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Isolation and Production of Human Monoclonal Antibody Proteins against a Toxocara canis Excretory–Secretory Recombinant Antigen

Toxocariasis is a widespread zoonotic parasitic disease with a significant socioeconomic impact, particularly on underprivileged communities. Limitations of existing diagnostic tools and vague presenting symptoms may lead to misdiagnosis, thus underestimating the actual global impact of the disease....

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Autores principales: Baharudeen, Zamrina, Noordin, Rahmah, Soon, Lim Theam, Balachandra, Dinesh, Anuar, Nor Suhada, Mustafa, Fatin Hamimi, Rahumatullah, Anizah
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9698333/
https://www.ncbi.nlm.nih.gov/pubmed/36364983
http://dx.doi.org/10.3390/pathogens11111232
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author Baharudeen, Zamrina
Noordin, Rahmah
Soon, Lim Theam
Balachandra, Dinesh
Anuar, Nor Suhada
Mustafa, Fatin Hamimi
Rahumatullah, Anizah
author_facet Baharudeen, Zamrina
Noordin, Rahmah
Soon, Lim Theam
Balachandra, Dinesh
Anuar, Nor Suhada
Mustafa, Fatin Hamimi
Rahumatullah, Anizah
author_sort Baharudeen, Zamrina
collection PubMed
description Toxocariasis is a widespread zoonotic parasitic disease with a significant socioeconomic impact, particularly on underprivileged communities. Limitations of existing diagnostic tools and vague presenting symptoms may lead to misdiagnosis, thus underestimating the actual global impact of the disease. The present study describes the isolation and production of novel recombinant monoclonal antibodies against Toxocara canis recombinant TES-26 antigen (rTES-26) utilizing a human helminth scFv phage display library. The isolated antibody clones were characterized based on their gene sequences and binding characteristics. Three clones representing unique gene families (clone 48: IgHV3-LV1; clone 49: IgHV3-LV3; clone 50: IgHV6-LV3) were isolated, but only clones 48 and 49 showed successful insertion of the full-length scFv antibody sequence after sub-cloning. Both clones produced antibody proteins of good solubility and satisfactory yield and purity. Binding assays via Western blot and ELISA using rTES-26 and Toxocara canis native protein showed that both monoclonal antibodies were highly specific and sensitive to the target antigen. A preliminary antigen detection ELISA showed the diagnostic potential of the monoclonal antibody proteins. The proteins can also be useful in studying host–parasite interactions and therapeutic applications.
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spelling pubmed-96983332022-11-26 Isolation and Production of Human Monoclonal Antibody Proteins against a Toxocara canis Excretory–Secretory Recombinant Antigen Baharudeen, Zamrina Noordin, Rahmah Soon, Lim Theam Balachandra, Dinesh Anuar, Nor Suhada Mustafa, Fatin Hamimi Rahumatullah, Anizah Pathogens Article Toxocariasis is a widespread zoonotic parasitic disease with a significant socioeconomic impact, particularly on underprivileged communities. Limitations of existing diagnostic tools and vague presenting symptoms may lead to misdiagnosis, thus underestimating the actual global impact of the disease. The present study describes the isolation and production of novel recombinant monoclonal antibodies against Toxocara canis recombinant TES-26 antigen (rTES-26) utilizing a human helminth scFv phage display library. The isolated antibody clones were characterized based on their gene sequences and binding characteristics. Three clones representing unique gene families (clone 48: IgHV3-LV1; clone 49: IgHV3-LV3; clone 50: IgHV6-LV3) were isolated, but only clones 48 and 49 showed successful insertion of the full-length scFv antibody sequence after sub-cloning. Both clones produced antibody proteins of good solubility and satisfactory yield and purity. Binding assays via Western blot and ELISA using rTES-26 and Toxocara canis native protein showed that both monoclonal antibodies were highly specific and sensitive to the target antigen. A preliminary antigen detection ELISA showed the diagnostic potential of the monoclonal antibody proteins. The proteins can also be useful in studying host–parasite interactions and therapeutic applications. MDPI 2022-10-25 /pmc/articles/PMC9698333/ /pubmed/36364983 http://dx.doi.org/10.3390/pathogens11111232 Text en © 2022 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Baharudeen, Zamrina
Noordin, Rahmah
Soon, Lim Theam
Balachandra, Dinesh
Anuar, Nor Suhada
Mustafa, Fatin Hamimi
Rahumatullah, Anizah
Isolation and Production of Human Monoclonal Antibody Proteins against a Toxocara canis Excretory–Secretory Recombinant Antigen
title Isolation and Production of Human Monoclonal Antibody Proteins against a Toxocara canis Excretory–Secretory Recombinant Antigen
title_full Isolation and Production of Human Monoclonal Antibody Proteins against a Toxocara canis Excretory–Secretory Recombinant Antigen
title_fullStr Isolation and Production of Human Monoclonal Antibody Proteins against a Toxocara canis Excretory–Secretory Recombinant Antigen
title_full_unstemmed Isolation and Production of Human Monoclonal Antibody Proteins against a Toxocara canis Excretory–Secretory Recombinant Antigen
title_short Isolation and Production of Human Monoclonal Antibody Proteins against a Toxocara canis Excretory–Secretory Recombinant Antigen
title_sort isolation and production of human monoclonal antibody proteins against a toxocara canis excretory–secretory recombinant antigen
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9698333/
https://www.ncbi.nlm.nih.gov/pubmed/36364983
http://dx.doi.org/10.3390/pathogens11111232
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