Imperial College London

ProfessorPaulFreemont

Faculty of MedicineDepartment of Medicine

Chair in Protein Crystallography
 
 
 
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Contact

 

+44 (0)20 7594 5327p.freemont

 
 
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Location

 

259Sir Alexander Fleming BuildingSouth Kensington Campus

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Summary

 

Publications

Citation

BibTex format

@article{Kylilis:2019:10.1021/acssensors.8b01163,
author = {Kylilis, N and Riangrungroj, P and Lai, H-E and Salema, V and Fernández, LÁ and Stan, G-BV and Freemont, PS and Polizzi, KM},
doi = {10.1021/acssensors.8b01163},
journal = {ACS Sens},
pages = {370--378},
title = {Whole-Cell Biosensor with Tunable Limit of Detection Enables Low-Cost Agglutination Assays for Medical Diagnostic Applications.},
url = {http://dx.doi.org/10.1021/acssensors.8b01163},
volume = {4},
year = {2019}
}

RIS format (EndNote, RefMan)

TY  - JOUR
AB - Whole-cell biosensors can form the basis of affordable, easy-to-use diagnostic tests that can be readily deployed for point-of-care (POC) testing, but to date the detection of analytes such as proteins that cannot easily diffuse across the cell membrane has been challenging. Here we developed a novel biosensing platform based on cell agglutination using an E. coli whole-cell biosensor surface-displaying nanobodies which bind selectively to a target protein analyte. As a proof-of-concept, we show the feasibility of this design to detect a model analyte at nanomolar concentrations. Moreover, we show that the design architecture is flexible by building assays optimized to detect a range of model analyte concentrations using straightforward design rules and a mathematical model. Finally, we re-engineer our whole-cell biosensor for the detection of a medically relevant biomarker by the display of two different nanobodies against human fibrinogen and demonstrate a detection limit as low as 10 pM in diluted human plasma. Overall, we demonstrate that our agglutination technology fulfills the requirement of POC testing by combining low-cost nanobody production, customizable detection range and low detection limits. This technology has the potential to produce affordable diagnostics for field-testing in the developing world, emergency or disaster relief sites, as well as routine medical testing and personalized medicine.
AU - Kylilis,N
AU - Riangrungroj,P
AU - Lai,H-E
AU - Salema,V
AU - Fernández,LÁ
AU - Stan,G-BV
AU - Freemont,PS
AU - Polizzi,KM
DO - 10.1021/acssensors.8b01163
EP - 378
PY - 2019///
SP - 370
TI - Whole-Cell Biosensor with Tunable Limit of Detection Enables Low-Cost Agglutination Assays for Medical Diagnostic Applications.
T2 - ACS Sens
UR - http://dx.doi.org/10.1021/acssensors.8b01163
UR - https://www.ncbi.nlm.nih.gov/pubmed/30623662
UR - http://hdl.handle.net/10044/1/65633
VL - 4
ER -