Imperial College London
Alternate

ProfessorRobinShattock

Faculty of Medicine, Department of Infectious Disease

Chair in Mucosal Infection and Immunity
 
 
 
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Contact

 

+44 (0)20 7594 5206r.shattock

 
 
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Location

 

453Wright Fleming WingSt Mary's Campus

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Summary

 

Publications

Citation

BibTex format

@article{Pinder:2017:10.4049/jimmunol.1701253,
author = {Pinder, CL and kratochvil, S and Cizmeci, D and Muir, L and Guo, Y and Shattock, R and McKay, PF},
doi = {10.4049/jimmunol.1701253},
journal = {Journal of Immunology},
title = {Isolation and Characterization of Antigen-Specic Plasmablasts Using a Novel Flow Cytometry–Based Ig Capture Assay},
url = {http://dx.doi.org/10.4049/jimmunol.1701253},
year = {2017}
}
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RIS format (EndNote, RefMan)

TY  - JOUR
AB  - We report the development of a novel flow cytometry–based Ig capture assay (ICA) for the identification and sorting of individual Ab-secreting cells based on their Ag reactivity. The ICA represents a fast and versatile tool for single-cell sorting of peripheral plasmablasts, streamlining subsequent Ab analysis, and cloning. We demonstrate the utility of the assay by isolating Ag-reactive plasmablasts from cryopreserved PBMC obtained from volunteers vaccinated with a recombinant HIV envelope protein. To show the specificity of the ICA, we produced Ag-specific Abs from these cells and subsequently verified their Ag reactivity via ELISA. Furthermore, we used the ICA to track Ag-specific plasmablast responses in HIV-vaccine recipients over a period of 42 d and performed a head-to-head comparison with a conventional B cell ELISpot. Results were highly comparable, highlighting that this assay is a viable alternative for monitoring Ag-specific plasmablast responses at early time points after infection or vaccination. The ICA provides important added benefits in that phenotypic information can be obtained from the identified Ag-specific cells that can then be captured for downstream applications such as B cell sequencing and/or Ab cloning. We envisage the ICA as being a useful tool in Ab repertoire analysis for future clinical trials.
AU  - Pinder,CL
AU  - kratochvil,S
AU  - Cizmeci,D
AU  - Muir,L
AU  - Guo,Y
AU  - Shattock,R
AU  - McKay,PF
DO  - 10.4049/jimmunol.1701253
PY  - 2017///
SN  - 1550-6606
TI  - Isolation and Characterization of Antigen-Specic Plasmablasts Using a Novel Flow Cytometry–Based Ig Capture Assay
T2  - Journal of Immunology
UR  - http://dx.doi.org/10.4049/jimmunol.1701253
ER  -
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