Development of an Immunoassay for the Detection of Amyloid Beta 1-42 and Its Application in Urine Samples
Amyloid beta peptides (Aβ1-42) have been found to be associated with the cause of Alzheimer’s disease (AD) and dementia. Currently, methods for detecting Aβ1-42 are complicated and expensive. The present study is aimed at developing an indirect competitive enzyme-linked immunosorbent assay (ic-ELISA...
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| Format: | Article |
| Language: | English |
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Wiley
2020-01-01
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| Series: | Journal of Immunology Research |
| Online Access: | http://dx.doi.org/10.1155/2020/8821181 |
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| author | Anurak Wongta Surat Hongsibsong Somporn Chantara Mookda Pattarawarapan Ratana Sapbamrer Korawan Sringarm Zhen-Lin Xu Hong Wang |
| author_facet | Anurak Wongta Surat Hongsibsong Somporn Chantara Mookda Pattarawarapan Ratana Sapbamrer Korawan Sringarm Zhen-Lin Xu Hong Wang |
| author_sort | Anurak Wongta |
| collection | DOAJ |
| description | Amyloid beta peptides (Aβ1-42) have been found to be associated with the cause of Alzheimer’s disease (AD) and dementia. Currently, methods for detecting Aβ1-42 are complicated and expensive. The present study is aimed at developing an indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) to detect Aβ1-42 by using a polyclonal antibody from alpaca, an application used in urine samples. The serum was collected from the alpaca after immunizing it with Aβ1-42 at 500 μg/injection 5 times. The ic-ELISA was developed and showed a half-maximal inhibitory concentration (IC50) of 103.20 ng/ml. The limit of detection (LOD) was 0.39 ng/100 μl. The cross-reactivity was tested with Aβ1-40 and 8 synthesized peptides that had sequence similarities to parts of Aβ1-42. The cross-reactivity of Aβ1-40 and peptide 1 (DAEFRHDSGYE) was 55% and 69.4%, respectively. The ic-ELISA was applied to analyze Aβ1-42 in the urine and precipitated protein urine samples. This method can be used for detecting a normal level of total soluble Aβ (approximately 1 ng in 5 mg of precipitated urine protein) and can be used for detecting the early stages of AD. It is considered to be an easy and inexpensive method for monitoring and diagnosing AD. |
| format | Article |
| id | doaj-art-d3f9087f65d44f2d9374b4248b251f81 |
| institution | DOAJ |
| issn | 2314-8861 2314-7156 |
| language | English |
| publishDate | 2020-01-01 |
| publisher | Wiley |
| record_format | Article |
| series | Journal of Immunology Research |
| spelling | doaj-art-d3f9087f65d44f2d9374b4248b251f812025-08-20T03:23:47ZengWileyJournal of Immunology Research2314-88612314-71562020-01-01202010.1155/2020/88211818821181Development of an Immunoassay for the Detection of Amyloid Beta 1-42 and Its Application in Urine SamplesAnurak Wongta0Surat Hongsibsong1Somporn Chantara2Mookda Pattarawarapan3Ratana Sapbamrer4Korawan Sringarm5Zhen-Lin Xu6Hong Wang7Environmental Science Ph.D. Program, Faculty of Science, Chiang Mai University, Chiang Mai 50200, ThailandEnvironmental Science Ph.D. Program, Faculty of Science, Chiang Mai University, Chiang Mai 50200, ThailandEnvironmental Chemistry Research Laboratory, Department of Chemistry, Faculty of Science, Chiang Mai University, Chiang Mai 50200, ThailandDepartment of Chemistry and Center of Excellence for Innovation in Chemistry, Faculty of Science, Chiang Mai University, Chiang Mai 50200, ThailandDepartment of Community Medicine, Faculty of Medicine, Chiang Mai University, Chiang Mai 50200, ThailandCluster of Research and Development of Pharmaceutical and Natural Products Innovation for Human or Animal, Chiang Mai University, Chiang Mai 50200, ThailandGuangdong Provincial Key Laboratory of Food Quality and Safety, College of Food Science, South China Agricultural University, Guangzhou 510642, ChinaGuangdong Provincial Key Laboratory of Food Quality and Safety, College of Food Science, South China Agricultural University, Guangzhou 510642, ChinaAmyloid beta peptides (Aβ1-42) have been found to be associated with the cause of Alzheimer’s disease (AD) and dementia. Currently, methods for detecting Aβ1-42 are complicated and expensive. The present study is aimed at developing an indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) to detect Aβ1-42 by using a polyclonal antibody from alpaca, an application used in urine samples. The serum was collected from the alpaca after immunizing it with Aβ1-42 at 500 μg/injection 5 times. The ic-ELISA was developed and showed a half-maximal inhibitory concentration (IC50) of 103.20 ng/ml. The limit of detection (LOD) was 0.39 ng/100 μl. The cross-reactivity was tested with Aβ1-40 and 8 synthesized peptides that had sequence similarities to parts of Aβ1-42. The cross-reactivity of Aβ1-40 and peptide 1 (DAEFRHDSGYE) was 55% and 69.4%, respectively. The ic-ELISA was applied to analyze Aβ1-42 in the urine and precipitated protein urine samples. This method can be used for detecting a normal level of total soluble Aβ (approximately 1 ng in 5 mg of precipitated urine protein) and can be used for detecting the early stages of AD. It is considered to be an easy and inexpensive method for monitoring and diagnosing AD.http://dx.doi.org/10.1155/2020/8821181 |
| spellingShingle | Anurak Wongta Surat Hongsibsong Somporn Chantara Mookda Pattarawarapan Ratana Sapbamrer Korawan Sringarm Zhen-Lin Xu Hong Wang Development of an Immunoassay for the Detection of Amyloid Beta 1-42 and Its Application in Urine Samples Journal of Immunology Research |
| title | Development of an Immunoassay for the Detection of Amyloid Beta 1-42 and Its Application in Urine Samples |
| title_full | Development of an Immunoassay for the Detection of Amyloid Beta 1-42 and Its Application in Urine Samples |
| title_fullStr | Development of an Immunoassay for the Detection of Amyloid Beta 1-42 and Its Application in Urine Samples |
| title_full_unstemmed | Development of an Immunoassay for the Detection of Amyloid Beta 1-42 and Its Application in Urine Samples |
| title_short | Development of an Immunoassay for the Detection of Amyloid Beta 1-42 and Its Application in Urine Samples |
| title_sort | development of an immunoassay for the detection of amyloid beta 1 42 and its application in urine samples |
| url | http://dx.doi.org/10.1155/2020/8821181 |
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