Preparation and Application of Monoclonal Antibody against Etomidate

doi:10.13523/j.cb.2307037

China Biotechnology

2023, Vol. 43

Issue (11): 27-34 DOI: 10.13523/j.cb.2307037

Preparation and Application of Monoclonal Antibody against Etomidate

CHEN Fei^1,²,WU Li-xian^2,^**(

),GUO Jin-chen²,LIANG Fei-min²

1 Guangzhou Accurate and Correct Test Company, Guangzhou 510663, China
2 Guangzhou Wondfo Biotech Co., Ltd,Guangzhou 510663,China

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Abstract Objective: To prepare anti-ET antibody for rapid detection of ET using the colloidal gold method. Methods: After identified by mass spectrometry and ultraviolet spectrum, the synthetic ET antigen was used to immunize Balb/C mice. ET hybridoma cell lines with high specificity were obtained by hybridoma technique. After purification by Protein G column, the antibody with appropriate sensitivity was selected and applied to ET colloidal gold reagent. The sensitivity and specificity of the antibody were identified by drug cross interference experiment and clinical experiment. Results: The antibody produced by 10F9 cell line was suitable for ET colloidal gold immunochromatography reagent. The ET reagent was used to detect 12 kinds of e-liquid additives and 48 kinds of commonly used drugs without cross-reaction. The cutoff of ET reagent in urine and e-liquid were both 500 ng/mL, the sensitivity was 100%, and the specificity was 99.5%.Conclusions: The monoclonal antibody (mAb) against ET was successfully prepared and applied to the colloidal gold immunochromatographic reagent.

Key words： Etomidate Monoclonal antibodies Colloidal gold immunochromatography

Received: 31 July 2023 Published: 01 December 2023

ZTFLH:

Q51

Corresponding Authors: **Li-xian WU E-mail: lillian7040@126.com

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	CHEN Fei
	WU Li-xian
	GUO Jin-chen
	LIANG Fei-min

Cite this article:

CHEN Fei, WU Li-xian, GUO Jin-chen, LIANG Fei-min. Preparation and Application of Monoclonal Antibody against Etomidate. China Biotechnology, 2023, 43(11): 27-34.

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https://manu60.magtech.com.cn/biotech/10.13523/j.cb.2307037 OR https://manu60.magtech.com.cn/biotech/Y2023/V43/I11/27

Fig.1 Structure diagram of ET hapten

Fig.2 Diagram of glutaraldehyde coupling R: ET hapten;P: Carrier protein

Fig.3 Schematic diagram of colloidal gold reagent strips

Fig.4 Ethyl 1-(1-(4-aminophenyl)ethyl)-1H-imidazole-5-carboxylate ESI-MS

Fig.5 UV spectra of ET complete antigen and their carrier protein A. UV spectra of ET-KLH and KLH protein B. UV spectra of ET-BSA and BSA protein

Fig.6 Titer and inhibition rate of ET mAbs A.Titer of the ET mAbs B. Inhibition of the ET mAbs

Fig.7 SDS-PAGE for the mAb of 10F9 cell line Line 1:Marker;Line 2:10F9 mAbs

Fig.8 Cutoff values of ET reagent strips A. Cutoff values of ET reagent strips in urine B. Cutoff values of ET reagent strips in e-liquid “+”means positive,“-”means negative

Table 1 Results of cross experiments on 12 kinds of commonly used additives in e-liquid and 48 kinds of commonly used drugs

Table 2 Test results of clinical sample

Table 3 Clinical test results of ET reagent on e-liquid


[1]	徐若沦, 解通, 胡庆坤, 等. 依托咪酯药理作用、药物代谢动力学及毒理学研究. 广东公安科技, 2022, 30(2):43-45.

[1]	Xu R L, Xie T, Hu Q K, et al. Pharmacological action, pharmacokinetics and toxicology of etomidate. Guangdong Gongan Keji, 2022, 30(2):43-45.

[2]	王辉, 岳彬, 郝晓燕. 依托咪酯应用于心血管患者麻醉诱导的临床疗效观察. 当代医学, 2021, 27(3): 62-64.

[2]	Wang H, Yue B, Hao X Y. Observation of the clinical efficacy of etomidate applied to anesthesia induction in cardiovascular patients. Contemporary Medicine, 2021, 27(3): 62-64.

[3]	李悦. 老年人性别差异与全身麻醉药用药剂量的相关研究. 长春: 吉林大学, 2021.

