兽药小分子与生物识别元件相互作用研究进展

doi:10.16431/j.cnki.1671-7236.2022.02.038

摘要/Abstract

摘要： 因不合理或违规使用兽药, 导致兽药原型或代谢物在动物源性食品和水体中残留, 继而危害人类健康、破坏生态环境。因此, 兽药残留快速检测方法的建立显得尤为重要。受体、抗体、适配体等生物识别元件具有分子识别能力, 根据其特性已发开了不同的检测方法, 如免疫分析法、受体检测法及生物传感器法等方法, 广泛用于生物医药的研究。由于分子识别元件是分析的主体, 影响分析的选择性, 因此不同识别元件与兽药相互作用的机制越来越受到人们的重视。随着计算机技术的发展, 同源建模和分子对接广泛用于残留检测技术, 主要用于药物与蛋白质之间、抗原与抗体之间、受体与配体之间等生物分子之间相互作用的研究。笔者重点介绍了单链抗体(ScFv)、受体和适配体3种识别元件与兽药小分子之间的互作机制, 主要分析生物识别元件与兽药结合部位形成的氢键、疏水性及关键氨基酸或碱基, 在分子水平上探究药物与识别元件的机理和规律, 从而找出影响识别元件与药物结合的关键因素, 并对3种识别元件进行了总结, 以期为全面了解识别机制和体外进化, 制备出亲和力更广的ScFv、受体、适配体提供理论支撑, 为后续药物残留检测技术和药物开发奠定基础。

关键词: 生物识别元件; 单链抗体(ScFv); 受体; 适配体; 同源建模; 分子对接

Abstract: At present, due to unreasonable or illegal use of veterinary drugs, their prototypes or metabolites remain in animal derived food and water, which will endanger human health and damage the ecological environment.Therefore, the establishment of rapid detection methods of veterinary drug residues is particularly important.Receptors, antibodies, aptamers and other biometric elements have molecular recognition ability.According to its characteristics, different detection methods have been developed such as immunoassay, receptor detection and biosensor method, which are widely used in biomedical research.Because molecular recognition elements are the main body of analysis and affect the selectivity of analysis, the mechanism of interaction between different recognition elements and veterinary drugs has been paid more and more attention.With the development of computer technology, homologous modeling and molecular docking are widely used in residue detection, mainly used in the study of the interactions between drugs and proteins, antigens and antibodies, receptors and ligands and other biomolecules.This paper focused on the interaction mechanism between three recognition elements of single chain variable fragment (ScFv), receptor and aptamer and veterinary drug small molecules.It mainly analyzed the hydrogen bonds, hydrophobicity and key amino acids or bases formed at the binding site of biometric elements and veterinary drugs.The mechanism and law of drug and recognition element were explored at molecular level, so as to find out the key factors affecting the combination of drug and recognition element, and three recognition elements were summarized, in order to provide theoretical support for the comprehensive understanding of recognition mechanism and in vitro evolution, and prepare ScFv, receptors and aptamers with wider affinity, and lay a foundation for subsequent drug residue detection technology and drug development.

Key words: biometric element; single chain variable fragment (ScFv); receptor; aptamer; homology modeling; molecular docking

中图分类号:

S859

吴双敏, 李龙, 侯仁, 陈玉霜, 彭大鹏. 兽药小分子与生物识别元件相互作用研究进展[J]. 中国畜牧兽医, 2022, 49(2): 755-764.

WU Shuangmin, LI Long, HOU Ren, CHEN Yushuang, PENG Dapeng. Research Progress on the Interaction Between Small Molecules of Veterinary Drugs and Biometric Elements[J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(2): 755-764.

