臺灣博碩士論文加值系統

花生 (Arachis hypogaea) 為一營養豐富且大眾化的食品，但據估計全世界約有1%至2%的人口對花生過敏，患者若不慎誤食花生可能會引發嚴重的過敏反應，而花生中主要的過敏原為Ara h 1、Ara h 2及Ara h 3。現今常見用來檢測過敏原的方式為酵素連結免疫吸附分析法 (Enzyme-linked immunosorbent assay, ELISA)，然而目前市面上相關檢測套組數量少，且價格相當昂貴，因此本研究目標為開發可用於檢測花生過敏原之三明治型ELISA。本研究首先從花生中萃取花生蛋白，以硫酸銨鹽析初步純化，並使用膠體過濾層析提高純化蛋白質之純度，接著以紐西蘭白兔進行免疫生產多株抗體，再利用免疫親和層析法純化血清中的抗體，以山葵過氧化酶 (Horseradish peroxidase, HRP) 標記後作為偵測抗體，並以此建立檢測花生過敏原之三明治型ELISA，最後進行一系列確效分析測試。實驗結果顯示，Ara h 1、Ara h 2及Ara h 3透過SDS-PAGE得知具有良好的純度，將它們作為免疫原個別以紐西蘭白兔製備抗體，並利用免疫擴散法進行測試，可以發現抗血清具有良好的效價。接著將純化後之Ara h 3抗體以HRP標記，透過直接法ELISA測試，結果顯示其可以作為偵測抗體使用，且稀釋之比例以1:20000為最佳。最後將Ara h 3抗體作為捕獲抗體，純化之Ara h 3為標準品，HRP標記之Ara h 3抗體作為偵測抗體，建立三明治型ELISA，捕獲抗體濃度為1 μg/mL，標準品濃度範圍為0.024-25 μg/mL，R2 = 0.999。確效分析方面，intra-day及inter-day之變異係數分別為0.84-5.21%及0.87-6.12%，兩者皆低於10%。LOD及LOQ分別為0.023 μg/mL及0.033 μg/mL。而本方法之回收率介於95.5%-107.5%，表示本方法之精密度及準確度良好，且不與腰果、芝麻及黃豆產生交叉反應性。由上述結果可判定，本研究建立的三明治型ELISA可用於檢測花生中過敏原Ara h 3。未來將延伸本ELISA技術模式至Ara h 1及Ara h 2之檢測。

Peanuts (Arachis hypogaea) is a nutritious and popular food, but it is estimated that about 1% to 2% of the world’s population are allergic to peanuts. If the patient accidentally eats peanut, it may cause severe allergic reactions. The major allergens of peanut are Ara h 1, Ara h 2 and Ara h 3. Nowadays, the common method used to detect allergens is Enzyme-linked immunosorbent assay (ELISA). However, the number of relevant test kits currently on the market is small and the price is quite expensive. Therefore, the aim of this study is to develop sandwich ELISA for detecting peanut allergens. Firstly, peanut protein is extracted from peanuts, then initially purified by ammonium sulfate salting out, and purified by gel filtration chromatography to improve the purity of the purified protein. Then, New Zealand white rabbits were selected for immune production of polyclonal antibodies, and the serum is purified by immunoaffinity chromatography. The antibody was labeled with horseradish peroxidase (HRP) and used as a detection antibody to develop a sandwich ELISA for the detection of peanut allergens. Finally, a series of validation analysis tests are performed. The experimental results showed that the purification of Ara h 1, Ara h 2 and Ara h 3 was found that they have good purity through SDS-PAGE. They were used as immunogens to immunize New Zealand white rabbits to prepare antibodies, and tested by immunodiffusion method, it can be found that the antiserum has good titer. Then, the purified Ara h 3 antibody was labeled to HRP and tested by direct ELISA. The experimental results showed that it can be used as a detection antibody, and the dilution ratio of 1:20000 is the best. Finally, the Ara h 3 antibody was used as the capture antibody, the purified Ara h 3 was used as the standard, and the HRP-labeled Ara h 3 antibody was used as the detection antibody to develop a sandwich ELISA. The concentration of the capture antibody was 1 μg/mL, and the concentration range of the standard was 0.024-25 μg/mL, R2 = 0.999. In terms of verification analysis, the coefficients of variation of intra-day and inter-day are 0.84-5.21% and 0.87-6.12%, respectively, both of which are lower than 10%. LOD and LOQ are 0.023 μg/mL and 0.033 μg/mL, respectively, and the recovery of this assay is between 95.5% and 107.5%, indicating that the precision and accuracy of this method is good, and it does not cross-react with cashew nuts, sesame, and soybeans. From the above results, it can be determined that the sandwich ELISA we developed in this study can be used to detect the allergen Ara h 3 in peanuts. In the future, this ELISA technology pattern will be extended to the detection of Ara h 1 and Ara h 2.

