Preparation of Polyclonal Antibody Against Domoic Acid and Establishment of Indirect Competitive ELISA Assay

LI Bingxi; LIU Jing; LI Yang; YE Jianmin

doi:10.6054/j.jscnun.2020029

Journal of South China Normal University (Natural Science Edition) > 2020 > 52(2): 68-76. > DOI: 10.6054/j.jscnun.2020029

LI Bingxi, LIU Jing, LI Yang, YE Jianmin. Preparation of Polyclonal Antibody Against Domoic Acid and Establishment of Indirect Competitive ELISA Assay[J]. Journal of South China Normal University (Natural Science Edition), 2020, 52(2): 68-76. DOI: 10.6054/j.jscnun.2020029

Citation:

PDF (1198 KB)

Preparation of Polyclonal Antibody Against Domoic Acid and Establishment of Indirect Competitive ELISA Assay

School of Life Sciences, South China Normal University//Guangzhou Guangdong Provincial Key Laboratory for Healthy and Safe Aquaculture, Guangzhou 510631, China

More Information

Received Date: January 20, 2019
Available Online: March 21, 2021

Graphical Abstract

Abstract

Abstract

Domoic acid (DA), the main component of amnesic shellfish poisoning (ASP), can be bio-concentrated in fish and shellfish and have toxic effects on humans through the transmission of food chains. DA, a small molecule hapten, was coupled to the carrier protein keyhole limpet hemocyanin (KLH) and bovine serum albumin (BSA) with the activation ester method to prepare complete immunogen and coating antigen. DA-KLH and DA-BSA were successfully conjugated using 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC) and N-hydroxysuccinimide (NHS) and their conjugating ratios were 44 : 1 and 16 : 1, respectively, according to analyses with UV-visible spectrophotometry and SDS polypropylene gel electrophoresis (SDS-PAGE). The DA-specific titer of antiserum of 210 000 units/mL was obtained after immunizing the mice four times with DA-KLH. The optimal working concentrations and conditions were determined for detecting DA with the indirect competitive ELISA (ic-ELISA), and the DA detection standard curve was successfully plotted in the concentration range of 100 ng/mL~10 μg/mL. The rapid quantitative detection of DA with the established indirect competitive ELISA method might provide guidance for the development of the first-line seafood algae toxin detection kit and the red tide toxin monitoring system.
- amnesia shellfish poisoning,
- domoic acid,
- polyclonal antibody,
- ic-ELISA

FullText(HTML)

References (34)

