9 research outputs found

    Monoclonal Human Antibodies That Recognise the Exposed N and C Terminal Regions of the Often-Overlooked SARS-CoV-2 ORF3a Transmembrane Protein

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    Acknowledgments: The authors would like to gratefully acknowledge the efforts of Julia Martinez Fraile, Richard Lofthouse, Lewis Penny, Mohammad Arastoo and Natalia Cattelan for providing training and assistance with various experimental procedures described in this study. The authors also thank the University of Aberdeen Microscopy and Histology Facility for training and access to fluorescence microscopy and Aberdeen Proteomics for access to BiacoreX100 for SPR binding analysis. Funding: Chief Scientist Office, Scottish Government (COV/ABN/20/01). MRC Centre for Medical Mycology at the University of Exeter (MR/P501955/2).Peer reviewedPublisher PD

    Current Progress and Future Directions for Tau-Based Fluid Biomarker Diagnostics in Alzheimer’s Disease

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    Funding: Research activity relating to tau-based immunodiagnostics performed by the Scottish Biologics Facility was funded by Genting TauRx Diagnostic Centre Sdn. Bhd. Acknowledgments: Figures were created using www.BioRender.com.Peer reviewedPublisher PD

    Monoclonal antibodies targeting surface exposed epitopes of Candida albicans cell wall proteins confer in vivo protection in an infection model

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    ACKNOWLEDGMENTS We gratefully acknowledge Kevin McKenzie and Lucy Wight from the University of Aberdeen Microscopy and Histology Facility for training and access to fluorescence microscopy and for their support and assistance in this work. We also gratefully acknowledge David Stead from Aberdeen Proteomics for his support and assistance with the Candida proteome analysis and the staff of the University of Aberdeen Medical Research Facility for their support and assistance with the mouse studies. This work was supported by the following research grants: the High Throughput and Fragment Screening Fund, Scottish Universities Life Sciences Alliance (SULSA); a seed corn award from the University of Aberdeen Wellcome Trust Institutional Strategic Support Fund; an M.Res. studentship by the Medical Research Council Centre for Medical Mycology at the University of Aberdeen (grant number MR/P501955/1); a Ph.D. studentship from the Institute of Medical Sciences, University of Aberdeen; a Ph.D. studentship from Taibah University and a Saudi Government scholarship; and a Ph.D. studentship by the European Union’s Horizon 2020 research and innovation program under Marie Sklodowska-Curie grant agreement number H2020-MSCA-ITN-2014-642095 (OPATHY). C.A.M., S.P., and A.J.P. contributed to the concept and study design. C.A.M. and S.P. developed the methodology. S.A.A. and L.F. performed recombinant antibody generation and ELISAs. L.F. and M.M. completed IgG reformatting and the production of mAbs for animal studies. M.M., T.H.T., and L.A.W. performed ELISAs and immunofluorescence staining. M.M. performed macrophage assays, and D.M.M. planned, conducted, and analyzed animal studies. C.A.M., S.P., and A.J.P. contributed to funding acquisition and project administration, and C.A.M., S.P., and L.A.W. contributed to the supervision and training of M.Res. and Ph.D. students. S.P. wrote the original draft, and C.A.M., D.M.M., and A.J.P. completed review and editing. All authors had full access to the data and approved the manuscript before it was submitted by the corresponding author(s). S.P., A.J.P., and C.A.M. are inventors on a patent related to the development of antifungal antibodies to surface-exposed epitopes of fungal pathogens owned by the University of Aberdeen. All other authors declare that they have no competing interests.Peer reviewedPublisher PD

    Disruption of Pseudomonas aeruginosa quorum sensing using high-sensitivity phage antibodies derived from immunised sheep

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    Pseudomonas aeruginosa is an opportunistic human pathogen which, like many other Gram negative pathogens, employs quorum sensing - regulated virulence factors to establish an infection in its host. Quorum sensing in P. aeruginosa populations is controlled by low molecular weight (hapten) signalling molecules known as homoserine lactones (HSLs). Blocking bacterial communication using antibodies is an attractive strategy for infection control as QS takes a central role in P. aeruginosa infections, and antibodies can recognise their targets with exquisite specificity. There are two well-studied QS circuits in P. aeruginosa- the Las system and the Rhl system, controlled by two autoinducers compounds, 3-oxo-C12-HSL and C4-HSL respectively. Antibodies raised against HSL compounds can reduce the expression of virulence factors controlled by QS circuit and the immunomodulatory effects of 3- oxo-C12-HSL. In order to generate antibodies with high sensitivities against the autoinducer compounds of P. aeruginosa, a panel of HSL compounds was synthesised, conjugated to the carrier protein and used for sheep immunisation. High specificity anti-HSL antibodies were isolated from an immunised sheep antibody repertoire using phage display technology. These phage antibody hits were converted into single chain antibody (scAb) format, which possessed a HuCκ gene for detection and 6x histidine tag for purification. Soluble scAbs expressed in E. coli were purified and characterised using ELISA. Unique clones showing high sensitivity for free HSL compounds were reformatted into sheep-mouse chimeric IgGs, expressed transiently in COS 7 cells and characterised using biochemical assays. These cross-reactive monoclonal antibodies were shown to recognise HSL compounds in low nanomolar concentrations and have the potential to reduce virulence gene expression in P. aeruginosa.EThOS - Electronic Theses Online ServiceGBUnited Kingdo
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