摘要 I
Abstract II
謝誌 IV
目錄 V
圖目錄 IX
表目錄 XI
壹 前言 1
貳 文獻回顧 2
一、 食物過敏概述 2
二、 過敏引發機制 2
三、 過敏原命名原則 3
四、 花生過敏原 5
(一) 主要花生過敏原 7
(二) 次要花生過敏原 12
五、 蛋白質純化技術 15
(一) 概述 15
(二) 沉澱法 16
(三) 分子排阻層析法 (Size exclusion chromatography, SEC) 21
(四) 離子交換層析 (Ion exchange chromatography) 21
(五) 親和性層析 22
六、 十二烷基硫酸鈉聚丙烯醯胺凝膠電泳 (Sodium dodecyl sulfate polyacrylamide gel electrophoresis, SDS-PAGE) 22
七、 西方墨點法 (Western blot) 22
八、 免疫擴散法 (Immunodiffusion assay) 23
九、 過敏原檢測方法 25
(一) 酵素連結免疫吸附分析法 (Enzyme-linked immunosorbent assay, ELISA) 25
(二) 聚合酶連鎖反應 (Polymerase chain reaction techniques, PCR) 28
(三) 即時聚合酶連鎖反應 (Real- time PCR) 28
(四) 生物傳感器檢測技術 30
十、 抗體 (Antibody, Ab) 30
(一) 多株抗體 (Polyclonal antibody) 33
(二) 單株抗體 (Monoclonal antibody) 33
十一、 佐劑 34
參 材料與方法 35
一、 實驗架構設計 35
二、 實驗材料與儀器 36
(一) 實驗樣品 36
(二) 實驗藥品 36
(三) 實驗設備 38
三、 實驗方法 40
(一) 花生樣品預處理 40
(二) 花生蛋白萃取 40
(三) 硫酸銨分級沉澱 (Ammonium sulfate fractionation, ASF) 40
(四) 膠體過濾層析 40
(五) 花生過敏原分析 41
(六) 可溶性蛋白含量測定 43
(七) 動物免疫試驗 43
(八) 兔抗血清中Ara h 3抗體純化 45
(九) Ara h 3抗體之標記 46
(十) Direct ELISA測試 46
(十一) Sandwich ELISA建立 47
(十二) 確效分析 47
肆 結果與討論 49
一、 硫酸銨分級沉澱 49
(一) Ara h 1之純化 51
(二) Ara h 2之純化 51
(三) Ara h 3之純化 51
二、 花生過敏原分析 56
三、 抗體製備 59
(一) Rabbit anti-Ara h 1抗體檢測 59
(二) Rabbit anti-Ara h 2抗體檢測 59
(三) Rabbit anti-Ara h 3抗體檢測 59
四、 Rabbit Anti-Ara h 3 抗體純化 63
五、 建立檢測Ara h 3之ELISA 65
(一) Direct ELISA測試 65
(二) Sandwich ELISA建立 68
(三) 確效分析 70
六、 市售產品應用 71
伍 結論 75
陸 參考文獻 76
附錄一 90
作者簡介 91

衛生福利部食品藥物管理署 (2018) 實驗室品質管理規範－化學領域測試結果之品質管制。台北市：行政院。

Abbott, M., Hayward, S., Ross, W., Godefroy, S. B., Ulberth, F., Van Hengel, A. J., ... & Delahaut, P. (2010). Validation procedures for quantitative food allergen ELISA methods: community guidance and best practices. Journal of AOAC International, 93(2), 442-450.
Alhajj, M., & Farhana, A. (2021). Enzyme Linked Immunosorbent Assay. StatPearls.
