A novel therapeutic antibody screening method using bacterial high-content imaging reveals functional antibody binding phenotypes of Escherichia coli ST131

Autor: Stephen Baker, Catherine Ludden, Zoe A. Dyson, Stephen Reece, David Goulding, Josefin Bartholdson Scott, Gordon Dougan, Sarah E. Smith, Mailis Maes, Paul Kellam
Přispěvatelé: Baker, Stephen [0000-0003-1308-5755], Dougan, Gordon [0000-0003-0022-965X], Bartholdson Scott, Josefin [0000-0003-3380-4446], Apollo - University of Cambridge Repository
Rok vydání: 2020
Předmět:
0301 basic medicine
lcsh:Medicine
medicine.disease_cause
Antimicrobial resistance
631/154/51/1568
631/154/1435/2417
Insertion sequence
lcsh:Science
631/326
Escherichia coli Infections
Phylogeny
Multidisciplinary
biology
Virulence
Chemistry
631/326/41
High-throughput screening
article
O Antigens
Phenotype
Anti-Bacterial Agents
Antibody
Pathogens
Phenotypic screening
medicine.drug_class
030106 microbiology
Microbial Sensitivity Tests
Monoclonal antibody
Antibodies
Monoclonal
Humanized
Microbiology
Polymorphism
Single Nucleotide
03 medical and health sciences
Drug Resistance
Bacterial
medicine
Escherichia coli
Humans
Gene
Bacteria
631/326/421
lcsh:R
High-Throughput Screening Assays
Interspersed Repetitive Sequences
Microscopy
Electron
030104 developmental biology
631/326/22/1434
biology.protein
Feasibility Studies
lcsh:Q
Antibody therapy
631/154/1435/2163
Zdroj: Scientific Reports, Vol 10, Iss 1, Pp 1-13 (2020)
Scientific Reports
Popis: Funder: National Institute for Health Research; doi: http://dx.doi.org/10.13039/501100000272
Funder: MRC Proximity to Discovery: Industry Engagement Fund Biomedical Research Exchange Programme
The increase of antimicrobial resistance (AMR), and lack of new classes of licensed antimicrobials, have made alternative treatment options for AMR pathogens increasingly attractive. Recent studies have demonstrated anti-bacterial efficacy of a humanised monoclonal antibody (mAb) targeting the O25b O-antigen of Escherichia coli ST131. To evaluate the phenotypic effects of antibody binding to diverse clinical E. coli ST131 O25b bacterial isolates in high-throughput, we designed a novel mAb screening method using high-content imaging (HCI) and image-based morphological profiling to screen a mAb targeting the O25b O-antigen. Screening the antibody against a panel of 86 clinical E. coli ST131 O25:H4 isolates revealed 4 binding phenotypes: no binding (18.60%), weak binding (4.65%), strong binding (69.77%) and strong agglutinating binding (6.98%). Impaired antibody binding could be explained by the presence of insertion sequences or mutations in O-antigen or lipopolysaccharide core biosynthesis genes, affecting the amount, structure or chain length of the O-antigen. The agglutinating binding phenotype was linked with lower O-antigen density, enhanced antibody-mediated phagocytosis and increased serum susceptibly. This study highlights the need to screen candidate mAbs against large panels of clinically relevant isolates, and that HCI can be used to evaluate mAb binding affinity and potential functional efficacy against AMR bacteria.
Databáze: OpenAIRE
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