参考文献

[1] 郭思依, 孙明娜, 董旭, 等. 免疫分析技术在农药残留快速检测中的应用及研究进展[J]. 中国农学通报, 2021, 37(23): 106-112.
GUO S Y, SUN M N, DONG X, et al. Application and research progress of immunoassay technology in rapid detection of pesticide residues[J]. Chinese Agricultural Science Bulletin, 2021, 37(23): 106-112. (in Chinese)
[2] 何扩, 张秀媛, 杜欣军, 等. 基因工程抗体在食品安全检测中应用进展研究[J]. 中国粮油学报, 2014, 29(8): 124-128.
HE K, ZHANG X Y, DU X J, et al. Application progress of genetic engineering antibody in food safety testing[J]. Journal of the Chinese Cereals and Oils Association, 2014, 29(8): 124-128. (in Chinese)
[3] 于昊天. 全人源抗蓖麻毒素单链抗体制备及其生物学活性研究[D]. 北京: 军事科学院, 2020.
YU H T. Preparation of full-human anti-RT single-chain antibody and study on its biological activity[D]. Beijing: Academy of Military Sciences, 2020. (in Chinese)
[4] 张泽, 张颖聪, 于洪伟, 等. 生物传感器识别元件的种类及其在临床检验中的研究进展[J]. 临床检验杂志, 2020, 38(10): 767-771.
ZHANG Z, ZHANG Y C, YU H W, et al. Biosensor identification elements and their research progress in clinical laboratory[J]. Journal of Clinical Laboratory, 2020, 38(10): 767-771. (in Chinese)
[5] 李群林. 分子模拟技术研究吡唑乙基苯甲酰胺衍生物作为食欲素受体1拮抗剂[D]. 上海: 上海应用技术大学, 2020.
LI Q L. Molecular modeling technology studies of novel pyrazoylethylbenzamide derivatives as selective orexin oeceptor 1 antagonists[D]. Shanghai: Shanghai Institute of Technology, 2020. (in Chinese)
[6] 林婷, 黄义德. 单链抗体在毕赤酵母中表达的研究进展[J]. 生物技术通讯, 2019, 30(5): 693-701.
LIN T, HUANG Y D. Research progress of the expression of single-chain variable fragment in Pichia pastoris[J]. Letters in Biotechnology, 2019, 30(5): 693-701. (in Chinese)
[7] AGUIAR R B, SILVA T A, COSTA B A, et al. Generation and functional characterization of a single-chain variable fragment (ScFv) of the anti-FGF2 3F12E7 monoclonal antibody[J]. Science Reports, 2021, 11(1): 1432.
[8] ChRISTOPH E H, CONSTANT Z M, DOMINIK E, et al. Single-chain antibodies as diagnostic tools and therapeutic agents[J]. Intravascular Biology Meeting, 2008, 101(6): 1012-1019.
[9] HE X, DUAN C F, QI Y H, et al. Virtual mutation and directional evolution of anti-amoxicillin ScFv antibody for immunoassay of penicillins in milk[J]. Analytical and Bioanalytical Chemistry, 2017, 517: 9-17.
[10] XIE S, WEN K, XIE J, et al. Magnetic-assisted biotinylated single-chain variable fragment antibody-based immunoassay for amantadine detection in chicken[J]. Analytical and Bioanalytical Chemistry, 2018, 410(24): 6197-6205.
[11] ZHANG X, HE K, ZHAO R, et al. Cloning of ScFv from hybridomas using a rational strategy: Application as a receptor to sensitive detection microcystin-LR in water[J]. Chemosphere, 2016, 160: 230-236.
[12] ZHAO F C, TIAN Y, WANG H M, et al. Development of a biotinylated broad-specificity single-chain variable fragment antibody and a sensitive immunoassay for detection of organophosphorus pesticides[J]. Analytical and Bioanalytical Chemistry, 2016, 408(23): 6423-6430.
[13] 尹志安. 猪流行性腹泻病毒野生株CH/JX226/2018的分离培养及抗病毒单链抗体研究[D]. 郑州: 河南农业大学, 2019.
YIN Z A. The culture isolation of wild-type PEDV CH/JX226/2018 and the research on anti-PEDV single-chain antibody[D]. Zhengzhou: Henan Agricultural University, 2019. (in Chinese)
[14] AHMED S, NING J, CHENG G, et al. Receptor-based screening assays for the detection of antibiotics residues——A review[J]. Talanta, 2017, 166: 176-186.
[15] 殷萌琪. α-玉米赤霉醇类化合物和β-内酰胺类抗生素的广谱快速检测方法研究[D]. 无锡: 江南大学, 2020.
YIN M Q. Study on broad-spectrum and rapid determination of α-zearalanol compounds and β-lactam antibiotics[D]. Wuxi: Jiangnan University, 2020. (in Chinese)
