Development and validation of a high throughput system for discovery of antigens for autoantibody detection.

An assay employing a panel of tumor-associated antigens has been validated and is available commercially (EarlyCDT®-Lung) to aid the early detection of lung cancer by measurement of serum autoantibodies. The high throughput (HTP) strategy described herein was pursued to identify new antigens to add...

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Autores principales:	Isabel K Macdonald, Jared Allen, Andrea Murray, Celine B Parsy-Kowalska, Graham F Healey, Caroline J Chapman, Herbert F Sewell, John F R Robertson
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2012
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/30b6bdc9b68c4fcf9e3b714380df206a
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spelling	oai:doaj.org-article:30b6bdc9b68c4fcf9e3b714380df206a2021-11-18T07:12:02ZDevelopment and validation of a high throughput system for discovery of antigens for autoantibody detection.1932-620310.1371/journal.pone.0040759https://doaj.org/article/30b6bdc9b68c4fcf9e3b714380df206a2012-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/22815807/pdf/?tool=EBIhttps://doaj.org/toc/1932-6203An assay employing a panel of tumor-associated antigens has been validated and is available commercially (EarlyCDT®-Lung) to aid the early detection of lung cancer by measurement of serum autoantibodies. The high throughput (HTP) strategy described herein was pursued to identify new antigens to add to the EarlyCDT-Lung panel and to assist in the development of new panels for other cancers. Two ligation-independent cloning vectors were designed and synthesized, producing fusion proteins suitable for the autoantibody ELISA. We developed an abridged HTP version of the validated autoantibody ELISA, determining that results reflected the performance of the EarlyCDT assay, by comparing results on both formats. Once validated this HTP ELISA was utilized to screen multiple fusion proteins prepared on small-scale, by a HTP expression screen. We determined whether the assay performance for these HTP protein batches was an accurate reflection of the performance of R&D or commercial batches. A HTP discovery platform for the identification and optimal production of tumor-associated antigens which detects autoantibodies has been developed and validated. The most favorable conditions for the exposure of immunogenic epitopes were assessed to produce discriminatory proteins for use in a commercial ELISA. This process is rapid and cost-effective compared to standard cloning and screening technologies and enables rapid advancement in the field of autoantibody assay discovery. This approach will significantly reduce timescale and costs for developing similar panels of autoantibody assays for the detection of other cancer types with the ultimate aim of improved overall survival due to early diagnosis and treatment.Isabel K MacdonaldJared AllenAndrea MurrayCeline B Parsy-KowalskaGraham F HealeyCaroline J ChapmanHerbert F SewellJohn F R RobertsonPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 7, Iss 7, p e40759 (2012)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Isabel K Macdonald Jared Allen Andrea Murray Celine B Parsy-Kowalska Graham F Healey Caroline J Chapman Herbert F Sewell John F R Robertson Development and validation of a high throughput system for discovery of antigens for autoantibody detection.
description	An assay employing a panel of tumor-associated antigens has been validated and is available commercially (EarlyCDT®-Lung) to aid the early detection of lung cancer by measurement of serum autoantibodies. The high throughput (HTP) strategy described herein was pursued to identify new antigens to add to the EarlyCDT-Lung panel and to assist in the development of new panels for other cancers. Two ligation-independent cloning vectors were designed and synthesized, producing fusion proteins suitable for the autoantibody ELISA. We developed an abridged HTP version of the validated autoantibody ELISA, determining that results reflected the performance of the EarlyCDT assay, by comparing results on both formats. Once validated this HTP ELISA was utilized to screen multiple fusion proteins prepared on small-scale, by a HTP expression screen. We determined whether the assay performance for these HTP protein batches was an accurate reflection of the performance of R&D or commercial batches. A HTP discovery platform for the identification and optimal production of tumor-associated antigens which detects autoantibodies has been developed and validated. The most favorable conditions for the exposure of immunogenic epitopes were assessed to produce discriminatory proteins for use in a commercial ELISA. This process is rapid and cost-effective compared to standard cloning and screening technologies and enables rapid advancement in the field of autoantibody assay discovery. This approach will significantly reduce timescale and costs for developing similar panels of autoantibody assays for the detection of other cancer types with the ultimate aim of improved overall survival due to early diagnosis and treatment.
format	article
author	Isabel K Macdonald Jared Allen Andrea Murray Celine B Parsy-Kowalska Graham F Healey Caroline J Chapman Herbert F Sewell John F R Robertson
author_facet	Isabel K Macdonald Jared Allen Andrea Murray Celine B Parsy-Kowalska Graham F Healey Caroline J Chapman Herbert F Sewell John F R Robertson
author_sort	Isabel K Macdonald
title	Development and validation of a high throughput system for discovery of antigens for autoantibody detection.
title_short	Development and validation of a high throughput system for discovery of antigens for autoantibody detection.
title_full	Development and validation of a high throughput system for discovery of antigens for autoantibody detection.
title_fullStr	Development and validation of a high throughput system for discovery of antigens for autoantibody detection.
title_full_unstemmed	Development and validation of a high throughput system for discovery of antigens for autoantibody detection.
title_sort	development and validation of a high throughput system for discovery of antigens for autoantibody detection.
publisher	Public Library of Science (PLoS)
publishDate	2012
url	https://doaj.org/article/30b6bdc9b68c4fcf9e3b714380df206a
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Development and validation of a high throughput system for discovery of antigens for autoantibody detection.

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