Development of Antibody Detection ELISA Based on Immunoreactive Toxins and Toxin-Derived Peptides to Evaluate the Neutralization Potency of Equine Plasma against Naja atra in Taiwan

Naja atra, also known as Taiwanese cobra, is one of the most prevalent venomous snakes in Taiwan. Clinically, freeze-dried neurotoxic antivenom (FNAV) produced from horses by Taiwan Centers for Disease Control (CDC) has been the only approved treatment for N. atra</i&g...

Descripción completa

Guardado en:

Detalles Bibliográficos
Autores principales:	Chien-Chun Liu, Yung-Chin Hsiao, Lichieh Julie Chu, Po-Jung Wang, Chien-Hsin Liu, Wen-Chin Hsieh, Jau-Song Yu
Formato:	article
Lenguaje:	EN
Publicado:	MDPI AG 2021
Materias:	<i>Naja atra</i> freeze-dried neurotoxic antivenom neutralization potency neurotoxin epitope peptide-based ELISA Medicine R
Acceso en línea:	https://doaj.org/article/145d7e59cc4d4a6cb386ba47a3dff093
Etiquetas:	Agregar Etiqueta Sin Etiquetas, Sea el primero en etiquetar este registro!

id	oai:doaj.org-article:145d7e59cc4d4a6cb386ba47a3dff093
record_format	dspace
spelling	oai:doaj.org-article:145d7e59cc4d4a6cb386ba47a3dff0932021-11-25T19:09:06ZDevelopment of Antibody Detection ELISA Based on Immunoreactive Toxins and Toxin-Derived Peptides to Evaluate the Neutralization Potency of Equine Plasma against <i>Naja atra</i> in Taiwan10.3390/toxins131108182072-6651https://doaj.org/article/145d7e59cc4d4a6cb386ba47a3dff0932021-11-01T00:00:00Zhttps://www.mdpi.com/2072-6651/13/11/818https://doaj.org/toc/2072-6651<i>Naja atra</i>, also known as Taiwanese cobra, is one of the most prevalent venomous snakes in Taiwan. Clinically, freeze-dried neurotoxic antivenom (FNAV) produced from horses by Taiwan Centers for Disease Control (CDC) has been the only approved treatment for <i>N. atra</i> envenoming for the last few decades. During antivenom production, large numbers of mice are used in the in vivo assay to determine whether the neutralization potency of hyperimmunized equines is satisfactory for large-scale harvesting. However, this in vivo assay is extremely laborious, expensive, and significantly impairs animal welfare. In the present study, we aimed to develop an in vitro ELISA-based system that could serve as an alternative assay to evaluate the neutralization potency of plasma from hyperimmunized equines. We initially obtained 51 plasma samples with known (high or low) neutralization potency assessed in vivo from 9 hyperimmunized equines and subsequently determined their antibody titers against the five major protein components of <i>N. atra</i> venom (neurotoxin (NTX), phospholipase A2 (PLA<sub>2</sub>), cytotoxin (CTX), cysteine-rich secretory protein (CRISP), and snake venom metalloproteinase (SVMP)) via ELISA. The antibody titer against NTX was the most effective in discriminating between high and low potency plasma samples. To identify the specific epitope(s) of NTX recognized by neutralization potency-related antibodies, 17 consecutive NTX-derived pentadecapeptides were synthesized and used as antigens to probe the 51 equine plasma samples. Among the 17 peptides, immunoreactive signals for three consecutive peptides (NTX1-8, NTX1-9, and NTX1-10) were significantly higher in the high potency relative to low potency equine plasma groups (<i>p</i> < 0.0001). Our ELISA system based on NTX1-10 peptide (RWRDHRGYRTERGCG) encompassing residues 28–42 of NTX displayed optimal sensitivity (96.88%) and specificity (89.47%) for differentiating between high- and low-potency plasma samples (area under the receiver operating characteristic curve (AUC) = 0.95). The collective data clearly indicate that the antibody titer against NTX protein or derived peptides can be used to efficiently discriminate between high and low neutralization potency of plasma samples from venom-immunized horses. This newly developed antibody detection ELISA based on NTX or its peptide derivatives has good potential to complement or replace the in vivo rodent assay for determining whether the neutralization potency of equine plasma is satisfactory for large-scale harvesting in the antivenom production process against <i>N. atra.</i>Chien-Chun LiuYung-Chin HsiaoLichieh Julie ChuPo-Jung WangChien-Hsin LiuWen-Chin HsiehJau-Song YuMDPI AGarticle<i>Naja atra</i>freeze-dried neurotoxic antivenomneutralization potencyneurotoxinepitopepeptide-based ELISAMedicineRENToxins, Vol 13, Iss 818, p 818 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	<i>Naja atra</i> freeze-dried neurotoxic antivenom neutralization potency neurotoxin epitope peptide-based ELISA Medicine R
spellingShingle	<i>Naja atra</i> freeze-dried neurotoxic antivenom neutralization potency neurotoxin epitope peptide-based ELISA Medicine R Chien-Chun Liu Yung-Chin Hsiao Lichieh Julie Chu Po-Jung Wang Chien-Hsin Liu Wen-Chin Hsieh Jau-Song Yu Development of Antibody Detection ELISA Based on Immunoreactive Toxins and Toxin-Derived Peptides to Evaluate the Neutralization Potency of Equine Plasma against <i>Naja atra</i> in Taiwan