Ansar, W., & Ghosh, S. (2013). Monoclonal antibodies: a tool in clinical research. Indian Journal of Clinical Medicine, 4, IJCM-S11968.
Anzengruber, J., Bublin, M., Bönisch, E., Janesch, B., Tscheppe, A., Braun, M. L., ... & Schäffer, C. (2017). Lactobacillus buchneri S-layer as carrier for an Ara h 2-derived peptide for peanut allergen-specific immunotherapy. Molecular immunology, 85, 81-88.
Aydin, S. (2015). A short history, principles, and types of ELISA, and our laboratory experience with peptide/protein analyses using ELISA. Peptides, 72, 4-15.
Bailey, G. S. (1996). Ouchterlony Double Immunodiffusion. The Protein Protocols Handbook, 749-752.
Barre, A., Simplicien, M., Cassan, G., Benoist, H., & Rougé, P. (2018). Oil bodies (oleosomes): Occurrence, structure, allergenicity. Revue Française d'Allergologie, 58(8), 574-580.
Becker, W. M., & Jappe, U. (2014). Peanut allergens. In History of Allergy (Vol. 100, pp. 256-267). Karger Publishers.
Bernard, H., Guillon, B., Drumare, M. F., Paty, E., Dreskin, S. C., Wal, J. M., ... & Hazebrouck, S. (2015). Allergenicity of peanut component Ara h 2: Contribution of conformational versus linear hydroxyproline-containing epitopes. Journal of Allergy and Clinical Immunology, 135(5), 1267-1274.
Brooijmans, T., Breuer, P., Schoenmakers, P. J., & Peters, R. A. (2020). Charge-based separation of synthetic macromolecules by non-aqueous ion exchange chromatography. Journal of Chromatography A, 1626, 461351.
Brunelle, J. L., & Green, R. (2014). One-dimensional SDS-polyacrylamide gel electrophoresis (1D SDS-PAGE). Methods in enzymology, 541, 151-159.
Brusotti, G., Calleri, E., Colombo, R., Massolini, G., Rinaldi, F., & Temporini, C. (2018). Advances on size exclusion chromatography and applications on the analysis of protein biopharmaceuticals and protein aggregates: a mini review. Chromatographia, 81(1), 3-23.
Bublin, M., & Breiteneder, H. (2014). Cross-reactivity of peanut allergens. Current allergy and asthma reports, 14(4), 1-12.
Bublin, M., Kostadinova, M., Radauer, C., Hafner, C., Szépfalusi, Z., Varga, E. M., ... & Breiteneder, H. (2013). IgE cross-reactivity between the major peanut allergen Ara h 2 and the nonhomologous allergens Ara h 1 and Ara h 3. J. Allergy Clin. Immunol. 132: 118-124.
Burgess, R. R. (2009). Protein precipitation techniques. Methods in enzymology, 463, 331-342.
Burgess, R. R. (2018). A brief practical review of size exclusion chromatography: rules of thumb, limitations, and troubleshooting. Protein expression and purification, 150, 81-85.
Burks, A. W., Shin, D., Cockrell, G., Stanley, J. S., Helm, R. M., & Bannon, G. A. (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. European journal of biochemistry, 245(2), 334-339.
Burks, A. W. (2008). Peanut allergy. The Lancet, 371(9623), 1538-1546.
Buss, N. A., Henderson, S. J., McFarlane, M., Shenton, J. M., & De Haan, L. (2012). Monoclonal antibody therapeutics: history and future. Current opinion in pharmacology, 12(5), 615-622.
Cabanos, C., Tandang-Silvas, M. R., Odijk, V., Brostedt, P., Tanaka, A., Utsumi, S., & Maruyama, N. (2010). Expression, purification, cross-reactivity and homology modeling of peanut profilin. Protein expression and purification, 73(1), 36-45.
Chandel, K. K., & Pahadiya, S. (2005). Sodium Dodecyl Sulfate-Poly Acrylamide Gel Electrophoresis.
Chapman, M. D., Pomés, A., Breiteneder, H., & Ferreira, F. (2007). Nomenclature and structural biology of allergens. Journal of allergy and clinical immunology, 119(2), 414-420.
Chen, W., Cheng, T. N., Khaw, L. F., Li, X., Yang, H., Ouyang, J., & Heng, J. Y. (2021). Protein purification with nanoparticle-enhanced crystallisation. Separation and Purification Technology, 255, 117384.
