Determination of variable region sequences from hybridoma immunoglobulins that target Mycobacterium tuberculosis virulence factors.

Mycobacterium tuberculosis (Mtb) infects one-quarter of the world's population. Mtb and HIV coinfections enhance the comorbidity of tuberculosis (TB) and AIDS, accounting for one-third of all AIDS-associated mortalities. Humoral antibody to Mtb correlates with TB susceptibility, and engineering...

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Autores principales:	Hui-Chen Chang Foreman, Andrew Frank, Timothy T Stedman
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/174ae2ca514c4cfeaa8460558c342bb8
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spelling	oai:doaj.org-article:174ae2ca514c4cfeaa8460558c342bb82021-12-02T20:17:40ZDetermination of variable region sequences from hybridoma immunoglobulins that target Mycobacterium tuberculosis virulence factors.1932-620310.1371/journal.pone.0256079https://doaj.org/article/174ae2ca514c4cfeaa8460558c342bb82021-01-01T00:00:00Zhttps://doi.org/10.1371/journal.pone.0256079https://doaj.org/toc/1932-6203Mycobacterium tuberculosis (Mtb) infects one-quarter of the world's population. Mtb and HIV coinfections enhance the comorbidity of tuberculosis (TB) and AIDS, accounting for one-third of all AIDS-associated mortalities. Humoral antibody to Mtb correlates with TB susceptibility, and engineering of Mtb antibodies may lead to new diagnostics and therapeutics. The characterization and validation of functional immunoglobulin (Ig) variable chain (IgV) sequences provide a necessary first step towards developing therapeutic antibodies against pathogens. The virulence-associated Mtb antigens SodA (Superoxide Dismutase), KatG (Catalase), PhoS1/PstS1 (regulatory factor), and GroES (heat shock protein) are potential therapeutic targets but lacked IgV sequence characterization. Putative IgV sequences were identified from the mRNA of hybridomas targeting these antigens and isotype-switched into a common immunoglobulin fragment crystallizable region (Fc region) backbone, subclass IgG2aκ. Antibodies were validated by demonstrating recombinant Ig assembly and secretion, followed by the determination of antigen-binding specificity using ELISA and immunoblot assay.Hui-Chen Chang ForemanAndrew FrankTimothy T StedmanPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 16, Iss 8, p e0256079 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Hui-Chen Chang Foreman Andrew Frank Timothy T Stedman Determination of variable region sequences from hybridoma immunoglobulins that target Mycobacterium tuberculosis virulence factors.
description	Mycobacterium tuberculosis (Mtb) infects one-quarter of the world's population. Mtb and HIV coinfections enhance the comorbidity of tuberculosis (TB) and AIDS, accounting for one-third of all AIDS-associated mortalities. Humoral antibody to Mtb correlates with TB susceptibility, and engineering of Mtb antibodies may lead to new diagnostics and therapeutics. The characterization and validation of functional immunoglobulin (Ig) variable chain (IgV) sequences provide a necessary first step towards developing therapeutic antibodies against pathogens. The virulence-associated Mtb antigens SodA (Superoxide Dismutase), KatG (Catalase), PhoS1/PstS1 (regulatory factor), and GroES (heat shock protein) are potential therapeutic targets but lacked IgV sequence characterization. Putative IgV sequences were identified from the mRNA of hybridomas targeting these antigens and isotype-switched into a common immunoglobulin fragment crystallizable region (Fc region) backbone, subclass IgG2aκ. Antibodies were validated by demonstrating recombinant Ig assembly and secretion, followed by the determination of antigen-binding specificity using ELISA and immunoblot assay.
format	article
author	Hui-Chen Chang Foreman Andrew Frank Timothy T Stedman
author_facet	Hui-Chen Chang Foreman Andrew Frank Timothy T Stedman
author_sort	Hui-Chen Chang Foreman
title	Determination of variable region sequences from hybridoma immunoglobulins that target Mycobacterium tuberculosis virulence factors.
title_short	Determination of variable region sequences from hybridoma immunoglobulins that target Mycobacterium tuberculosis virulence factors.
title_full	Determination of variable region sequences from hybridoma immunoglobulins that target Mycobacterium tuberculosis virulence factors.
title_fullStr	Determination of variable region sequences from hybridoma immunoglobulins that target Mycobacterium tuberculosis virulence factors.
title_full_unstemmed	Determination of variable region sequences from hybridoma immunoglobulins that target Mycobacterium tuberculosis virulence factors.
title_sort	determination of variable region sequences from hybridoma immunoglobulins that target mycobacterium tuberculosis virulence factors.
publisher	Public Library of Science (PLoS)
publishDate	2021
url	https://doaj.org/article/174ae2ca514c4cfeaa8460558c342bb8
work_keys_str_mv	AT huichenchangforeman determinationofvariableregionsequencesfromhybridomaimmunoglobulinsthattargetmycobacteriumtuberculosisvirulencefactors AT andrewfrank determinationofvariableregionsequencesfromhybridomaimmunoglobulinsthattargetmycobacteriumtuberculosisvirulencefactors AT timothytstedman determinationofvariableregionsequencesfromhybridomaimmunoglobulinsthattargetmycobacteriumtuberculosisvirulencefactors
_version_	1718374329767428096

Determination of variable region sequences from hybridoma immunoglobulins that target Mycobacterium tuberculosis virulence factors.

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