Evaluation of antidiphtheria toxin nanobodies

Ghada H ShakerDepartment of Microbiology and Immunology, Faculty of Pharmacy, King Saud University, Riyadh, Kingdom of Saudi ArabiaAbstract: Nanobodies are the smallest fragments of naturally occurring single-domain antibodies that have evolved to be fully functional in the absence of a light chain....

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Autor principal:	Ghada H Shaker
Formato:	article
Lenguaje:	EN
Publicado:	Dove Medical Press 2010
Materias:	Medical technology R855-855.5 Chemical technology TP1-1185
Acceso en línea:	https://doaj.org/article/eb1168f5a0474e2c8f8a2a382d701d48
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spelling	oai:doaj.org-article:eb1168f5a0474e2c8f8a2a382d701d482021-12-02T06:12:10ZEvaluation of antidiphtheria toxin nanobodies1177-8903https://doaj.org/article/eb1168f5a0474e2c8f8a2a382d701d482010-06-01T00:00:00Zhttp://www.dovepress.com/evaluation-of-antidiphtheria-toxin-nanobodies-a4608https://doaj.org/toc/1177-8903Ghada H ShakerDepartment of Microbiology and Immunology, Faculty of Pharmacy, King Saud University, Riyadh, Kingdom of Saudi ArabiaAbstract: Nanobodies are the smallest fragments of naturally occurring single-domain antibodies that have evolved to be fully functional in the absence of a light chain. Conventional antibodies are glycoproteins comprising two heavy and two light chains. Surprisingly, all members of the Camelidae family possess a fraction of antibodies devoid of both light chains and the first constant domain. These types of antibodies are known as heavy-chain antibody (HcAb) nanobodies. There are three subclasses of IgG in dromedaries, namely IgG1, IgG2, and IgG3 of which IgG2 and IgG3 are of the HcAb type. These heavy chain antibodies constitute approximately 50% of the IgG in llama serum and as much as 75% of the IgG in camel serum. In the present work, the different IgG subclasses from an immunized camel (Camelus dromedarius) with divalent diphtheria-tetanus vaccine were purified using their different affinity for protein A and protein G and their absorbance measured at 280 nm. Purity control and characterization by 12% sodium dodecyl sulfate-polyacrylamide gel electrophoresis of IgG subclasses was done under reducing conditions. Protein bands were visualized after staining with Coomassie Blue, showing two bands at 50 kDa and 30 kDa for IgG1, while IgG2 and IgG3 produced only one band at 46 kDa and 43 kDa, respectively. An enzyme-linked immunosorbent assay test using diphtheria toxin and purified IgG subclasses from the immunized camel were performed to evaluate their efficiency. Compared with conventional IgG1, heavy chain antibodies (nanobodies) were shown to be more efficient in binding to diphtheria toxin antigen. This study revealed the possibility of using IgG2 and IgG3 nanobodies as an effective antitoxin for the treatment of diphtheria in humans.Keywords: camel, heavy chain antibody, HcAb, nanobodies, immunoglobulin, IgG subclasses, diphtheria toxin Ghada H ShakerDove Medical PressarticleMedical technologyR855-855.5Chemical technologyTP1-1185ENNanotechnology, Science and Applications, Vol 2010, Iss default, Pp 29-35 (2010)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medical technology R855-855.5 Chemical technology TP1-1185
spellingShingle	Medical technology R855-855.5 Chemical technology TP1-1185 Ghada H Shaker Evaluation of antidiphtheria toxin nanobodies
description	Ghada H ShakerDepartment of Microbiology and Immunology, Faculty of Pharmacy, King Saud University, Riyadh, Kingdom of Saudi ArabiaAbstract: Nanobodies are the smallest fragments of naturally occurring single-domain antibodies that have evolved to be fully functional in the absence of a light chain. Conventional antibodies are glycoproteins comprising two heavy and two light chains. Surprisingly, all members of the Camelidae family possess a fraction of antibodies devoid of both light chains and the first constant domain. These types of antibodies are known as heavy-chain antibody (HcAb) nanobodies. There are three subclasses of IgG in dromedaries, namely IgG1, IgG2, and IgG3 of which IgG2 and IgG3 are of the HcAb type. These heavy chain antibodies constitute approximately 50% of the IgG in llama serum and as much as 75% of the IgG in camel serum. In the present work, the different IgG subclasses from an immunized camel (Camelus dromedarius) with divalent diphtheria-tetanus vaccine were purified using their different affinity for protein A and protein G and their absorbance measured at 280 nm. Purity control and characterization by 12% sodium dodecyl sulfate-polyacrylamide gel electrophoresis of IgG subclasses was done under reducing conditions. Protein bands were visualized after staining with Coomassie Blue, showing two bands at 50 kDa and 30 kDa for IgG1, while IgG2 and IgG3 produced only one band at 46 kDa and 43 kDa, respectively. An enzyme-linked immunosorbent assay test using diphtheria toxin and purified IgG subclasses from the immunized camel were performed to evaluate their efficiency. Compared with conventional IgG1, heavy chain antibodies (nanobodies) were shown to be more efficient in binding to diphtheria toxin antigen. This study revealed the possibility of using IgG2 and IgG3 nanobodies as an effective antitoxin for the treatment of diphtheria in humans.Keywords: camel, heavy chain antibody, HcAb, nanobodies, immunoglobulin, IgG subclasses, diphtheria toxin
format	article
author	Ghada H Shaker
author_facet	Ghada H Shaker
author_sort	Ghada H Shaker
title	Evaluation of antidiphtheria toxin nanobodies
title_short	Evaluation of antidiphtheria toxin nanobodies
title_full	Evaluation of antidiphtheria toxin nanobodies
title_fullStr	Evaluation of antidiphtheria toxin nanobodies
title_full_unstemmed	Evaluation of antidiphtheria toxin nanobodies
title_sort	evaluation of antidiphtheria toxin nanobodies
publisher	Dove Medical Press
publishDate	2010
url	https://doaj.org/article/eb1168f5a0474e2c8f8a2a382d701d48
work_keys_str_mv	AT ghadahshaker evaluationofantidiphtheriatoxinnanobodies
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Evaluation of antidiphtheria toxin nanobodies

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