[16] MOON J, KIM G, PARK S B, et al. Comparison of whole-cell SELEX methods for the identification of Staphylococcus aureus-specific DNA aptamers[J]. Sensors, 2015, 15(4): 8884-8897.
[17] 崔妍, 白亚龙, 史贤明. 核酸适配体在金黄色葡萄球菌检测中的应用进展[J]. 食品工业科技, 2021, 42(21): 1-7.
CUI Y, BAI Y L, SHI X M. Progress on the application of aptamers in the detection of Staphylococcus aureus[J]. Science and Technology of Food Industry, 2021, 42(21): 1-7. (in Chinese)
[18] WANG J L, LU T T, HU Y, et al. A label-free and carbon dots based fluorescent aptasensor for the detection of kanamycin in milk[J]. Spectrochimica Acta. Part A, Molecular and Biomolecular Spectroscopy, 2020, 226: 117651.
[19] ZON G. Mini-review: Recent advances in aptamer applications[J]. Journal of Cancer Treatment and Diagnosis, 2020, 4(3): 113894
[20] LI F Q, YU Z G, HAN X D, et al. Electrochemical aptamer-based sensors for food and water analysis: A review[J]. Analytica Chimica Acta, 2019, 1051: 1-23.
[21] TANG Z Y, JUN Y G, LV Y J, et al. Aptamer-conjugated and doxorubicin-loaded grapefruit-derived nanovectors for targeted therapy against HER2⁺ breast cancer[J]. Journal of Drug Targeting, 2020, 28(2): 186-194.
[22] 刘蕊, 农药特异性基因工程抗体的分子设计、表达及识别机制[D]. 杭州: 浙江大学, 2016.
LIU R. Molecular design expression and recongnition mechanism of engingeering antibodies against pesticides[D]. Hangzhou: Zhejiang University, 2016. (in Chinese)
[23] 陈烨. 基于群智能算法的蛋白质结构预测研究[D]. 北京: 中国矿业大学, 2018.
CHEN Y. Study on the protein structure prediction based on swarm intelligence algorithm[D]. Beijing: China University of Mining and Technology, 2018. (in Chinese)
[24] 仝泽方. 通过氨基酸定点突变调控水稻OsHAL3蛋白功能的研究[D]. 沈阳: 沈阳农业大学, 2018.
TONG Z F. Regulation of the OsHAL3 protein function by site-directed mutagenesis of amino acids[D]. Shenyang: Shenyang Agricultural University, 2018. (in Chinese)
[25] 王鸽. 四环素类药物受体的制备及进化[D]. 保定: 河北农业大学, 2020.
WANG G. Preparation and directional evolution of the receptor of tetracyclines[D]. Baoding: Hebei Agricultural University, 2020. (in Chinese)
[26] 成胜荣. 同源异效桑源药材(桑叶、桑枝、桑白皮、桑椹)的物质基础研究[D]. 镇江: 江苏大学, 2019.
CHENG S R. Study on the material basis of homologous and heterologous mulberry medicinal materials (mulberry leaves, mulberry branches, mulberry white skin and mulberry)[D]. Zhenjiang: Jiangsu University, 2019. (in Chinese)
[27] LIU Y H, JIN M J, GUI J W, et al. Hapten design and indirect competitive immunoassay for parathion determination: Correlation with molecular modeling and principal component analysis[J]. Analytical and Bioanalytical Chemistry, 2007, 591(2): 173-182.
[28] LIU M, ZHENG N, LI D, et al. Cyp51A-based mechanism of azole resistance in Aspergillus fumigatus: Illustration by a new 3D structural model of Aspergillus fumigatus CYP51A protein[J]. Medical Mycology, 2016, 54(4): 400-408.
[29] 宋亚宁, 胡超琼, 王冲, 等. 核酸适配体生物传感器在食品中氟喹诺酮类兽药残留检测中的应用[J]. 中国食品学报, 2021, 21(8): 409-419.
SONG Y N, HU C Q, WANG C, et al. Application of aptamer biosensor in the determination of fluoroquinolones residues in food[J]. Journal of Chinese Institute of Food Science and Technology, 2021, 21(8): 409-419. (in Chinese)
[30] WANG J P, DONG J, DUAN C F, et al. Production and directional evolution of antisarafloxacin ScFv antibody for immunoassay of fluoroquinolones in milk[J]. Agricultural Food Chemistry, 2016, 64(42): 7957-7965.
[31] 曹子健, 何欣, 王建平, 等. 抗沙拉沙星ScFv同源模拟及分子结合机制[J]. 黑龙江畜牧兽医, 2018, 1: 47-51.
CAO Z J, HE X, WANG J P, et al. Homology modeling of anti-salafloxacin ScFv antibody and the mechanism of molecular binding[J]. Heilongjiang Animal Science and Veterinary Medicine, 2018, 1: 47-51. (in Chinese)