References

[1]	徐吟梅. 2010年我国海洋环境质量总体较好[J].现代渔业信息, 2011(6):30-31. http://www.cqvip.com/QK/90549X/201106/38264405.html
[2]	张青田.中国海域赤潮发生趋势的年际变化[J].中国环境监测, 2013, 29(5):98-102. doi: 10.3969/j.issn.1002-6002.2013.05.019 ZHANG Q T. Reviewon the annual variation of red tides in China sea[J]. Environmental Monitoring in China, 2013, 29(5):98-102. doi: 10.3969/j.issn.1002-6002.2013.05.019
[3]	李士虎, 吴建新, 李庭古, 等.赤潮的危害、成因及对策[J].水利渔业, 2003, 3(6):38-39. doi: 10.3969/j.issn.1003-1278.2003.06.019 LI S H, WU J X, LI T G, et al. Harm, causes and countermeasures of red tide[J]. Reservoir Fisheries, 2003, 23(6):38-39. doi: 10.3969/j.issn.1003-1278.2003.06.019
[4]	阮雯, 纪炜炜, 岳冬冬, 等.贝类生物毒素研究进展[J].渔业信息与战略, 2017, 32(4):276-280. http://d.old.wanfangdata.com.cn/Periodical/xdyyxx201704006 RUAN W, JI W W, YUE D D, et al. Research advances in shellfish biotoxins[J]. Fishery Information and Strategy, 2017(4):276-280. http://d.old.wanfangdata.com.cn/Periodical/xdyyxx201704006
[5]	MRACI S C P. Shellfish poisoning: public health risk, quality assurance and analytical detection[J]. Chemistry in Australia, 1995, 62(2):22-28.
[6]	刘永健, 刘娜, 刘仁沿, 等.赤潮毒素研究进展[J].海洋环境科学, 2008, 27(S2):151-159. http://www.wanfangdata.com.cn/details/detail.do?_type=perio&id=hyhjkx2008z2032 LIU Y J, LIU N, LIU Y R, et al. Progress on red tide toxins[J]. Marine Environmental Science, 2008, 27(S2):151-159. http://www.wanfangdata.com.cn/details/detail.do?_type=perio&id=hyhjkx2008z2032
[7]	于仁成, 罗璇.我国近海有毒藻和藻毒素的研究现状与展望[J].海洋科学集刊, 2016(1):155-166. http://d.old.wanfangdata.com.cn/Periodical/hykjjk201601011 YU R C, LUO X. Status and researchperpestives on toxic algae and phycotoxins in the coastal waters of China[J]. Studia Marina Sinica, 2016(1):155-166. http://d.old.wanfangdata.com.cn/Periodical/hykjjk201601011
[8]	LELONG A, HEGARET H, SOUDANT P, et al.Pseudo-nitzschia (Bacillariophyceae) species, domoic acid and amnesic shellfish poisoning: revisiting previous paradigms[J]. Phycologia, 2012, 51(2):168-216. doi: 10.2216/11-37.1
[9]	高虹, 陈西平.软骨藻酸的毒性作用机制研究概况[J].环境卫生学杂志, 2002, 29(5):297-299. http://d.old.wanfangdata.com.cn/Periodical/gwyx-wsx200205009 GAO H, CHEN X P. Survey of the mechanism of toxic mechanism of domoic acid[J]. Journal of Environmental Hygiene, 2002, 29(5):297-299. http://d.old.wanfangdata.com.cn/Periodical/gwyx-wsx200205009
[10]	陈西平, 王成斌.软骨藻酸和谷氨酸受体[J].卫生研究, 2000, 29(4):255-256. doi: 10.3969/j.issn.1000-8020.2000.04.024 CHN X P, WANG C B. Domoic acid andgiutamate recertors[J]. Journal of Hygiene Research, 2000, 29(4):255-256. doi: 10.3969/j.issn.1000-8020.2000.04.024
[11]	WRIGHT J L C, BOYD R K, FREITAS A S W, et al. Identification of domoic acid, a neuroexcitatory amino acid, in toxic mussels from eastern Prince Edward Island[J]. Canadian Journal of Chemistry, 1989, 67(3):481-490. doi: 10.1139/v89-075
[12]	DICKEY R W, FRYXELL G A, GRANADE H R, et al. Detection of the marine toxins okadaic acid and domoic acid in shellfish and phytoplankton in the Gulf of Mexico[J]. Toxicon Official Journal of the International Society on Toxinology, 1992, 30(3):355-359. doi: 10.1016/0041-0101(92)90877-8
