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The Natural Products Journal

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ISSN (Print): 2210-3155
ISSN (Online): 2210-3163

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Research Article

Sandwich Enzyme-linked Immunosorbent Assay (ELISA) to Quantify Monoclonal Antibody (B[a]P-13) for Herbal Medicine Products

Author(s): Yong-Yeon Kim and Han-Seung Shin*

Volume 13, Issue 3, 2023

Published on: 13 December, 2022

Article ID: e041122210658 Pages: 9

DOI: 10.2174/2210315513666221104154116

Price: $65

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Abstract

Sandwich enzyme-linked immunosorbent assay (ELISA) to quantify monoclonal antibody (B[a]P-13).

Background: Only a few studies have focused on the analysis using specific antibodies in the sandwich ELISA method to each B[a]P in herbal medicine products. In contrast to the sandwich ELISA method, many competitive ELISA methods using specific antibodies such as benzo[a]pyrene monoclonal antibody (B[a]P-13) and a goat anti-mouse IgG (H+L) cross-adsorbed secondary antibody, horseradish peroxidase (HRP) were developed.

Introduction: The objective of this study was to develop and validate the method for the response of the benzo[a]pyrene monoclonal antibody (B[a]P-13) and goat anti-mouse IgG (H+L) cross-adsorbed secondary antibody (HRP) to prepare the immunogen and its application to detect the benzo[a]pyrene in various herbal medicine products.

Methods: This research method includes preparation of B[a]P-protein conjugates, sampling and extraction procedure for herbal medicines, sandwich ELISA procedure, evaluation of cross-reactivity for determination, matrix effect of the organic solvents, correlation of benzo[a]pyrene detection ELISA compared to HPLC-FLD in herbal medicine products.

Results: The sandwich ELISA method for B[a]P was validated in linearity (R2 > 0.99), the limit of detection (LOD) (0.08-0.19 μg/kg) and limit of quantification (LOQ) (0.24-0.57 μg/kg), accuracy (95.58-117.06%), and precision (3.80-10.26%). The cross-reactivity (CR) was found for B[a]P (100%), CHR (39%), B[b]F (27%), and B[a]A (41%). As a solvent, acetonitrile (MeCN) was used to express the normalized sandwich ELISA calibration curves with benzo[a]pyrene monoclonal antibody (B[a]P-13). The antigen-antibody binding in sandwich ELISA was decreased about 10 times with increasing the salt content (0.006-0.18 mol/L phosphate to 20-400 mmol/L). The pH range from 6 to 9 was not considered to affect the performance of the sandwich ELISA. Correlation of B[a]P detection in herbal medicines with ELISA compared to HPLC-FLD expressed good correlation (R2 = 0.991) and the slope of the graph for the ELISA (B[a]P-equivalents μg/kg) value divided by the HPLC-FLD (B[a]P μg/kg) value was 0.7292.

Conclusion: Therefore, sandwich ELISA method using benzo[a]pyrene monoclonal antibody (B[a]P- 13) could be an alternative screening method for detection of B[a]P in herbal medicine products.

