Plasmodium falciparum serology: A comparison of two protein production methods for analysis of antibody responses by protein microarray.

The evaluation of protein antigens as putative serologic biomarkers of infection has increasingly shifted to high-throughput, multiplex approaches such as the protein microarray. In vitro transcription/translation (IVTT) systems-a similarly high-throughput protein expression method-are already widel...

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Main Authors: Tate Oulton, Joshua Obiero, Isabel Rodriguez, Isaac Ssewanyana, Rebecca A Dabbs, Christine M Bachman, Bryan Greenhouse, Chris Drakeley, Phil L Felgner, Will Stone, Kevin K A Tetteh
Format: Article
Language:English
Published: Public Library of Science (PLoS) 2022-01-01
Series:PLoS ONE
Online Access:https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0273106&type=printable
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author Tate Oulton
Joshua Obiero
Isabel Rodriguez
Isaac Ssewanyana
Rebecca A Dabbs
Christine M Bachman
Bryan Greenhouse
Chris Drakeley
Phil L Felgner
Will Stone
Kevin K A Tetteh
author_facet Tate Oulton
Joshua Obiero
Isabel Rodriguez
Isaac Ssewanyana
Rebecca A Dabbs
Christine M Bachman
Bryan Greenhouse
Chris Drakeley
Phil L Felgner
Will Stone
Kevin K A Tetteh
author_sort Tate Oulton
collection DOAJ
description The evaluation of protein antigens as putative serologic biomarkers of infection has increasingly shifted to high-throughput, multiplex approaches such as the protein microarray. In vitro transcription/translation (IVTT) systems-a similarly high-throughput protein expression method-are already widely utilised in the production of protein microarrays, though purified recombinant proteins derived from more traditional whole cell based expression systems also play an important role in biomarker characterisation. Here we have performed a side-by-side comparison of antigen-matched protein targets from an IVTT and purified recombinant system, on the same protein microarray. The magnitude and range of antibody responses to purified recombinants was found to be greater than that of IVTT proteins, and responses between targets from different expression systems did not clearly correlate. However, responses between amino acid sequence-matched targets from each expression system were more closely correlated. Despite the lack of a clear correlation between antigen-matched targets produced in each expression system, our data indicate that protein microarrays produced using either method can be used confidently, in a context dependent manner, though care should be taken when comparing data derived from contrasting approaches.
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publisher Public Library of Science (PLoS)
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spelling doaj-art-5061e8e9fd41449b8f4ce5889d16bc642025-08-20T02:31:38ZengPublic Library of Science (PLoS)PLoS ONE1932-62032022-01-01178e027310610.1371/journal.pone.0273106Plasmodium falciparum serology: A comparison of two protein production methods for analysis of antibody responses by protein microarray.Tate OultonJoshua ObieroIsabel RodriguezIsaac SsewanyanaRebecca A DabbsChristine M BachmanBryan GreenhouseChris DrakeleyPhil L FelgnerWill StoneKevin K A TettehThe evaluation of protein antigens as putative serologic biomarkers of infection has increasingly shifted to high-throughput, multiplex approaches such as the protein microarray. In vitro transcription/translation (IVTT) systems-a similarly high-throughput protein expression method-are already widely utilised in the production of protein microarrays, though purified recombinant proteins derived from more traditional whole cell based expression systems also play an important role in biomarker characterisation. Here we have performed a side-by-side comparison of antigen-matched protein targets from an IVTT and purified recombinant system, on the same protein microarray. The magnitude and range of antibody responses to purified recombinants was found to be greater than that of IVTT proteins, and responses between targets from different expression systems did not clearly correlate. However, responses between amino acid sequence-matched targets from each expression system were more closely correlated. Despite the lack of a clear correlation between antigen-matched targets produced in each expression system, our data indicate that protein microarrays produced using either method can be used confidently, in a context dependent manner, though care should be taken when comparing data derived from contrasting approaches.https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0273106&type=printable
spellingShingle Tate Oulton
Joshua Obiero
Isabel Rodriguez
Isaac Ssewanyana
Rebecca A Dabbs
Christine M Bachman
Bryan Greenhouse
Chris Drakeley
Phil L Felgner
Will Stone
Kevin K A Tetteh
Plasmodium falciparum serology: A comparison of two protein production methods for analysis of antibody responses by protein microarray.
PLoS ONE
title Plasmodium falciparum serology: A comparison of two protein production methods for analysis of antibody responses by protein microarray.
title_full Plasmodium falciparum serology: A comparison of two protein production methods for analysis of antibody responses by protein microarray.
title_fullStr Plasmodium falciparum serology: A comparison of two protein production methods for analysis of antibody responses by protein microarray.
title_full_unstemmed Plasmodium falciparum serology: A comparison of two protein production methods for analysis of antibody responses by protein microarray.
title_short Plasmodium falciparum serology: A comparison of two protein production methods for analysis of antibody responses by protein microarray.
title_sort plasmodium falciparum serology a comparison of two protein production methods for analysis of antibody responses by protein microarray
url https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0273106&type=printable
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