @default.
- W2980474253 hasConcept C104317684 @default.
- W2980474253 hasConcept C108944566 @default.
- W2980474253 hasConcept C124913957 @default.
- W2980474253 hasConcept C151730666 @default.
- W2980474253 hasConcept C15336307 @default.
- W2980474253 hasConcept C159654299 @default.
- W2980474253 hasConcept C167625842 @default.
- W2980474253 hasConcept C186268636 @default.
- W2980474253 hasConcept C195616568 @default.
- W2980474253 hasConcept C2776441376 @default.
- W2980474253 hasConcept C2776471321 @default.
- W2980474253 hasConcept C2910693471 @default.
- W2980474253 hasConcept C29144244 @default.
- W2980474253 hasConcept C54355233 @default.
- W2980474253 hasConcept C547475151 @default.
- W2980474253 hasConcept C70721500 @default.
- W2980474253 hasConcept C86803240 @default.
- W2980474253 hasConcept C9086966 @default.
- W2980474253 hasConceptScore W2980474253C10079403 @default.
- W2980474253 hasConceptScore W2980474253C104317684 @default.
- W2980474253 hasConceptScore W2980474253C108944566 @default.
- W2980474253 hasConceptScore W2980474253C124913957 @default.
- W2980474253 hasConceptScore W2980474253C151730666 @default.
- W2980474253 hasConceptScore W2980474253C15336307 @default.
- W2980474253 hasConceptScore W2980474253C159654299 @default.
- W2980474253 hasConceptScore W2980474253C167625842 @default.
- W2980474253 hasConceptScore W2980474253C186268636 @default.
- W2980474253 hasConceptScore W2980474253C195616568 @default.
- W2980474253 hasConceptScore W2980474253C2776441376 @default.
- W2980474253 hasConceptScore W2980474253C2776471321 @default.
- W2980474253 hasConceptScore W2980474253C2910693471 @default.
- W2980474253 hasConceptScore W2980474253C29144244 @default.
- W2980474253 hasConceptScore W2980474253C54355233 @default.
- W2980474253 hasConceptScore W2980474253C547475151 @default.
- W2980474253 hasConceptScore W2980474253C70721500 @default.
- W2980474253 hasConceptScore W2980474253C86803240 @default.
- W2980474253 hasConceptScore W2980474253C9086966 @default.
- W2980474253 hasLocation W29804742531 @default.
- W2980474253 hasOpenAccess W2980474253 @default.
- W2980474253 hasPrimaryLocation W29804742531 @default.
- W2980474253 hasRelatedWork W1964396306 @default.
- W2980474253 hasRelatedWork W1981013063 @default.
- W2980474253 hasRelatedWork W2015863036 @default.
- W2980474253 hasRelatedWork W2045482566 @default.
- W2980474253 hasRelatedWork W2072576950 @default.
- W2980474253 hasRelatedWork W2120823001 @default.
- W2980474253 hasRelatedWork W2124559972 @default.
- W2980474253 hasRelatedWork W2410221116 @default.
- W2980474253 hasRelatedWork W2604025583 @default.
- W2980474253 hasRelatedWork W2608487216 @default.
- W2980474253 hasRelatedWork W2727865656 @default.
- W2980474253 hasRelatedWork W2749679400 @default.
- W2980474253 hasRelatedWork W2767788992 @default.
- W2980474253 hasRelatedWork W2790876771 @default.
- W2980474253 hasRelatedWork W2792807736 @default.
- W2980474253 hasRelatedWork W2799534135 @default.
- W2980474253 hasRelatedWork W2891666054 @default.
- W2980474253 hasRelatedWork W2903771374 @default.
- W2980474253 hasRelatedWork W2970957853 @default.
- W2980474253 hasRelatedWork W3116262273 @default.
- W2980474253 isParatext "false" @default.
- W2980474253 isRetracted "false" @default.
- W2980474253 magId "2980474253" @default.
- W2980474253 workType "dissertation" @default.