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標題: 建立奈米磁珠側層流免疫分析法以檢測食品中花生主要過敏原Ara h 1
Develop a magnetic nanoparticle labeled lateral flow assay for the detection of a major peanut allergen Ara h 1 in food.
作者: 李亦婷
Yi-Ting Li
關鍵字: 花生過敏;Ara h 1;免疫磁珠;側層流免疫分析法;peanut allergy;Ara h 1;magnetic nanoparticle;lateral flow assay
引用: 1. Pires F, Arcos- Martinez MJ, Dias-Cabral AC, Vidal JC, Castillo JR. (2018). A rapid magnetic particle-based enzyme immunoassay for humancytomegalovirus glycoprotein B quantification. J Pharm Biomed Anal. 156:372-8. 2. Wittea K, Müllerb K, Grüttner C, Westphal F, Johansson C. (2017). Particle size- and concentration-dependent separation of magnetic nanoparticles. ‎J. Magn. Magn. Mater. 427:320–4. 3. Cho IH, Irudayaraj J. (2013) Lateral-flow enzyme immunoconcentration for rapid detection of Listeria monocytogenes. Anal Bioanal Chem. 405:3313–9. 4. Liu FM, Zhang HL, Wu ZH, Dong HD, Zhou L, Yang DW, Ge YQ, Jia CP, Liu HY, Jin QH, Zhao JL, Zhang QQ, Mao HJ. (2016). Highly sensitive and selective lateral flow immunoassay based on magnetic nanoparticles for quantitative detection of carcinoembryonic antigen. Tanlata. 161: 205-10. 5. Cao M, Li ZH, Wang JL, Ge WP, Yue TL, Li RH, Colvin VL, Yu WW. (2012). ood related applications of magnetic iron oxide nanoparticles: Enzyme immobilization, protein purification, and food analysis. Trends Food Sci. Technol. 27:47-56 6. Song F, Zhou Y, Li YS, Meng XM, Liu JQ, Lu SY, Ren HL, Hu P, Liu ZS, Zhang YY, Zhang JH. (2014). A rapid immunomagnetic beads-based immunoassay for the detection of β-casein in bovine milk. Food chem. 158:445-8. 7. Montiel VRM, Campuzano S, Conzuelo F, Torrente-Rodríguez RM, Gamella M, Reviejo AJ, Pingarrón JM. (2015). Electrochemical magnetoimmunosensing platform for determination of the milk allergen β-lactoglobulin. Tanlata. 131:156-62. 8. Montiel VRM, Torrente-Rodríguez RM, de Rivera GG, Reviejo AJ, Cuadradoc C, Linacero R, Gallego FJ, Campuzano S, Pingarrón JM. (2017). Amperometric determination of hazelnut traces by means of ExpressPCR coupled to magnetic beads assembled on disposable DNA sensingscaffolds. Sens Actuators B Chem. 245: 895-902. 9. Chu PT, Wen HW. (2013). Sensitive detection and quantification of gliadin contamination in gluten-free food with immunomagnetic beads based liposomal fluorescence immunoassay. Anal Chim Acta. 787: 246-253. 10. Zheng C, Wang XC, Lu Y, Liu Y. (2012). Rapid detection of fish major allergen parvalbumin using superparamagnetic nanoparticle-based lateral flow immunoassay. Food Chem. 26:446-52. 11. Careri M, Elviri L, Lagos JB, Mangia A, Speroni F, Terenghi M. (2008). Selective and rapid immunomagnetic bead-based sample treatment for the liquid chromatography–electrospray ion-trap mass spectrometry detection of Ara h3/4 peanut protein in foods. J Chromatogr A. 1206:89-94. 12. Feng XL, Ren HL, Li YS, Hu P, Zhou Y, Liu ZS, Yan DM, Hui Q, Lin C, Liu NN, Liu YY, Lu SY. (2014). A magnetic particles-based chemiluminescence enzyme immunoassay for rapid detection of ovalbumin. Anal Biochem. 459:12-7. 13. Montiel VRM, Campuzano S, Pellicanò A, Torrente-Rodríguez, RM Reviejo AJ, Cosio MS, Pingarrón JM. (2015). Sensitive and selective magnetoimmunosensing platform for determination of the food allergen Ara h 1. Anal Chim Acta. 880:52-9. 14. Zhang YX, Wu QP, Sun M, Zhang JM, Mo SP, Wang JA, Wei XH, B JL. (2018). Magnetic-assisted aptamer-based fluorescent assay for allergendetection in food matrix. Sens Actuators B Chem. 263:43-9. 