[32] ZHANG X Y, HE K, ZHANG D H, et al. Production and characterization of a monoclonal antibody for pefloxacin and mechanism study of antibody recognition[J]. Bioscience, Biotechnology, and Biochemistry, 2019, 83(4): 633-640.
[33] WEN K, NOLKE G, SCHILLBERG S, et al. Improved fluoroquinolone detection in ELISA through engineering of a broad-specific single-chain variable fragment binding simultaneously to 20 fluoroquinolones[J]. Analytical and Bioanalytical Chemistry, 2012, 403(9): 2771-2783.
[34] 董文婷, 曾勇, 吴晓翠, 等. 液相色谱-串联质谱法测定禽蛋中β-内酰胺类药物残留[J]. 食品安全质量检测学报, 2020, 11(20): 7582-7590.
DONG W T, ZENG Y, WU X C, et al. Determination of β-lactam in poultry eggs by liquid chromatography tandem mass spectrometry[J]. Journal of Food Safety and Quality, 2020, 11(20): 7582-7590. (in Chinese)
[35] 王俊豪, 丛萌, 陶燕飞, 等. 鸡蛋中抗菌药物残留消除规律研究进展[J]. 中国抗生素杂志, 2020, 45(12): 1208-1220.
WANG J H, CONG M, TAO Y F, et al. Research progress on elimination of antimicrobial residues in eggs[J]. Chinese Journal of Antibiotics, 2020, 45(12): 1208-1220. (in Chinese)
[36] LIU J, ZHANG H C, DUAN C F, et al. Production of anti-amoxicillin ScFv antibody and simulation studying its molecular recognition mechanism for penicillins[J]. Environmental Science and Health B, 2016, 51(11): 742-750.
[37] 段长飞. 阿莫西林单链抗体的定向进化[D]. 保定: 河北农业大学, 2017.
DUAN C F. Directional evolution of the anti-amoxicillin single chain antibody fragment[D]. Baoding: Hebei Agricultural University, 2017. (in Chinese)
[38] XIE S L, WANG J Y, YU X Z, et al. Site-directed mutations of anti-amantadine ScFv antibody by molecular dynamics simulation: Prediction and validation[J]. Molecular Modeling, 2020, 26(3): 49.
[39] LI L, HOU R, WANG X Q, et al. Development of a monoclonal-based ic-ELISA for the determination of kitasamycin in animal tissues andsimulation studying its molecular recognition mechanism[J]. Food Chemistry, 2021, 363: 129465.
[40] 史芳舒. 吩噻嗪类药物单链抗体的制备及进化[D]. 保定: 河北农业大学, 2018.
SHI F S. Preparation and directional evolution of the anti-phenothiazines single chain antibody fragment[D]. Baoding: Hebei Agricultural University, 2018. (in Chinese)
[41] LU Q, HOU Y Y, LIU X X, et al. Construction, expression and functional analysis of anti-clenbuterol codon-optimized ScFv recombinant antibody[J]. Food Chemical Toxicology, 2020, 135: 110973.
[42] LIANG X, LI C, ZHU J, et al. Dihydropteroate synthase based sensor for screening multi-sulfonamides residue and its comparison with broad-specific antibody based immunoassay by molecular modeling analysis[J]. Analytica Chimica Acta 2019, 1050: 139-145.
[43] NING J N, AHMED S, CENG G Y, et al. Analysis of the stability and affinity of BlaR-CTD protein to β-lactam antibiotics based on docking and mutagenesis studies[J]. Biological Engineering, 2019, 13: 27.
[44] WANG G, ZHANG H C, LIU J, et al. A receptor-based chemiluminescence enzyme linked immunosorbent assay for determination of tetracyclines in milk[J]. Analytical Biochemistry, 2019, 564-565: 40-46.
[45] WANG G, XIA W Q, LIU J X, et al. Directional evolution of TetR protein and development of a fluoroimmunoassay for screening of tetracyclines in egg[J]. Microchemical Journal, 2019, 150: 104184.
[46] ZHANG J, ZHANG T H, GUAN T Z, et al. Spectroscopic and molecular modeling approaches to investigate the interaction of bisphenol A, bisphenol F and their diglycidyl ethers with PPARα[J]. Chemosphere, 2017, 180: 253-258.
[47] GUAN T Z, SUN Y H, WANG Y J, et al. Multi-residue method for the analysis of stilbene estrogens in milk[J]. Molecular Sciences, 2019, 20(3): 44.