[13]	NOGUCHI T, ARAKAWA O. Distribution of domoic acid in seaweeds occurring inkagoshima, southern Japan[J]. Advances in Experimental Medicine and Biology, 1996, 391:521-526. http://www.ncbi.nlm.nih.gov/pubmed/8726088
[14]	SCHOLIN C A, GULLAND F, DOUCETTE G J, et al. Mortality of sea lions along the central California coast linked to a toxic diatom bloom[J]. Nature, 2000, 403(6765):80-84. doi: 10.1038/47481
[15]	杨平, 林怡芳.海洋生物毒素研究现状[J].环境卫生学杂志, 1997 (2):112-115. http://www.cnki.com.cn/Article/CJFDTotal-GWYX702.012.htm YANG P, LIN Y F. Advances in detection methods of red tide toxins[J]. Journal of Environmental Hygiene, 1997(2):112-115. http://www.cnki.com.cn/Article/CJFDTotal-GWYX702.012.htm
[16]	GOLDSTEIN T, ZABKA T S, DELONG R L, et al. The role of domoic acid in abortion and premature parturition of California sea lions (Zalophus californianus) on San Miguel Island, California[J]. Journal of Wildlife Diseases, 2009, 45(1):91-108. doi: 10.7589/0090-3558-45.1.91
[17]	PULIDO O M. Domoic acid toxicologic pathology: a review[J]. Marine Drugs, 2008, 6(2):180-219. http://d.old.wanfangdata.com.cn/OAPaper/oai_doaj-articles_287ec623bad25afaeed6c080744ba1ca
[18]	吕会田, 乐加昌, 陈存社.克伦特罗小分子半抗原偶联比率的测定方法研究[J].中国饲料, 2005(7):33-35. doi: 10.3969/j.issn.1004-3314.2005.07.013
[19]	林辉, 刘屏, 王建勋, 等.绿原酸人工抗原的合成及反应条件的优化[J].中国中药杂志, 2009, 34(15):1906-1909. doi: 10.3321/j.issn:1001-5302.2009.15.007 LIN H, LIU P, WANG J X, et al. Synthesis and optimize reaction conditions of chlorogenic acid artificial antigen[J]. China Journal of Chinese Materia Medica, 2009, 34(15):1906-1909. doi: 10.3321/j.issn:1001-5302.2009.15.007
[20]	SHAPIRO S M, OUDIZ R J, CAO T, et al. Primary pulmonary hypertension: improved long-term effects and survival with continuous intravenous epoprostenol infusion[J]. Journal of the American College of Cardiology, 1997, 30(2):343-349. doi: 10.1016/S0735-1097(97)00187-3
[21]	吉薇, 郑洁莹, 曾雪萍, 等.南海海域软骨藻酸(DA)贝类毒素的HPLC方法检测[J].现代食品科技, 2011, 27(1):120-122. http://d.old.wanfangdata.com.cn/Periodical/gzspgykj201101030 JI W, ZHENG J Y, ZENG X P, et al. HPLC analysis of domoic acid poisoning in South China Sea[J]. Modern Food Science and Technology, 2011, 27(1):120-122. http://d.old.wanfangdata.com.cn/Periodical/gzspgykj201101030
[22]	吴多加, 李凤琴.软骨藻酸与人类健康关系研究进展[J].卫生研究, 2005, 34(3):378-381. doi: 10.3969/j.issn.1000-8020.2005.03.039 WU J D, LI F Q. Domoic acid and human health[J]. Journal of Hygiene Research, 2005, 34(3):378-381. doi: 10.3969/j.issn.1000-8020.2005.03.039
[23]	SINGH K V, KAUR J, VARSHNEY G C, et al. Synthesis and characterization of hapten-protein conjugates for antibody production against small molecules[J]. Bioconjugate Chemistry, 2004, 15(1):168-173. doi: 10.1021/bc034158v
[24]	SMITH D S, KITTS D D. A competitive enzyme-linked immunoassay for domoic acid determination in human body fluids[J]. Food & Chemical Toxicology, 1994, 32(12):1147-1154. https://www.sciencedirect.com/science/article/pii/0278691594901309
[25]	KAWATSU K, HAMANO Y, NOGUCHI T. Production and characterization of a monoclonal antibody against domoic acid and its application to enzyme immunoassay[J]. Toxicon, 1999, 37(11):1579-1589. doi: 10.1016/S0041-0101(99)00106-3