15. Montiel VRM, Pellicanò A, Campuzano S, Torrente-Rodríguez RM, Reviejo AJ, Cosio MS, Pingarrón JM. (2016) Electrochemical detection of peanuts at trace levels in foods using amagnetoimmunosensor for the allergenic protein Ara h 2. Sens Actuators B Chem.236:825-33. 16. Baumert JL, Tran DH. (2015). Lateral flow devices for detecting allergens in food. Handbook of Food Allergen Detection and Control. Chapter 11. 17. Connolly R, Kennedy RO. (2017). Magnetic lateral flow immunoassay test strip development – Considerations for proof of concept evaluation. Methods. 116:132-40. 18. Liu DF, Huang YM, Wang SY, Liu K, Chen MH, Xiong YH, Yang WC, Lai WH. (2015). A modified lateral flow immunoassay for the detection of trace aflatoxin M1 based on immunomagnetic nanobeads with different antibody concentrations. Food Control. 51:218-24. 19. Yin HY, Chu PT, Tsai WC, Wen HW. (2016). Development of a barcode-style lateral flow immunoassay for the rapid semi-quantification of gliadin in foods. Food Chem. 192:934-42. 20. Zhang XY, Yu XZ, Wen K, Li CL, Mari GM, Jiang HY, Shi WM, Shem JZ, Wang ZH. (2017). Multiplex Lateral Flow Immunoassays Based on Amorphous Carbon Nanoparticles for Detecting Three Fusarium Mycotoxins in Maize. J. agric. food chem. 65:8063-71. 21. Jones KD. (1999). Troubleshooting protein binding in nitrocellulose membranes Part 1: Principles. IVD Technology, 5, 32-41. 22. Masiri J, Barrios-Lopez B, Benoit L, Tamayo J, Day J, Nadala C, Sung SL, Samadpour M. (2016). Development and Validation of a Lateral Flow Immunoassay Test Kit for Dual Detection of Casein and β-Lactoglobulin Residues. J Food Prot. 79(3):477-83. 23. Koizumi D, Shirota K, Akita R, Oda H, Akiyama H. (2014). Development and validation of a lateral flow assay for the detection of crustacean protein in processed foods. Food Chem. 150:348-52. 24. Ji KM, Chen JJ, Gao C, Liu XY, Xia LX, Liu ZG, Li L, Yang SH. (2011). A two-site monoclonal antibody immunochromatography assay for rapid detection of peanut allergen Ara h1 in Chinese imported and exported foods. Food Chem. 129:541-5. 25. Wang Y, Deng R, Zhang G, Li Q, Yang J, Sun Y, Li Z, Hu X. (2015). Rapid and sensitive detection of the food allergen glycinin in powdered milk using a lateral flow colloidal gold immunoassay strip test. J Agric Food Chem. 63(8):2172-8 26. Dunlop JH, Keet CA. (2018). Epidemiology of Food Allergy. Immunol Allergy Clin North Am. 38(1):13-25. 27. Lin YT, Wu CTC, Cheng JH, Hyang JL, Yeh KW. (2012). Patterns of sensitization to peanut allergen components in Taiwanese Preschool children. J Microbiol Immunol Infect. 45,90-5. 28. Lee AJ, Thalayasingam M, Lee BW. (2013). Food allergy in Asia: how does it compare? Asia Pac Allergy. 3:3-14. 29. Osborne NJ, Koplin JJ, Martin PE, Gurrin LC, Lowe AJ, Matheson MC, Ponsonby AL, Wake M, Tang ML, Dharmage SC, Allen KJ; HealthNuts Investigators. (2011). Prevalence of challenge-proven IgE-mediated food allergy using population-based sampling and predetermined challenge criteria in infants. J Allergy Clin Immunol. 127(3):668-76. 30. Waserman S, Watson W. (2011). Food allergy. Allergy Asthma Clin Immunol. 7(Suppl 1):S7 31. Sicherer SH, Wood, RA. (2013). Advances in Diagnosing Peanut Allergy. J Allergy Clin Immunol. 1(1):1-13. 32. Holzhauser T, Röder M. (2011). Allergens in Tree Nuts, Sesame Seeds, Mustard, and Celery. Food Allergens: Analysis Instrumentation and Methods. Chapter 4. 33. Yeung J, Robert MC. (2018). Challenges and Path Forward on Mandatory Allergen Labeling and Voluntary Precautionary Allergen Labeling for a Global Company. ‎J. AOAC Int. 101: 70-6. 