A hybridoma-derived monoclonal antibody with high homology to the aberrant myeloma light chain.

The identification of antibody variable regions in the heavy (VH) and light (VL) chains from hybridomas is necessary for the production of recombinant, sequence-defined monoclonal antibodies (mAbs) and antibody derivatives. This process has received renewed attention in light of recent reports of hy...

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Autores principales:	Ghasidit Pornnoppadol, Boya Zhang, Alec A Desai, Anthony Berardi, Henriette A Remmer, Peter M Tessier, Colin F Greineder
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/2422a6f031f8494c9006d47c01d9a2c4
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spelling	oai:doaj.org-article:2422a6f031f8494c9006d47c01d9a2c42021-12-02T20:13:45ZA hybridoma-derived monoclonal antibody with high homology to the aberrant myeloma light chain.1932-620310.1371/journal.pone.0252558https://doaj.org/article/2422a6f031f8494c9006d47c01d9a2c42021-01-01T00:00:00Zhttps://doi.org/10.1371/journal.pone.0252558https://doaj.org/toc/1932-6203The identification of antibody variable regions in the heavy (VH) and light (VL) chains from hybridomas is necessary for the production of recombinant, sequence-defined monoclonal antibodies (mAbs) and antibody derivatives. This process has received renewed attention in light of recent reports of hybridomas having unintended specificities due to the production of non-antigen specific heavy and/or light chains for the intended antigen. Here we report a surprising finding and potential pitfall in variable domain sequencing of an anti-human CD63 hybridoma. We amplified multiple VL genes from the hybridoma cDNA, including the well-known aberrant Sp2/0 myeloma VK and a unique, full-length VL. After finding that the unique VL failed to yield a functional antibody, we discovered an additional full-length sequence with surprising similarity (~95% sequence identify) to the non-translated myeloma kappa chain but with a correction of its key frameshift mutation. Expression of the recombinant mAb confirmed that this highly homologous sequence is the antigen-specific light chain. Our results highlight the complexity of PCR-based cloning of antibody genes and strategies useful for identification of correct sequences.Ghasidit PornnoppadolBoya ZhangAlec A DesaiAnthony BerardiHenriette A RemmerPeter M TessierColin F GreinederPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 16, Iss 10, p e0252558 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Ghasidit Pornnoppadol Boya Zhang Alec A Desai Anthony Berardi Henriette A Remmer Peter M Tessier Colin F Greineder A hybridoma-derived monoclonal antibody with high homology to the aberrant myeloma light chain.
description	The identification of antibody variable regions in the heavy (VH) and light (VL) chains from hybridomas is necessary for the production of recombinant, sequence-defined monoclonal antibodies (mAbs) and antibody derivatives. This process has received renewed attention in light of recent reports of hybridomas having unintended specificities due to the production of non-antigen specific heavy and/or light chains for the intended antigen. Here we report a surprising finding and potential pitfall in variable domain sequencing of an anti-human CD63 hybridoma. We amplified multiple VL genes from the hybridoma cDNA, including the well-known aberrant Sp2/0 myeloma VK and a unique, full-length VL. After finding that the unique VL failed to yield a functional antibody, we discovered an additional full-length sequence with surprising similarity (~95% sequence identify) to the non-translated myeloma kappa chain but with a correction of its key frameshift mutation. Expression of the recombinant mAb confirmed that this highly homologous sequence is the antigen-specific light chain. Our results highlight the complexity of PCR-based cloning of antibody genes and strategies useful for identification of correct sequences.
format	article
author	Ghasidit Pornnoppadol Boya Zhang Alec A Desai Anthony Berardi Henriette A Remmer Peter M Tessier Colin F Greineder
author_facet	Ghasidit Pornnoppadol Boya Zhang Alec A Desai Anthony Berardi Henriette A Remmer Peter M Tessier Colin F Greineder
author_sort	Ghasidit Pornnoppadol
title	A hybridoma-derived monoclonal antibody with high homology to the aberrant myeloma light chain.
title_short	A hybridoma-derived monoclonal antibody with high homology to the aberrant myeloma light chain.
title_full	A hybridoma-derived monoclonal antibody with high homology to the aberrant myeloma light chain.
title_fullStr	A hybridoma-derived monoclonal antibody with high homology to the aberrant myeloma light chain.
title_full_unstemmed	A hybridoma-derived monoclonal antibody with high homology to the aberrant myeloma light chain.
title_sort	hybridoma-derived monoclonal antibody with high homology to the aberrant myeloma light chain.
publisher	Public Library of Science (PLoS)
publishDate	2021
url	https://doaj.org/article/2422a6f031f8494c9006d47c01d9a2c4
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A hybridoma-derived monoclonal antibody with high homology to the aberrant myeloma light chain.

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