[48] RAZA M, AHMAD A, YUE F, et al. Biophysical and molecular docking approaches for the investigation of biomolecular interactions between amphotericin B and bovine serum albumin[J]. Photochemistry and Photobiology. B, Biology, 2017, 170: 6-15.
[49] ZHAO T, LIU Z H, NIU J M, et al. Investigation of the interaction mechanism between salbutamol and human serum albumin by multispectroscopic and molecular docking[J]. BioMed Research International, 2020, 2020: 1693602.
[50] MENG X Y, ZHANG H X, MEZEI M, et al. Molecular docking: A powerful approach for structure-based drug discovery[J]. Current Computer-Aided Drug Design, 2011, 7(2): 146-157.
[51] SONG M C, LIM S J, TONG J C. Recent advances in computer-aided drug design[J]. Briefings in Bioinformatics, 2009, 10(5): 579-591.
[52] YANG J T, LI Q, BIAN L J. Spectroscopic analysis and docking simulation on the recognition and binding of TEM-1 β-lactamase with β-lactam antibiotics[J]. Experimental and Therapeutic Medicine, 2017, 14: 3288-3298.
[53] LI Q, ZHANG T L, BIAN L J. Recognition and binding of β-lactam antibiotics to bovine serum albumin by frontal affinity chromatography in combination with spectroscopy and molecular docking[J]. Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences, 2016, 1014: 90-101.
[54] 梁晓. 基于二氢叶酸合成酶的磺胺类药物多残留快速检测分析研究[D]. 北京: 中国农业大学, 2014.
LIANG X. Development of rapid detection methods based on dihydropteroate synthase for sulfonamides[D]. Beijing: China Agricultural University, 2014. (in Chinese)
[55] 关天竺. 基于雌激素受体的双酚类化合物检测方法及构效关系研究[D]. 长春: 吉林大学, 2019.
GUAN T Z. Estrogen receptor-based multi-residue screening of bisphenol compounds and structure-activity relationships studies[D]. Changchun: Jilin University, 2019. (in Chinese)
[56] ZHANG Q, NI Y. Comparative studies on the interaction of nitrofuran antibiotics with bovine serum albumin[J]. Royal Society of Chemistry, 2017, 7(63): 39833-39841.
[57] PENG W, DING F, PENG Y K, et al. Molecular recognition of malachite green by hemoglobin and their specific interactions: Insights from in silico docking and molecular spectroscopy[J]. Molecular BioSystems, 2014, 10(1): 138-148.
[58] LI Z, BO P, ZHANG H M. Investigation on the interaction between an antimicrobial in aquaculture, malachite green and hemocyanin from mud crab Scylla paramamosain[J]. Molecular and Biomolecular Spectroscopy, 2015, 135: 669-675.
[59] 韩旭艳. 氨基糖苷类抗生素特异性单链DNA适配体的筛选及其应用研究[D]. 无锡: 江南大学, 2018.
HAN X Y. Selection and application of aminoglycoside antibiotics-specific single-stranded DNA aptamers[D]. Wuxi: Jiangnan University, 2018. (in Chinese)
[60] SADEGHI A S, MOHSENZADEH M, ABNOUS K, et al. Development and characterization of DNA aptamers against florfenicol: Fabrication of a sensitive fluorescent aptasensor for specific detection of florfenicol in milk[J]. Talanta, 2018, 182: 193-201.
[61] YANG L, NI H J, LI C L, et al. Development of a highly specific chemiluminescence aptasensor for sulfamethazine detection in milk based on in vitro selected aptamers[J]. Sensors and Actuators B: Chemical, 2019, 281: 801-811.
[62] 邹雪梅. 阿米卡星适配体的筛选及在残留检测中的应用[D]. 镇江: 江苏大学, 2018.
ZOU X M. Selection of amikacin aptamer and its application in the detection of amikacin residue[D]. Zhenjiang: Jiangsu University, 2018. (in Chinese)
[63] 张玉红. 妥布霉素特异性单链DNA适配体的筛选及其序列优化和应用研究[D]. 无锡: 江南大学, 2017.
ZHANG Y H. Selection and sequence optimization of tobamycin specific single stranded DNA aptamers and their applications[D]. Wuxi: Jiangnan University, 2017. (in Chinese)
[64] EISOLD A, LABUDDE D. Detailed analysis of 17β-estradiol-aptamer interactions: A molecular dynamics simulation study[J]. Molecules, 2018, 23(7): 1690.