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Sumateran wild boar (Sus scrofa vittatus) meat antibody production as immunodiagnostic reagent candidate
AIM: Meat authentication gives significance values in view of religious, food safety, public health, quality assurance, and legal concern. Most of the meat authentication is based on molecular assay; a simpler method to authenticate meat is needed to develop. An immunoassays technique may offer a so...
Autores principales: | , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Veterinary World
2019
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6515818/ https://www.ncbi.nlm.nih.gov/pubmed/31190701 http://dx.doi.org/10.14202/vetworld.2019.477-482 |
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author | Adiningsih, Melani Wahyu Soejoedono, Retno Damajanti Adji, Rahmat Setya Putri, Dwi Desmiyeni Purnawarman, Trioso Latif, Hadri Poetri, Okti Nadia |
author_facet | Adiningsih, Melani Wahyu Soejoedono, Retno Damajanti Adji, Rahmat Setya Putri, Dwi Desmiyeni Purnawarman, Trioso Latif, Hadri Poetri, Okti Nadia |
author_sort | Adiningsih, Melani Wahyu |
collection | PubMed |
description | AIM: Meat authentication gives significance values in view of religious, food safety, public health, quality assurance, and legal concern. Most of the meat authentication is based on molecular assay; a simpler method to authenticate meat is needed to develop. An immunoassays technique may offer a solution for simpler test. The aim of our current study was to develop a polyclonal antibody of Sus scrofa vittatus (Sumateran wild boar) as an immunodiagnostic reagent candidate. MATERIALS AND METHODS: Three male New Zealand white rabbits were used in this study for antibody production. Antigen used was meat extract of Sumateran wild boar, each rabbit was immunized with meat extract antigen (0.5 mg/ml) emulsified in Freund’s complete adjuvant at a 1:1 (v/v) ratio as much as 1 ml at subcutaneous route. Booster was carried out 3 times with interval time of 14 days, using meat extract antigen emulsified in Freund’s incomplete adjuvant at a 1:1 (v/v) ratio. Serum samples were taken every week, start from 1 week after the first immunization up to 1 week after the third booster. Antibody purification was performed using ammonium sulfate precipitation and Protein A. The presence of specific antibody was determined using agar gel precipitation test and enzyme-linked immunosorbent assay, while purified specific IgG was characterized using sodium dodecyl sulfate-polyacrylamide gel electrophoresis method. RESULTS: Specific antibody was detected at 14 days after the first immunization and still detected until 2 weeks after the third booster. Highest absorbance of specific antibody was detected 1 week after the third booster. CONCLUSION: The present study demonstrated that specific antibody of Sumateran wild boar is favorable to be produced in rabbit and showed that antibody produced is applicable to detect Sumateran wild boar meat antigen in immunodiffusion assay, indicating that it is promising as a reagent candidate in immunodiagnostic assay/kit. |
format | Online Article Text |
id | pubmed-6515818 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2019 |
publisher | Veterinary World |
record_format | MEDLINE/PubMed |
spelling | pubmed-65158182019-06-12 Sumateran wild boar (Sus scrofa vittatus) meat antibody production as immunodiagnostic reagent candidate Adiningsih, Melani Wahyu Soejoedono, Retno Damajanti Adji, Rahmat Setya Putri, Dwi Desmiyeni Purnawarman, Trioso Latif, Hadri Poetri, Okti Nadia Vet World Research Article AIM: Meat authentication gives significance values in view of religious, food safety, public health, quality assurance, and legal concern. Most of the meat authentication is based on molecular assay; a simpler method to authenticate meat is needed to develop. An immunoassays technique may offer a solution for simpler test. The aim of our current study was to develop a polyclonal antibody of Sus scrofa vittatus (Sumateran wild boar) as an immunodiagnostic reagent candidate. MATERIALS AND METHODS: Three male New Zealand white rabbits were used in this study for antibody production. Antigen used was meat extract of Sumateran wild boar, each rabbit was immunized with meat extract antigen (0.5 mg/ml) emulsified in Freund’s complete adjuvant at a 1:1 (v/v) ratio as much as 1 ml at subcutaneous route. Booster was carried out 3 times with interval time of 14 days, using meat extract antigen emulsified in Freund’s incomplete adjuvant at a 1:1 (v/v) ratio. Serum samples were taken every week, start from 1 week after the first immunization up to 1 week after the third booster. Antibody purification was performed using ammonium sulfate precipitation and Protein A. The presence of specific antibody was determined using agar gel precipitation test and enzyme-linked immunosorbent assay, while purified specific IgG was characterized using sodium dodecyl sulfate-polyacrylamide gel electrophoresis method. RESULTS: Specific antibody was detected at 14 days after the first immunization and still detected until 2 weeks after the third booster. Highest absorbance of specific antibody was detected 1 week after the third booster. CONCLUSION: The present study demonstrated that specific antibody of Sumateran wild boar is favorable to be produced in rabbit and showed that antibody produced is applicable to detect Sumateran wild boar meat antigen in immunodiffusion assay, indicating that it is promising as a reagent candidate in immunodiagnostic assay/kit. Veterinary World 2019 2019-04-01 /pmc/articles/PMC6515818/ /pubmed/31190701 http://dx.doi.org/10.14202/vetworld.2019.477-482 Text en Copyright: © Adiningsih, et al. http://creativecommons.org/licenses/by/4.0 Open Access. This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated. |
spellingShingle | Research Article Adiningsih, Melani Wahyu Soejoedono, Retno Damajanti Adji, Rahmat Setya Putri, Dwi Desmiyeni Purnawarman, Trioso Latif, Hadri Poetri, Okti Nadia Sumateran wild boar (Sus scrofa vittatus) meat antibody production as immunodiagnostic reagent candidate |
title | Sumateran wild boar (Sus scrofa vittatus) meat antibody production as immunodiagnostic reagent candidate |
title_full | Sumateran wild boar (Sus scrofa vittatus) meat antibody production as immunodiagnostic reagent candidate |
title_fullStr | Sumateran wild boar (Sus scrofa vittatus) meat antibody production as immunodiagnostic reagent candidate |
title_full_unstemmed | Sumateran wild boar (Sus scrofa vittatus) meat antibody production as immunodiagnostic reagent candidate |
title_short | Sumateran wild boar (Sus scrofa vittatus) meat antibody production as immunodiagnostic reagent candidate |
title_sort | sumateran wild boar (sus scrofa vittatus) meat antibody production as immunodiagnostic reagent candidate |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6515818/ https://www.ncbi.nlm.nih.gov/pubmed/31190701 http://dx.doi.org/10.14202/vetworld.2019.477-482 |
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