RT Journal Article
T1 Unraveling the Properties of Phage Display Fab Libraries and Their Use in the Selection of Gliadin-Specific Probes by Applying High-Throughput Nanopore Sequencing
A1 García Calvo, Eduardo Rafael
A1 García García, Aina
A1 Rodríguez Gómez, Santiago
A1 Martín De Santos, María Del Rosario
A1 García Lacarra, Teresa
AB Directed evolution is a pivotal strategy for new antibody discovery, which allowed the generation of high-affinity Fabs against gliadin from two antibody libraries in our previous studies. One of the libraries was exclusively derived from celiac patients’ mRNA (immune library) while the other was obtained through a protein engineering approach (semi-immune library). Recent advances in high-throughput DNA sequencing techniques are revolutionizing research across genomics, epigenomics, and transcriptomics. In the present work, an Oxford Nanopore in-lab sequencing device was used to comprehensively characterize the composition of the constructed libraries, both at the beginning and throughout the phage-mediated selection processes against gliadin. A customized analysis pipeline was used to select high-quality reads, annotate chain distribution, perform sequence analysis, and conduct statistical comparisons between the different selection rounds. Some immunological attributes of the most representative phage variants after the selection process were also determined. Sequencing results revealed the successful transfer of the celiac immune response features to the immune library and the antibodies derived from it, suggesting the crucial role of these features in guiding the selection of high-affinity recombinant Fabs against gliadin. In summary, high-throughput DNA sequencing has improved our understanding of the selection processes aimed at generating molecular binders against gliadin.
PB MDPI
SN 1999-4915
YR 2024
FD 2024-04-26
LK https://hdl.handle.net/20.500.14352/104803
UL https://hdl.handle.net/20.500.14352/104803
LA eng
NO Garcia-Calvo, E.; García-García, A.; Rodríguez, S.; Martín, R.; García, T. Unraveling the Properties of Phage Display Fab Libraries and Their Use in the Selection of Gliadin-Specific Probes by Applying High-Throughput Nanopore Sequencing. Viruses 2024, 16, 686. https://doi.org/10.3390/ v16050686
NO Ministerio de Ciencia e Innovación (España)
DS Docta Complutense
RD 6 oct 2024