[3]	Li Y. Correlation between gender difference and dosage of general anesthetics in the elderly. Changchun: Jilin University, 2021.

[4]	中华人民共和国公安部. 2021年中国毒情形势报告.[2023-07-07].https://www.mps.gov.cn/n2255079/n6865805/n7355741/n7355780/c8660792/content.html.

[4]	Ministry of Public Security of the China.China drug situation report 2021.[2023-07-07].https://www.mps.gov.cn/n2255079/n6865805/n7355741/n7355780/c8660792/content.html.

[5]	王太云, 刘宜平, 李恩有. 依托咪酯常见不良反应及预防新策略研究进展. 中国医药, 2021, 16(9): 1411-1414.

[5]	Wang T Y, Liu Y P, Li E Y. Research progress on common adverse reactions of etomidate and new prevention strategies. China Medicine, 2021, 16(9): 1411-1414.

[6]	Valk B I, Struys M M R F. Etomidate and its analogs: a review of pharmacokinetics and pharmacodynamics. Clinical Pharmacokinetics, 2021, 60(10): 1253-1269. doi: 10.1007/s40262-021-01038-6 pmid: 34060021

[7]	Jung Y K, You S Y, Kim S Y, et al. Simultaneous determination of etomidate and its major metabolite, etomidate acid, in urine using dilute and shoot liquid chromatography-tandem mass spectrometry. Molecules (Basel, Switzerland), 2019, 24(24): 4459. doi: 10.3390/molecules24244459

[8]	Yum H, Jeong S, Jang M, et al. Fast and reliable analysis of veterinary metomidate and etomidate in human blood samples by liquid chromatography-tandem mass spectrometry (LC-MS/MS) in a postmortem case. Journal of Forensic Sciences, 2021, 66(6): 2532-2538. doi: 10.1111/jfo.v66.6

[9]	Park Y J, Cho E, Kim S H, et al. Determination of etomidate and etomidate acid in hair using liquid chromatography-tandem mass spectrometry. Journal of Forensic Sciences, 2022, 67(6): 2479-2486. doi: 10.1111/jfo.v67.6

[10]	许俊丽, 刘贝贝, 王玉龙, 等. 胶体金免疫层析试纸法检测农产品中戊唑醇残留. 分析化学, 2019, 47(11):1823-1831.

[10]	Xu J L, Liu B B, Wang Y L, et al. Lateral flow assay for determination of tebuconazole in agricultural products. Chinese Journal of Analytical Chemistry, 2019, 47(11):1823-1831.

[11]	康可人, 刘锋, 谭仙桂, 等. 抗美沙酮单克隆抗体制备、鉴定和在侧向免疫层析方法中的应用. 中国药物滥用防治杂志, 2013, 19(1): 14-19.

[11]	Kang K R, Liu F, Tan X G, et al. Development of monoclonal antibodies against methadone and its application in lateral flow immunoassay. Chinese Journal of Drug Abuse Prevention and Treatment, 2013, 19(1): 14-19.

[12]	卢乐华, 茹洁雄, 李祥林, 等. 电子烟液中游离态和质子态烟碱的测定及分布. 烟草科技, 2021, 54(8):36-44.

[12]	Lu L H, Ru J X, Li X L, et al. Determination and distribution of free-base and protonated nicotine in e-liquids. Tobacco Science & Technology, 2021, 54(8):36-44.

[13]	杨文武, 况利平. 电子烟烟液中甜味剂的种类及感官评价现状探究. 食品工业, 2023, 44(4): 200-204.

[13]	Yang W W, Kuang L P. Study on the types and sensory evaluation of sweeteners in E-cigarette liquid. The Food Industry, 2023, 44(4): 200-204.

[14]	郭乃菲, 于基成, 马福敏, 等. 常用人工抗原鉴定方法的比较. 食品与生物技术学报, 2014, 33(5):517-521.

[14]	Guo N F, Yu J C, Ma F M, et al. Study of comparison the commonly used artificial antigen identification methods. Journal of Food Science and Biotechnology, 2014, 33(5):517-521.

[15]	宋娟, 王榕妹, 王悦秋, 等. 半抗原的设计、修饰及人工抗原的制备. 分析化学, 2010, 38(8):1211-1218. doi: 10.1016/S1872-2040(09)60063-3

[15]	Song J, Wang R M, Wang Y Q, et al. Hapten design, modification and preparation of artificial antigens. Chinese Journal of Analytical Chemistry, 2010, 38(8):1211-1218. doi: 10.1016/S1872-2040(09)60063-3

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