Development and characterization of a unique anti‐IgE mouse monoclonal antibody cross‐reactive between human and canine IgE

Abstract Background The efficacy assessment of human anti‐IgE monoclonal antibodies (mAbs) in animal models before clinical trials is hampered due to the lack of cross‐reactivity of anti‐IgE mAbs between species. Objective We developed CRE‐DR (an anti‐dog IgE monoclonal antibody), an anti‐IgE mouse...

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Autores principales:	Akiko Kumagai, Takuya Nara, Mizuho Uematsu, Yoko Kakinuma, Takashi Saito, Kenichi Masuda
Formato:	article
Lenguaje:	EN
Publicado:	Wiley 2021
Materias:	animals epitopes humans IgE \| monoclonal antibody Immunologic diseases. Allergy RC581-607
Acceso en línea:	https://doaj.org/article/93d9387f9b0442d697cdbaa8d8f1660e
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spelling	oai:doaj.org-article:93d9387f9b0442d697cdbaa8d8f1660e2021-11-12T19:57:15ZDevelopment and characterization of a unique anti‐IgE mouse monoclonal antibody cross‐reactive between human and canine IgE2050-452710.1002/iid3.531https://doaj.org/article/93d9387f9b0442d697cdbaa8d8f1660e2021-12-01T00:00:00Zhttps://doi.org/10.1002/iid3.531https://doaj.org/toc/2050-4527Abstract Background The efficacy assessment of human anti‐IgE monoclonal antibodies (mAbs) in animal models before clinical trials is hampered due to the lack of cross‐reactivity of anti‐IgE mAbs between species. Objective We developed CRE‐DR (an anti‐dog IgE monoclonal antibody), an anti‐IgE mouse mAb that recognizes canine and human IgE, and then examined its IgE specificity and cross‐reactivity between three animal and human species. Methods After mouse immunization with a synthetic peptide derived from canine IgE (282NTNDWIEGETYYC294), we generated a hybridoma producing CRE‐DR. The CRE‐DR purified from the ascites of hybridoma‐inoculated mice was used for ELISA and Western blot analysis to examine reactivity to dog, human, and rodent IgEs as well as recombinant bovine serum albumin (BSA)‐conjugated to canine, human, and rodent IgE amino acid peptides corresponding to the immunizing sequence. We then performed enzyme‐linked immunosorbent assays (ELISAs) for dog IgE using sera from dogs with atopic dermatitis (AD) after inhibition with canine IgE and IgG. The amino acid sequence recognized by CRE‐DR was identified by ELISA using synthetic peptides. Results CRE‐DR is a monoclonal mouse IgG1κ specific for dog IgE, and the ELISA values in atopic dog sera were inhibited by dog IgE, but not dog IgG. The binding of CRE‐DR to human IgE was relatively maintained, but not to rodent IgEs, which results were confirmed with the BSA‐conjugated IgE peptides of the various species. The CRE‐DR reactivity was supported by the comparison of amino acid sequence of CRE‐DR epitope, DWIEGETYYC, in dog IgE; one, two, and three amino acids were substituted in the human, rat, and mouse IgE epitopes, respectively. Conclusions and Clinical Relevance CRE‐DR is a mAb cross‐reactive to dog and human IgEs, which can allow the use of a dog model of allergy to test the efficacy of a CRE‐DR‐derived anti‐IgE therapeutic mAb before human clinical trials.Akiko KumagaiTakuya NaraMizuho UematsuYoko KakinumaTakashi SaitoKenichi MasudaWileyarticleanimalsepitopeshumansIgE \| monoclonal antibodyImmunologic diseases. AllergyRC581-607ENImmunity, Inflammation and Disease, Vol 9, Iss 4, Pp 1740-1748 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	animals epitopes humans IgE \| monoclonal antibody Immunologic diseases. Allergy RC581-607
spellingShingle	animals epitopes humans IgE \| monoclonal antibody Immunologic diseases. Allergy RC581-607 Akiko Kumagai Takuya Nara Mizuho Uematsu Yoko Kakinuma Takashi Saito Kenichi Masuda Development and characterization of a unique anti‐IgE mouse monoclonal antibody cross‐reactive between human and canine IgE
description	Abstract Background The efficacy assessment of human anti‐IgE monoclonal antibodies (mAbs) in animal models before clinical trials is hampered due to the lack of cross‐reactivity of anti‐IgE mAbs between species. Objective We developed CRE‐DR (an anti‐dog IgE monoclonal antibody), an anti‐IgE mouse mAb that recognizes canine and human IgE, and then examined its IgE specificity and cross‐reactivity between three animal and human species. Methods After mouse immunization with a synthetic peptide derived from canine IgE (282NTNDWIEGETYYC294), we generated a hybridoma producing CRE‐DR. The CRE‐DR purified from the ascites of hybridoma‐inoculated mice was used for ELISA and Western blot analysis to examine reactivity to dog, human, and rodent IgEs as well as recombinant bovine serum albumin (BSA)‐conjugated to canine, human, and rodent IgE amino acid peptides corresponding to the immunizing sequence. We then performed enzyme‐linked immunosorbent assays (ELISAs) for dog IgE using sera from dogs with atopic dermatitis (AD) after inhibition with canine IgE and IgG. The amino acid sequence recognized by CRE‐DR was identified by ELISA using synthetic peptides. Results CRE‐DR is a monoclonal mouse IgG1κ specific for dog IgE, and the ELISA values in atopic dog sera were inhibited by dog IgE, but not dog IgG. The binding of CRE‐DR to human IgE was relatively maintained, but not to rodent IgEs, which results were confirmed with the BSA‐conjugated IgE peptides of the various species. The CRE‐DR reactivity was supported by the comparison of amino acid sequence of CRE‐DR epitope, DWIEGETYYC, in dog IgE; one, two, and three amino acids were substituted in the human, rat, and mouse IgE epitopes, respectively. Conclusions and Clinical Relevance CRE‐DR is a mAb cross‐reactive to dog and human IgEs, which can allow the use of a dog model of allergy to test the efficacy of a CRE‐DR‐derived anti‐IgE therapeutic mAb before human clinical trials.
format	article
author	Akiko Kumagai Takuya Nara Mizuho Uematsu Yoko Kakinuma Takashi Saito Kenichi Masuda
author_facet	Akiko Kumagai Takuya Nara Mizuho Uematsu Yoko Kakinuma Takashi Saito Kenichi Masuda
author_sort	Akiko Kumagai
title	Development and characterization of a unique anti‐IgE mouse monoclonal antibody cross‐reactive between human and canine IgE
title_short	Development and characterization of a unique anti‐IgE mouse monoclonal antibody cross‐reactive between human and canine IgE
title_full	Development and characterization of a unique anti‐IgE mouse monoclonal antibody cross‐reactive between human and canine IgE
title_fullStr	Development and characterization of a unique anti‐IgE mouse monoclonal antibody cross‐reactive between human and canine IgE
title_full_unstemmed	Development and characterization of a unique anti‐IgE mouse monoclonal antibody cross‐reactive between human and canine IgE
title_sort	development and characterization of a unique anti‐ige mouse monoclonal antibody cross‐reactive between human and canine ige
publisher	Wiley
publishDate	2021
url	https://doaj.org/article/93d9387f9b0442d697cdbaa8d8f1660e
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_version_	1718430330387431424

Development and characterization of a unique anti‐IgE mouse monoclonal antibody cross‐reactive between human and canine IgE

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