34. Allergen Bureau. (2016). The VITAL Program, 35. Ebisawa M, Ito K, Fujisawa T, on behalf of Committee for Japanese Pediatric Guideline for Food Allergy, The Japanese Society of Pediatric Allergy and Clinical Immunology, The Japanese Society of Allergology. (2017). Japanese guidelines for food allergy 2017. Allergol Int. 66:248-64. 36. Leickly FE, Kloepfer KM, Slaven JE, Vitalpur G. (2017). Peanut Allergy: An Epidemiologic Analysis of a Large Database. J Pediatr. 192:223-8. 37. Fæste CK. (2011). Allergens in Peanut, Soybean, and Lupin. Food Allergens: Analysis Instrumentation and Methods. Chapter 3. 38. Hourihane JO, Allen KJ, Shreffler WG, Dunngalvin G, Nordlee JA, Zurzolo GA, Dunngalvin A, Gurrin LC, Baumert JL, Taylor SL. (2017). Peanut Allergen Threshold Study (PATS): Novel single-dose oral food challenge study to validate eliciting doses in children with peanut allergy. J Allergy Clin Immunol. 139(5):1583-90. 39. Uotila R, Kukkonen AK, Blom WM, Remington B, Westerhout J, Pelkonen AS, Mäkelä MJ. (2018). Component-resolved diagnostics demonstrates that most peanut-allergic individuals could potentially introduce tree nuts to their diet. Clin Exp Allergy. 48(6):712-721. 40. Ben-Shoshan M, Harrington DW, Soller L, Fragapane J, Joseph L, St Pierre Y, Godefroy SB, Elliott SJ, Clarke AE. (2010). A population-based study on peanut, tree nut, fish, shellfish, and sesame allergy prevalence in Canada. J Allergy Clin Immunol. 125(6):1327-35. 41. Kanny G, Moneret-Vautrin DA, Flabbee J, Beaudouin E, Morisset M, Thevenin. (2001). Population study of food allergy in France. J Allergy Clin Immunol. 108(1):133-40. 42. Venter C, Hasan Arshad S, Grundy J, Pereira B, Bernie Clayton C, Voigt K, Higgins B, Dean T. (2010). Time trends in the prevalence of peanut allergy: three cohorts of children from the same geographical location in the UK. Allergy. 65(1):103-8. 43. Peng J, Song S, Xu L, Ma W, Liu L, Kuang H, Xu C. (2013). Development of a monoclonal antibody-based sandwich ELISA for peanut allergen Ara h 1 in food. Int J Environ Res Public Health. 10(7):2897-905. 44. Burks AW, Shin D, Cockrell G, Stanley JS, Helm RM, Bannon GA. (1997). Mapping and mutational analysis of the IgE-binding epitopes on Ara h 1, a legume vicilin protein and a major allergen in peanut hypersensitivity. Eur J Biochem. 245(2):334-9. 45. Cabanos C, Urabe H, Tandang-Silvas MR, Utsumi S, Mikami B, Maruyama N. (2011). Crystal structure of the major peanut allergen Ara h 1. Mol Immunol. 49(1-2):115-23 46. Maleki SJ, Chung SY, Champagne ET, Raufman JP. (2000). The effects of roasting on the allergenic properties of peanut proteins. J Allergy Clin Immunol. 106(4):763-8. 47. Beyer K, Morrow E, Li XM, Bardina L, Bannon GA, Burks AW, Sampson HA. (2001). Effects of cooking methods on peanut allergenicity. J Allergy Clin Immunol. 107(6):1077-81. 48. Palmer GW, Dibbern DA Jr, Burks AW, Bannon GA, Bock SA, Porterfield HS, McDermott RA, Dreskin SC. (2005). Comparative potency of Ara h 1 and Ara h 2 in immunochemical and functional assays of allergenicity. Clin Immunol. 115(3):302-12. 49. Jayasena S, Smits M, Fiechter D, de Jong A, Nordlee J, Baumert J, Taylor SL, Pieters RH, Koppelman SJ. (2015). Comparison of six commercial ELISA kits for their specificity and sensitivity in detecting different major peanut allergens. J Agric Food Chem. 63(6):1849-55. 