description	<i>Naja atra</i>, also known as Taiwanese cobra, is one of the most prevalent venomous snakes in Taiwan. Clinically, freeze-dried neurotoxic antivenom (FNAV) produced from horses by Taiwan Centers for Disease Control (CDC) has been the only approved treatment for <i>N. atra</i> envenoming for the last few decades. During antivenom production, large numbers of mice are used in the in vivo assay to determine whether the neutralization potency of hyperimmunized equines is satisfactory for large-scale harvesting. However, this in vivo assay is extremely laborious, expensive, and significantly impairs animal welfare. In the present study, we aimed to develop an in vitro ELISA-based system that could serve as an alternative assay to evaluate the neutralization potency of plasma from hyperimmunized equines. We initially obtained 51 plasma samples with known (high or low) neutralization potency assessed in vivo from 9 hyperimmunized equines and subsequently determined their antibody titers against the five major protein components of <i>N. atra</i> venom (neurotoxin (NTX), phospholipase A2 (PLA<sub>2</sub>), cytotoxin (CTX), cysteine-rich secretory protein (CRISP), and snake venom metalloproteinase (SVMP)) via ELISA. The antibody titer against NTX was the most effective in discriminating between high and low potency plasma samples. To identify the specific epitope(s) of NTX recognized by neutralization potency-related antibodies, 17 consecutive NTX-derived pentadecapeptides were synthesized and used as antigens to probe the 51 equine plasma samples. Among the 17 peptides, immunoreactive signals for three consecutive peptides (NTX1-8, NTX1-9, and NTX1-10) were significantly higher in the high potency relative to low potency equine plasma groups (<i>p</i> < 0.0001). Our ELISA system based on NTX1-10 peptide (RWRDHRGYRTERGCG) encompassing residues 28–42 of NTX displayed optimal sensitivity (96.88%) and specificity (89.47%) for differentiating between high- and low-potency plasma samples (area under the receiver operating characteristic curve (AUC) = 0.95). The collective data clearly indicate that the antibody titer against NTX protein or derived peptides can be used to efficiently discriminate between high and low neutralization potency of plasma samples from venom-immunized horses. This newly developed antibody detection ELISA based on NTX or its peptide derivatives has good potential to complement or replace the in vivo rodent assay for determining whether the neutralization potency of equine plasma is satisfactory for large-scale harvesting in the antivenom production process against <i>N. atra.</i>
format	article
author	Chien-Chun Liu Yung-Chin Hsiao Lichieh Julie Chu Po-Jung Wang Chien-Hsin Liu Wen-Chin Hsieh Jau-Song Yu
author_facet	Chien-Chun Liu Yung-Chin Hsiao Lichieh Julie Chu Po-Jung Wang Chien-Hsin Liu Wen-Chin Hsieh Jau-Song Yu
author_sort	Chien-Chun Liu
title	Development of Antibody Detection ELISA Based on Immunoreactive Toxins and Toxin-Derived Peptides to Evaluate the Neutralization Potency of Equine Plasma against <i>Naja atra</i> in Taiwan
title_short	Development of Antibody Detection ELISA Based on Immunoreactive Toxins and Toxin-Derived Peptides to Evaluate the Neutralization Potency of Equine Plasma against <i>Naja atra</i> in Taiwan
title_full	Development of Antibody Detection ELISA Based on Immunoreactive Toxins and Toxin-Derived Peptides to Evaluate the Neutralization Potency of Equine Plasma against <i>Naja atra</i> in Taiwan
title_fullStr	Development of Antibody Detection ELISA Based on Immunoreactive Toxins and Toxin-Derived Peptides to Evaluate the Neutralization Potency of Equine Plasma against <i>Naja atra</i> in Taiwan
title_full_unstemmed	Development of Antibody Detection ELISA Based on Immunoreactive Toxins and Toxin-Derived Peptides to Evaluate the Neutralization Potency of Equine Plasma against <i>Naja atra</i> in Taiwan
title_sort	development of antibody detection elisa based on immunoreactive toxins and toxin-derived peptides to evaluate the neutralization potency of equine plasma against <i>naja atra</i> in taiwan
publisher	MDPI AG
publishDate	2021
url	https://doaj.org/article/145d7e59cc4d4a6cb386ba47a3dff093
work_keys_str_mv	AT chienchunliu developmentofantibodydetectionelisabasedonimmunoreactivetoxinsandtoxinderivedpeptidestoevaluatetheneutralizationpotencyofequineplasmaagainstinajaatraiintaiwan AT yungchinhsiao developmentofantibodydetectionelisabasedonimmunoreactivetoxinsandtoxinderivedpeptidestoevaluatetheneutralizationpotencyofequineplasmaagainstinajaatraiintaiwan AT lichiehjuliechu developmentofantibodydetectionelisabasedonimmunoreactivetoxinsandtoxinderivedpeptidestoevaluatetheneutralizationpotencyofequineplasmaagainstinajaatraiintaiwan AT pojungwang developmentofantibodydetectionelisabasedonimmunoreactivetoxinsandtoxinderivedpeptidestoevaluatetheneutralizationpotencyofequineplasmaagainstinajaatraiintaiwan AT chienhsinliu developmentofantibodydetectionelisabasedonimmunoreactivetoxinsandtoxinderivedpeptidestoevaluatetheneutralizationpotencyofequineplasmaagainstinajaatraiintaiwan AT wenchinhsieh developmentofantibodydetectionelisabasedonimmunoreactivetoxinsandtoxinderivedpeptidestoevaluatetheneutralizationpotencyofequineplasmaagainstinajaatraiintaiwan AT jausongyu developmentofantibodydetectionelisabasedonimmunoreactivetoxinsandtoxinderivedpeptidestoevaluatetheneutralizationpotencyofequineplasmaagainstinajaatraiintaiwan
_version_	1718410199022174208

Development of Antibody Detection ELISA Based on Immunoreactive Toxins and Toxin-Derived Peptides to Evaluate the Neutralization Potency of Equine Plasma against <i>Naja atra</i> in Taiwan

Ejemplares similares