Cargando…

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...

Descripción completa

Detalles Bibliográficos
Autores principales: Oulton, Tate, Obiero, Joshua, Rodriguez, Isabel, Ssewanyana, Isaac, Dabbs, Rebecca A., Bachman, Christine M., Greenhouse, Bryan, Drakeley, Chris, Felgner, Phil L., Stone, Will, Tetteh, Kevin K. A.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9423672/
https://www.ncbi.nlm.nih.gov/pubmed/36037183
http://dx.doi.org/10.1371/journal.pone.0273106
_version_ 1784778071915626496
author Oulton, Tate
Obiero, Joshua
Rodriguez, Isabel
Ssewanyana, Isaac
Dabbs, Rebecca A.
Bachman, Christine M.
Greenhouse, Bryan
Drakeley, Chris
Felgner, Phil L.
Stone, Will
Tetteh, Kevin K. A.
author_facet Oulton, Tate
Obiero, Joshua
Rodriguez, Isabel
Ssewanyana, Isaac
Dabbs, Rebecca A.
Bachman, Christine M.
Greenhouse, Bryan
Drakeley, Chris
Felgner, Phil L.
Stone, Will
Tetteh, Kevin K. A.
author_sort Oulton, Tate
collection PubMed
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.
format Online
Article
Text
id pubmed-9423672
institution National Center for Biotechnology Information
language English
publishDate 2022
publisher Public Library of Science
record_format MEDLINE/PubMed
spelling pubmed-94236722022-08-30 Plasmodium falciparum serology: A comparison of two protein production methods for analysis of antibody responses by protein microarray Oulton, Tate Obiero, Joshua Rodriguez, Isabel Ssewanyana, Isaac Dabbs, Rebecca A. Bachman, Christine M. Greenhouse, Bryan Drakeley, Chris Felgner, Phil L. Stone, Will Tetteh, Kevin K. A. PLoS One Research Article 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. Public Library of Science 2022-08-29 /pmc/articles/PMC9423672/ /pubmed/36037183 http://dx.doi.org/10.1371/journal.pone.0273106 Text en © 2022 Oulton et al https://creativecommons.org/licenses/by/4.0/This is an open access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
spellingShingle Research Article
Oulton, Tate
Obiero, Joshua
Rodriguez, Isabel
Ssewanyana, Isaac
Dabbs, Rebecca A.
Bachman, Christine M.
Greenhouse, Bryan
Drakeley, Chris
Felgner, Phil L.
Stone, Will
Tetteh, Kevin K. A.
Plasmodium falciparum serology: A comparison of two protein production methods for analysis of antibody responses by protein microarray
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
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9423672/
https://www.ncbi.nlm.nih.gov/pubmed/36037183
http://dx.doi.org/10.1371/journal.pone.0273106
work_keys_str_mv AT oultontate plasmodiumfalciparumserologyacomparisonoftwoproteinproductionmethodsforanalysisofantibodyresponsesbyproteinmicroarray
AT obierojoshua plasmodiumfalciparumserologyacomparisonoftwoproteinproductionmethodsforanalysisofantibodyresponsesbyproteinmicroarray
AT rodriguezisabel plasmodiumfalciparumserologyacomparisonoftwoproteinproductionmethodsforanalysisofantibodyresponsesbyproteinmicroarray
AT ssewanyanaisaac plasmodiumfalciparumserologyacomparisonoftwoproteinproductionmethodsforanalysisofantibodyresponsesbyproteinmicroarray
AT dabbsrebeccaa plasmodiumfalciparumserologyacomparisonoftwoproteinproductionmethodsforanalysisofantibodyresponsesbyproteinmicroarray
AT bachmanchristinem plasmodiumfalciparumserologyacomparisonoftwoproteinproductionmethodsforanalysisofantibodyresponsesbyproteinmicroarray
AT greenhousebryan plasmodiumfalciparumserologyacomparisonoftwoproteinproductionmethodsforanalysisofantibodyresponsesbyproteinmicroarray
AT drakeleychris plasmodiumfalciparumserologyacomparisonoftwoproteinproductionmethodsforanalysisofantibodyresponsesbyproteinmicroarray
AT felgnerphill plasmodiumfalciparumserologyacomparisonoftwoproteinproductionmethodsforanalysisofantibodyresponsesbyproteinmicroarray
AT stonewill plasmodiumfalciparumserologyacomparisonoftwoproteinproductionmethodsforanalysisofantibodyresponsesbyproteinmicroarray
AT tettehkevinka plasmodiumfalciparumserologyacomparisonoftwoproteinproductionmethodsforanalysisofantibodyresponsesbyproteinmicroarray