50. Koppelman SJ, Knol EF, Vlooswijk RA, Wensing M, Knulst AC, Hefle SL, Gruppen H, Piersma S. (2003). Peanut allergen Ara h 3: isolation from peanuts and biochemical characterization. Allergy. 58(11):1144-51. 51. Restani P, Ballabio C, Corsini E, Fiocchi A, Isoardi P, Magni C, Poiesi C, Terracciano L, Duranti M. (2005). Identification of the basic subunit of Ara h 3 as the major allergen in a group of children allergic to peanuts. Ann Allergy Asthma Immunol. 94(2):262-6. 52. Wang Y, Fu TJ, Howard A, Kothary MH, McHugh TH, Zhang Y. (2013). Crystal structure of peanut (Arachis hypogaea) allergen Ara h 5. J Agric Food Chem. 61(7):1573-8. 53. Koid AE, Chapman MD, Hamilton RG, van Ree R, Versteeg SA, Dreskin SC, Koppelman SJ, Wünschmann S. (2014). Ara h 6 complements Ara h 2 as an important marker for IgE reactivity to peanut. J Agric Food Chem. 62(1):206-13. 54. Blankestijn MA, Otten HG, Suer W, Weimann A, Knol EF, Knulst AC. (2018). Specific IgE to peanut 2S albumin Ara h 7 has a discriminative ability comparable to Ara h 2 and 6. Clin Exp Allergy. 48(1):60-65. 55. Mittag D, Akkerdaas J, Ballmer-Weber BK, Vogel L, Wensing M, Becker WM, Koppelman SJ, Knulst AC, Helbling A, Hefle SL, Van Ree R, Vieths S. (2004). Ara h 8, a Bet v 1-homologous allergen from peanut, is a major allergen in patients with combined birch pollen and peanut allergy. J Allergy Clin Immunol. 114(6):1410-7. 56. Glaumann S, Nilsson C, Johansson SG, Asarnoj A, Wickman M, Borres MP, Nopp A. (2015). Evaluation of basophil allergen threshold sensitivity (CD-sens) to peanut and Ara h 8 in children IgE-sensitized to Ara h 8. Clin Mol Allergy. 13(1):5. 57. Arkwright PD, Summers CW, Riley BJ, Alsediq N, Pumphrey RS. (2013). IgE sensitization to the nonspecific lipid-transfer protein Ara h 9 and peanut-associated bronchospasm. Biomed Res Int. 746507. 58. Krause S, Reese G, Randow S, Zennaro D, Quaratino D, Palazzo P, Ciardiello MA, Petersen A, Becker WM, Mari A. (2009). Lipid transfer protein (Ara h 9) as a new peanut allergen relevant for a Mediterranean allergic population. J Allergy Clin Immunol. 124(4):771-8. 59. Schwager C, Kull S, Behrends J, Röckendorf N, Schocker F, Frey A, Homann A, Becker WM, Jappe U. (2017). Peanut oleosins associated with severe peanut allergy-importance of lipophilic allergens for comprehensive allergy diagnostics. J Allergy Clin Immunol. 140(5):1331-8 60. Petersen A, Kull S, Rennert S, Becker WM, Krause S, Ernst M, Gutsmann T, Bauer J, Lindner B, Jappe U.(2015). Peanut defensins: Novel allergens isolated from lipophilic peanut extract. J Allergy Clin Immunol. 136(5):1295-301. 61. Bublin M, Breiteneder H. (2014). Cross-Reactivity of Peanut Allergens. Curr Allergy Asthma Rep. 14(4): 426. 62. Cabanillas B, Cuadrado C, Rodriguez J, Hart J, Burbano C, Crespo JF, Novak N. (2015). Potential changes in the allergenicity of three forms of peanut after thermal processing. Food Chem. 183:18-25. 63. Nesbit JB, Hurlburt BK, Schein CH, Cheng H, Wei H, Maleki SJ. (2012). Ara h 1 structure is retained after roasting and is important for enhanced binding to IgE. Mol Nutr Food Res. 56(11):1739-47. 64. Sheu SC, Tsou PC, Lien YY, Lee MS. (2018). Development of loop-mediated isothermal amplification (LAMP) assays for the rapid detection of allergic peanut in processed food. Food Chem. 257:67-74. 65. Yuan D, Kong J, Li X, Fang X, Chen Q. (2018). Colorimetric LAMP microfluidic chip for detecting three allergens: peanut, sesame and soybean. Sci Rep. 8(1):8682. 66. Zhang WJ, Cai Q, Guan X, Chen Q. (2015). Detection of peanut (Arachis hypogaea) allergen by Real-time PCR method with internal amplification control. Food Chem. 174:547-52. 