Graphical Abstract

[1]
Grimmer, G.; Jacob, J.; Dettbarn, G.; Naujack, K.W.; Heinrich, U. Urinary metabolite profile of PAH as a potential mirror of the genetic disposition for cancer. Exp. Toxicol. Pathol., 1995, 47(6), 421-427.
[http://dx.doi.org/10.1016/S0940-2993(11)80319-2] [PMID: 8871077]
[2]
Grimmer, G.; Jacob, J.; Dettbarn, G.; Naujack, K.W. Determination of urinary metabolites of polycyclic aromatic hydrocarbons (PAH) for the risk assessment of PAH-exposed workers. Int. Arch. Occup. Environ. Health, 1997, 69(4), 231-239.
[http://dx.doi.org/10.1007/s004200050141] [PMID: 9137996]
[3]
Ministry of Food and Drug Safety (MFDS). Korean pharmacopoeia and other herbal medicine (crude medicine) specifications. , 2007. Available from: https://www.mfds.go.kr/eng/brd/m_18/view.do?seq=70483&srchFr=&srchTo=&srchWord=&srchTp=&itm_seq_1=0&itm_seq_2=0&multi_itm_seq=0&company_cd=&company_nm=&page=1
[4]
Nikolaou, K.; Masclet, P.; Mouvier, G. PAH stability scale established in situ in an urban region. Sci. Total Environ., 1984, 36, 383-388.
[http://dx.doi.org/10.1016/0048-9697(84)90291-2]
[5]
Farmer, P.B.; Singh, R.; Kaur, B.; Sram, R.J.; Binkova, B.; Kalina, I.; Popov, T.A.; Garte, S.; Taioli, E.; Gabelova, A.; Cebulska-Wasilewska, A. Molecular epidemiology studies of carcinogenic environmental pollutants. Mutat. Res. Rev. Mutat. Res., 2003, 544(2-3), 397-402.
[http://dx.doi.org/10.1016/j.mrrev.2003.09.002] [PMID: 14644342]
[6]
Ministry of Food and Drug Safety (MFDS). Benzo[a]pyrene standards and test methods for sukjihwang and dried jihwang among herbal medicines. Partial revision of food standards and specifications, 2009. Available from: https://www.mfds.go.kr/eng/brd/m_18/view.do?seq=70483&srchFr=&srchTo=&srchWord=&srchTp=&itm_seq_1=0&itm_seq_2=0&multi_itm_seq=0&company_cd=&company_nm=&page=1
[7]
Conney, A.H.; Miller, E.C.; Miller, J.A. Substrate-induced synthesis and other properties of benzpyrene hydroxylase in rat liver. J. Biol. Chem., 1957, 228(2), 753-766.
[http://dx.doi.org/10.1016/S0021-9258(18)70657-1] [PMID: 13475357]
[8]
Hwang, H.J.; Lee, S.H.; Kim, Y.Y.; Shin, H.S. Polycyclic aromatic hydrocarbon risk assessment and analytical methods using QuEchERS pretreatment for the evaluation of herbal medicine ingredients in Korea. Foods, 2021, 10(9), 2200-2214.
[http://dx.doi.org/10.3390/foods10092200] [PMID: 34574309]
[9]
Knopp, D.; Seifert, M.; Väänänen, V.; Niessner, R. Determination of polycyclic aromatic hydrocarbons in contaminated water and soil samples by immunological and chromatographic methods. Environ. Sci. Technol., 2000, 34(10), 2035-2041.
[http://dx.doi.org/10.1021/es991215f]
[10]
Ding, Y.S.; Chou, T.; Abdul-Salaam, S.; Hearn, B.; Watson, C.H. Development of a method to estimate mouth-level benzo[a]pyrene intake by filter analysis. Cancer Epidemiol. Biomarkers Prev., 2012, 21(1), 39-44.
[http://dx.doi.org/10.1158/1055-9965.EPI-11-0800] [PMID: 22028404]
[11]
Corps Ricardo, A.I.; Guzmán Bernardo, F.J.; Zougagh, M.; Rodríguez Martín-Doimeadios, R.C.; Ríos, Á. Magnetic nanoparticles-Carbon nanotubes hybrid composites for selective solid-phase extraction of polycyclic aromatic hydrocarbons and determination by ultra-high performance liquid chromatography. Anal. Bioanal. Chem., 2017, 409(21), 5125-5132.
[http://dx.doi.org/10.1007/s00216-017-0459-7] [PMID: 28714042]
[12]
Zielinski, T.L.; Smith, S.A.; Pestka, J.J.; Gray, J.I.; Smith, D.M. Elisa to quantify hexanal-protein adducts in a meat model system. J. Agric. Food Chem., 2001, 49(6), 3017-3023.
[http://dx.doi.org/10.1021/jf001151o] [PMID: 11410003]
[13]
Goodridge, C.F.; Beaudry, R.M.; Pestka, J.J.; Smith, D.M. ELISA for monitoring lipid oxidation in chicken myofibrils through quantification of hexanal-protein adducts. J. Agric. Food Chem., 2003, 51(26), 7533-7539.
[http://dx.doi.org/10.1021/jf034553f] [PMID: 14664503]
[14]
Fan, Z.; Li, Z.; Liu, S.; Yang, F.; Bian, Z.; Wang, Y.; Tang, G.; Zhao, Q.; Deng, H.; Liu, S. Rapid fluorescence immunoassay of benzo[a]pyrene in mainstream cigarette smoke based on a dual-functional antibody–DNA conjugate. RSC Advances, 2018, 8(52), 29562-29569.