[26]	YU F Y, CHI T F, LIU B H, et al. Development of a sensitive enzyme-linked immunosorbent assay for the determination of ochratoxin A[J]. Journal of Agricultural & Food Chemistry, 2005, 53(17):6947-6953. http://www.ncbi.nlm.nih.gov/pubmed/16104825
[27]	LITAKER R W, STEWART T N, EBERHART B T L, et al. Rapid enzyme-linked immunosorbent assay for detection of the algal toxin domoic acid[J]. Journal of Shellfish Research, 2008, 27(5):1301-1310. doi: 10.2983/0730-8000-27.5.1301
[28]	王茜, 程金平, 高利利, 等.记忆缺失性贝毒软骨藻酸的污染现状及检测技术[J].安徽农业科学, 2011, 39(26):16070-16073, 16136. doi: 10.3969/j.issn.0517-6611.2011.26.094 WANG Q, CHENG J P, GAO L L, et al. A review for hazards and determination methods of amnesic shellfish poison:domoic acid[J]. Journal of Anhui Agricultural Sciences, 2011, 39(26):16070-16073. doi: 10.3969/j.issn.0517-6611.2011.26.094
[29]	顾佳萍, 袁涛.赤潮毒素软骨藻酸检测方法研究进展[J].海洋通报, 2010, 29(4):472-477. doi: 10.3969/j.issn.1001-6392.2010.04.021 GU J P, YUAN T. A critical review for determination methods of the marine algal toxin-domoic acid[J]. Marine Science Bulletin, 2010, 29(4):472-477. doi: 10.3969/j.issn.1001-6392.2010.04.021
[30]	高利利, 程金平, 刘元嫄, 等.软骨藻酸胶体金免疫层析检测试纸条的研制[J].环境科学, 2011, 32(8):2492-2496. http://d.old.wanfangdata.com.cn/Periodical/hjkx201108054 GAO L L, CHENG J P, LIU Y Y, et al. Development of colloidal gold immunochromatographic strip for rapid detection of domoic acid[J]. Environmental Science, 2011, 32(8):2492-2496. http://d.old.wanfangdata.com.cn/Periodical/hjkx201108054
[31]	陈西平, 王成斌, 胡俊明, 等. HPLC方法检测水及水生动物中软骨藻酸[J].卫生研究, 2001, 30(4):247-248. doi: 10.3969/j.issn.1000-8020.2001.04.022 CHEN X P, WANG C B, HU J M, et al. Determination of domoic acid in water and aquatic animals by high performance liquid chromatography[J]. Journal of Hygiene Research, 2001, 30(4):247-248. doi: 10.3969/j.issn.1000-8020.2001.04.022
[32]	JAMES K J, GILLMAN M, MONICA F A, et al. Amnesic shellfish poisoning toxins in bivalve molluscs in Ireland[J]. Toxicon, 2005, 46(8):852-858. doi: 10.1016/j.toxicon.2005.02.009
[33]	QUILLIAM M A, BROWN N, BATES L A, et al. Determination and confirmation of the amnesic shellfish poisoning toxin, domoic acid, in shellfish from scotland by liquid chromatography and mass spectrometry[J]. Journal of Aoac International, 2001, 84(5):1657-1667. doi: 10.1093/jaoac/84.5.1657
[34]	王恒.舟山海域贝类海产品中软骨藻酸含量调查[J].中国卫生检验杂志, 2011(12):2986-2988. http://www.wanfangdata.com.cn/details/detail.do?_type=perio&id=zgwsjyzz201112067 WANG H. An investigation of domoic acid content in shellfish in Zhoushan islands[J]. Chinese Journal of Health Laboratory Technology, 2011(12):2986-2988. http://www.wanfangdata.com.cn/details/detail.do?_type=perio&id=zgwsjyzz201112067

Cited By

Cited by

Periodical cited type(2)

1.	谢发之，宋恒帅，张道德，杨少华，张梦，方亮，邵永刚. 铅炭电池负极添加剂研究进展. 材料导报. 2023(22): 11-18 .
2.	高鹤，王再红，陈二霞，霍玉龙，孙海涛，闫娜，黄盼盼，张萌，陈志雪. 国产石墨烯应用于铅酸蓄电池的性能研究. 蓄电池. 2021(03): 101-105+132 .

Other cited types(4)

Get Citation

PDF

XML

Article views (1702) PDF downloads (51) Cited by(6)

Turn off MathJax

Article Contents

Abstract

References

Preparation of Polyclonal Antibody Against Domoic Acid and Establishment of Indirect Competitive ELISA Assay

Abstract

References

Cited by

Periodical cited type(2)

Other cited types(4)

Catalog

Export File

Citation

Format

Content