67. Sun X, Jia M, Guan L, Ji J, Zhang Y, Tang L, Li Z. (2015). Multilayer graphene-gold nanocomposite modified stem-loop DNA biosensor for peanut allergen-Ara h1 detection. Food Chem. 172:335-42 68. Weng X, Neethirajan S. (2016). A microfluidic biosensor using graphene oxide and aptamer-functionalized quantum dots for peanut allergen detection. Biosens Bioelectron. 85:649-656 69. Thomas H, Kornelia K, Annabella J, Martin R. (2014). Matrix-normalised quantification of species by threshold-calibrated competitive real-time PCR Allergenic peanut in food as one example. Food Chem. 163:68–76. 70. Mustorp SL, Drømtorp SM, Holck AL. (2011). Multiplex, quantitative, ligation-dependent probe amplification for determination of allergens in food. J Agric Food Chem. 59(10):5231-9. 71. Stephan O, Vieths S. (2004). Development of a real-time PCR and a sandwich ELISA for detection of potentially allergenic trace amounts of peanut (Arachis hypogaea) in processed foods. J Agric Food Chem. 52(12):3754-60. 72. Alves RC, Pimentel FB, Nouws HP, Marques RC, González-García MB, Oliveira MB, Delerue-Matos C. (2015). Detection of Ara h 1 (a major peanut allergen) in food using an electrochemical gold nanoparticle-coated screen-printed immunosensor. Biosens Bioelectron. 64:19-24. 73. Chen H, Zou Z, Tao A. (2018). A Quantitative Method for Detecting Ara h 2 by Generation and Utilization of Monoclonal Antibodies. J Immunol Res. 2018:4894705. 74. Pomés A, Vinton R, Chapman MD. (2004). Peanut allergen (Ara h 1) detection in foods containing chocolate. J Food Prot. 67(4):793-8. 75. Pomés A, Helm RM, Bannon GA, Burks AW, Tsay A, Chapman MD. (2003). Monitoring peanut allergen in food products by measuring Ara h 1. J Allergy Clin Immunol. 111(3):640-5. 76. Speroni F, Elviri L, Careri M, Mangia A. (2010). Magnetic particles functionalized with PAMAM-dendrimers and antibodies: a new system for an ELISA method able to detect Ara h3/4 peanut allergen in foods. Anal Bioanal Chem. 397(7):3035-42. 77. Juan P, Shanshan S, Liqiang L, Hua K, Chuanlai X. (2015). Development of Sandwich ELISA and Immunochromatographic Strip for the Detection of Peanut Allergen Ara h 2. Food Anal. Methods, 8:2605–2611. 78. Wen HW, Borejsza-Wysocki W, DeCory TR, Durst RA. (2005). Development of a competitive liposome-based lateral flow assay for the rapid detection of the allergenic peanut protein Ara h1. Anal Bioanal Chem. 382(5):1217-26. 79. Costa J, Mafra I, Carrapatoso I, Oliveira MB. (2012). Almond allergens: molecular characterization, detection, and clinical relevance. J Agric Food Chem. 60(6):1337-49. 80. Spremulli, L. (2000). Protein synthesis, assembly and degradation. In: B. B. Buchanan, W. Gruissem, & R. L. Jones (Eds.), Biochemistry& molecular biology of plants (pp. 412-454). American Society of Plant Physiologists, Rockville, MD, USA. 81. Schneider, Z. (1980). Aliphatic alcohols improve the adsorptive performance of cellulose nitrate membranes-application in chromatography and enzyme assays. Anal Biochem, 108, 96-103. 82. Bagherpour AR, Kashanian F, Seyyed Ebrahimi SA, Habibi-Rezaei M. (2018). L-arginine modified magnetic nanoparticles: green synthesis and characterization. Nanotechnology. 29(7):075706. 83. Pollet J, Delport F, Janssen KP, Tran DT, Wouters J, Verbiest T, Lammertyn J. (2011). Fast and accurate peanut allergen detection with nanobead enhanced optical fiber SPR biosensor. Talanta. 83(5):1436-41. 