[http://dx.doi.org/10.1039/C8RA04915G] [PMID: 35547323]
[15]
Matschulat, D.; DengOn, A.; Niessner, R.; Knopp, D. Development of a highly sensitive monoclonal antibody based ELISA for detection of benzo[a]pyrene in potable water. Analyst, 2005, 130(7), 1078-1086.
[http://dx.doi.org/10.1039/b503636d] [PMID: 15965533]
[16]
Scharnweber, T.; Fisher, M.; Suchànek, M.; Knopp, D.; Niessner, R. Monoclonal antibody to polycyclic aromatic hydrocarbons based on a new benzo[a]pyrene immunogen. Fresenius J. Anal. Chem., 2001, 371(5), 578-585.
[http://dx.doi.org/10.1007/s002160101012] [PMID: 11767882]
[17]
Fritcher, D.L.; Mazet, J.A.K.; Ziccardi, M.H.; Gardner, I.A. Evaluation of two direct immunoassays for rapid detection of petroleum products on marine birds. Mar. Pollut. Bull., 2002, 44(5), 388-395.
[http://dx.doi.org/10.1016/S0025-326X(01)00247-8] [PMID: 12146821]
[18]
Schedl, M.; Wilharm, G.; Achatz, S.; Kettrup, A.; Niessner, R.; Knopp, D. Monitoring polycyclic aromatic hydrocarbon metabolites in human urine: Extraction and purification with a sol-gel glass immunosorbent. Anal. Chem., 2001, 73(23), 5669-5676.
[http://dx.doi.org/10.1021/ac010868n] [PMID: 11774906]
[19]
Santella, R.M.; Lin, C.D.; Dharmaraja, N. Monoclonal antibodies to a benzo[a]pyrene diolepoxide modified protein. Carcinogenesis, 1986, 7(3), 441-444.
[http://dx.doi.org/10.1093/carcin/7.3.441] [PMID: 3948329]
[20]
Casale, G.P.; Rogan, E.G.; Stack, D.; Devanesan, P.; Cavalieri, E.L. Production of a high-affinity monoclonal antibody specific for 7-(benzo[alpha]pyren-6-yl) guanine and its application in a competitive enzyme-linked immunosorbent assay. Chem. Res. Toxicol., 1996, 9(6), 1037-1043.
[http://dx.doi.org/10.1021/tx950203e] [PMID: 8870993]
[21]
Duhachek, S.D.; Kenseth, J.R.; Casale, G.P.; Small, G.J.; Porter, M.D.; Jankowiak, R. Monoclonal antibody-gold biosensor chips for detection of depurinating carcinogen-DNA adducts by fluorescence line-narrowing spectroscopy. Anal. Chem., 2000, 72(16), 3709-3716.
[http://dx.doi.org/10.1021/ac000472w] [PMID: 10959953]
[22]
Roda, A.; Simoni, P.; Ferri, E.N.; Girotti, S.; lus, A.; Rauch, P.; Poplstein, M.; Pospisil, M.; Pipek, P.; Hochel, I.; Fukal, L. Determination of PAHs in various smoked meat products and different samples by enzyme immunoassay. J. Sci. Food Agric., 1999, 79(1), 58-62.
[http://dx.doi.org/10.1002/(SICI)1097-0010(199901)79:1<58::AIDJSFA174>3.0.CO;2-0]
[23]
Zhang, Y.F.; Gao, Z.X. Antibody development and immunoassays for polycyclic aromatic hydrocarbons (PAHs). Curr. Org. Chem., 2017, 21, 2612-2621.
[24]
Chung, M.K.; Riby, J.; Li, H.; Iavarone, A.T.; Williams, E.R.; Zheng, Y.; Rappaport, S.M. A sandwich enzyme-linked immunosorbent assay for adducts of polycyclic aromatic hydrocarbons with human serum albumin. Anal. Biochem., 2010, 400(1), 123-129.
[http://dx.doi.org/10.1016/j.ab.2010.01.018] [PMID: 20083082]
[25]
Russell, A.J.; Trudel, L.J.; Skipper, P.L.; Groopman, J.D.; Tannenbaum, S.R.; Klibanov, A.M. Antibody-antigen binding in organic solvents. Biochem. Biophys. Res. Commun., 1989, 158(1), 80-85.
[http://dx.doi.org/10.1016/S0006-291X(89)80179-2] [PMID: 2912456]
[26]
Setford, S.J.; Kröger, S.; Turner, A.P.F. Organic phase immunosensors. Analusis, 1999, 27(7), 600-608.
[http://dx.doi.org/10.1051/analusis:1999270600]
[27]
Segura-Gil, I.; Blázquez-Soro, A.; Galán-Malo, P.; Mata, L.; Tobajas, A.P.; Sánchez, L.; Pérez, M.D. Development of sandwich and competitive ELISA formats to determine β-conglycinin: Evaluation of their performance to detect soy in processed food. Food Control, 2019, 103, 78-85.
[http://dx.doi.org/10.1016/j.foodcont.2019.03.035]
[28]
Beyer, K.; Knopp, D.; Niessner, R. Problems in immunoassay development of small and volatile molecules-benzene, toluene, and xylenes (BTX). Food Technol. Biotechnol., 1997, 35, 215-223.
[29]
Ecker, C.; Cichna-Markl, M. Development and validation of a sandwich ELISA for the determination of potentially allergenic lupine in food. Food Chem., 2012, 130(3), 759-766.
[http://dx.doi.org/10.1016/j.foodchem.2011.07.100]

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