Selection and characterisation of Affimers specific for CEA recognition

Abstract Carcinoembryonic antigen (CEA) is the only blood based protein biomarker at present, used for preoperative screening of advanced colorectal cancer (CRC) patients to determine the appropriate curative treatments and post-surveillance screening for tumour recurrence. Current diagnostics for C...

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Autores principales:	Shazana Hilda Shamsuddin, David G. Jayne, Darren C. Tomlinson, Michael J. McPherson, Paul A. Millner
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/1a33adcba2a44a18b93a84d48aebf2a6
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spelling	oai:doaj.org-article:1a33adcba2a44a18b93a84d48aebf2a62021-12-02T14:01:21ZSelection and characterisation of Affimers specific for CEA recognition10.1038/s41598-020-80354-62045-2322https://doaj.org/article/1a33adcba2a44a18b93a84d48aebf2a62021-01-01T00:00:00Zhttps://doi.org/10.1038/s41598-020-80354-6https://doaj.org/toc/2045-2322Abstract Carcinoembryonic antigen (CEA) is the only blood based protein biomarker at present, used for preoperative screening of advanced colorectal cancer (CRC) patients to determine the appropriate curative treatments and post-surveillance screening for tumour recurrence. Current diagnostics for CRC detection have several limitations and development of a highly sensitive, specific and rapid diagnostic device is required. The majority of such devices developed to date are antibody-based and suffer from shortcomings including multimeric binding, cost and difficulties in mass production. To circumvent antibody-derived limitations, the present study focused on the development of Affimer proteins as a novel alternative binding reagent for CEA detection. Here, we describe the selection, from a phage display library, of Affimers specific to CEA protein. Characterization of three anti-CEA Affimers reveal that these bind specifically and selectively to protein epitopes of CEA from cell culture lysate and on fixed cells. Kinetic binding analysis by SPR show that the Affimers bind to CEA with high affinity and within the nM range. Therefore, they have substantial potential for used as novel affinity reagents in diagnostic imaging, targeted CRC therapy, affinity purification and biosensor applications.Shazana Hilda ShamsuddinDavid G. JayneDarren C. TomlinsonMichael J. McPhersonPaul A. MillnerNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-10 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Shazana Hilda Shamsuddin David G. Jayne Darren C. Tomlinson Michael J. McPherson Paul A. Millner Selection and characterisation of Affimers specific for CEA recognition
description	Abstract Carcinoembryonic antigen (CEA) is the only blood based protein biomarker at present, used for preoperative screening of advanced colorectal cancer (CRC) patients to determine the appropriate curative treatments and post-surveillance screening for tumour recurrence. Current diagnostics for CRC detection have several limitations and development of a highly sensitive, specific and rapid diagnostic device is required. The majority of such devices developed to date are antibody-based and suffer from shortcomings including multimeric binding, cost and difficulties in mass production. To circumvent antibody-derived limitations, the present study focused on the development of Affimer proteins as a novel alternative binding reagent for CEA detection. Here, we describe the selection, from a phage display library, of Affimers specific to CEA protein. Characterization of three anti-CEA Affimers reveal that these bind specifically and selectively to protein epitopes of CEA from cell culture lysate and on fixed cells. Kinetic binding analysis by SPR show that the Affimers bind to CEA with high affinity and within the nM range. Therefore, they have substantial potential for used as novel affinity reagents in diagnostic imaging, targeted CRC therapy, affinity purification and biosensor applications.
format	article
author	Shazana Hilda Shamsuddin David G. Jayne Darren C. Tomlinson Michael J. McPherson Paul A. Millner
author_facet	Shazana Hilda Shamsuddin David G. Jayne Darren C. Tomlinson Michael J. McPherson Paul A. Millner
author_sort	Shazana Hilda Shamsuddin
title	Selection and characterisation of Affimers specific for CEA recognition
title_short	Selection and characterisation of Affimers specific for CEA recognition
title_full	Selection and characterisation of Affimers specific for CEA recognition
title_fullStr	Selection and characterisation of Affimers specific for CEA recognition
title_full_unstemmed	Selection and characterisation of Affimers specific for CEA recognition
title_sort	selection and characterisation of affimers specific for cea recognition
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/1a33adcba2a44a18b93a84d48aebf2a6
work_keys_str_mv	AT shazanahildashamsuddin selectionandcharacterisationofaffimersspecificforcearecognition AT davidgjayne selectionandcharacterisationofaffimersspecificforcearecognition AT darrenctomlinson selectionandcharacterisationofaffimersspecificforcearecognition AT michaeljmcpherson selectionandcharacterisationofaffimersspecificforcearecognition AT paulamillner selectionandcharacterisationofaffimersspecificforcearecognition
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Selection and characterisation of Affimers specific for CEA recognition

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