84. Walczyk NE, Smith PMC, Tovey ER, Roberts TH. (2017). Peanut protein extraction conditions strongly influence yield of allergens Ara h 1 and 2 and sensitivity of immunoassays. Food Chem. 221:335-344. 85. Koppelman SJ, Bruijnzeel-Koomen CA, Hessing M, de Jongh HH. (1999). Heat-induced conformational changes of Ara h 1, a major peanut allergen, do not affect its allergenic properties. J Biol Chem. 274(8):4770-7. 86. Venosa, A. D., King, D. W. & Sorial, G. A. (2002). The baffled flask test for dispersant effectiveness: a round robin evaluation of reproducibility and repeatability. Spill Sci. Technol. Bull, 7:299-308. 87. Wingfield PT. (2016). Protein Precipitation Using Ammonium Sulfate. Curr Protoc Protein Sci. 84:A.3F.1-9 88. Ghosh R, Gilda JE, Gomes AV. (2014). The necessity of and strategies for improving confidence in the accuracy of western blots. Expert Rev Proteomics. 11(5):549-60. 89. Engvall E, Jonsson K, Perlmann P. (1971). Enzyme-linked immunosorbent assay. II. Quantitative assay of protein antigen, immunoglobulin G, by means of enzyme-labelled antigen and antibody-coated tubes. Biochim Biophys Acta. 251(3):427-34. 90. Tzen JT, Peng CC, Cheng DJ, Chen EC, Chiu JM. (1997). A new method for seed oil body purification and examination of oil body integrity following germination. J Biochem. 121(4):762-8.
花生過敏可能導致高危險的急性症狀發生,僅微量的花生過敏原就能引起嚴重的過敏反應,且可能藉由加工過程交互汙染,因此需要建立一個快速偵測花生過敏原的系統去預防以及產品品質管控。側層流免疫分析法做為一個便利且肉眼可視的檢測系統已廣泛應用在各種領域上,結合免疫磁珠分離法可以有效增加靈敏度。Ara h 1為花生的主要過敏原,且在熱處理的過程中可能增加致敏性,因此以Ara h 1做為系統的偵測對象。選擇Anti-Ara h 1全蛋白抗體及一株核心區域胜肽抗體配對後,針對試紙與磁珠進行條件最適化。試紙與磁珠的抗體皆為Anti-Ara h 1全蛋白抗體,試紙AE99以glycine做為blocker、檢量線抗體密度為2.5 μg/cm;磁珠150 nm以750 μg/ml抗體修飾,並以2.5%脫脂牛奶blocking;磁珠樣品比例為20:300。以此模式在溶液中的肉眼可視極限為0.01 μg/ml,標準方程式y = 30.967ln(x) + 166.6,R² = 0.9914。針對進行巧克力、餅乾及牛奶三種基質進行測試,餅乾與牛奶檢測極限為0.5 μg/ml而巧克力為1 μg/ml,並在食品樣品中可正確檢測到含有花生的產品。

Peanut can induce severe allergic reaction by very small amount. In the food processing, peanut allergen residues may contaminate other products which do not contain peanut, it will be an important problem to peanut-allergic consumers. In order to prevent peanut allergen from contamination and improve the quality control, an analytical method for rapid detecting peanut allergen is necessary. Lateral flow assay (LFA) is known for its simple procedure to detect target rapidly. Magnetic nanoparticles can against matrix effect and improve detection result. As a peanut major allergen, Ara h 1, may increase allergenicity after heat processing. Therefore, in this study, a LFA model based on magnetic nanoparticle for the rapid detection of Ara h 1 was developed. Firstly, anti-Ara h 1 polyclonal antibody was selected to be tagged on magnetic nanoparticles and immobilized on the test line of strip. Following was the optimal conditions: Nitrocellulose membrane type was AE99; antibody density on the test line is 2.5 μg/cm; blocker on membrane was glycine; magnetic nanoparticle size was 150 nm modified antibody concentration is 750 μg/ml; magnetic beads blocker was non-fat dried milk; ratio of magnetic beads and sample is 20:300 (v:v). The visual limit of detection is 0.01 μg/ml Ara h 1 in PBS, and the formula was y = 30.967ln(x) + 166.6 with R square of 0.9914. This assay was used to detect peanut allergen-Ara h 1 in cookie, milk and chocolate product, and the detection limit was 0.5 μg/ml (cookie, milk) and 1μg/ml (chocolate). Moreover, Ara h 1 in various food smples were detected by the developed assay.
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