Chen, X., Negi, S. S., Liao, S., Gao, V., Braun, W., & Dreskin, S. C. (2016). Conformational IgE epitopes of peanut allergens Ara h 2 and Ara h 6. Clin. Exp. Allergy. 46: 1120–1128.
Cooper, H. M., & Patterson, Y. (2008). Production of polyclonal antisera. Current protocols in immunology, 82(1), 2-4.
Costa, J., Ansari, P., Mafra, I., Oliveira, M. B. P., & Baumgartner, S. (2015). Development of a sandwich ELISA-type system for the detection and quantification of hazelnut in model chocolates. Food chemistry, 173, 257-265.
Deng, X., Liu, L., Ma, W., Xu, C., Wang, L., & Kuang, H. (2012). Development and validation of a sandwich ELISA for quantification of peanut agglutinin (PNA) in foods. Food and Agricultural Immunology, 23(3), 265-272.
Duong-Ly, K. C., & Gabelli, S. B. (2014). Salting out of proteins using ammonium sulfate precipitation. Methods in enzymology, 541, 85-94.
Fang, Y. M., Lin, D. Q., & Yao, S. J. (2018). Review on biomimetic affinity chromatography with short peptide ligands and its application to protein purification. Journal of Chromatography A, 1571, 1-15.
Fekete, S., Beck, A., Veuthey, J. L., & Guillarme, D. (2015). Ion-exchange chromatography for the characterization of biopharmaceuticals. Journal of pharmaceutical and biomedical analysis, 113, 43-55.
Gan, S. D., & Patel, K. R. (2013). Enzyme immunoassay and enzyme-linked immunosorbent assay. J Invest Dermatol, 133(9), e12.
Garibyan, L., & Avashia, N. (2013). Research techniques made simple: polymerase chain reaction (PCR). The Journal of investigative dermatology, 133(3), e6.
Grossmann, L., Ebert, S., Hinrichs, J., & Weiss, J. (2018). Effect of precipitation, lyophilization, and organic solvent extraction on preparation of protein-rich powders from the microalgae Chlorella protothecoides. Algal research, 29, 266-276.
Hagel, L. (1998). Gel‐filtration chromatography. Current protocols in molecular biology, 44(1), 10-9.
Hales, B. J., Bosco, A., Mills, K. L., Hazell, L. A., Loh, R., Holt, P. G., & Thomas, W. R. (2004). Isoforms of the major peanut allergen Ara h 2: IgE binding in children with peanut allergy. International archives of allergy and immunology, 135(2), 101-107.
Hanack, K., Messerschmidt, K., & Listek, M. (2016). Antibodies and selection of monoclonal antibodies. Protein Targeting Compounds, 11-22.
Hanna, S. E., Connor, C. J., & Wang, H. H. (2005). Real‐time polymerase chain reaction for the food microbiologist: technologies, applications, and limitations. Journal of Food Science, 70(3), R49-R53.
Hayen, S. M., Ehlers, A. M., den Hartog Jager, C. F., Garssen, J., Knol, E. F., Knulst, A. C., ... & Otten, H. G. (2018). 2S protein Ara h 7.0201 has unique epitopes compared to other Ara h 7 isoforms and is comparable to 2S proteins Ara h 2 and 6 in basophil degranulation capacity. Clinical & Experimental Allergy, 48(7), 890-897.
Hazebrouck, S., Guillon, B., Paty, E., Dreskin, S. C., Adel‐Patient, K., & Bernard, H. (2019). Variable IgE cross‐reactivity between peanut 2S‐albumins: The case for measuring IgE to both Ara h 2 and Ara h 6. Clinical & Experimental Allergy, 49(8), 1107-1115.
Hemmings, O., Du Toit, G., Radulovic, S., Lack, G., & Santos, A. F. (2020). Ara h 2 is the dominant peanut allergen despite similarities with Ara h 6. Journal of Allergy and Clinical Immunology, 146(3), 621-630.
Hnasko, T. S., & Hnasko, R. M. (2015). The western blot. In ELISA (pp. 87-96). Humana Press, New York, NY.
Hornbeck, P. (2017). Double-immunodiffusion assay for detecting specific antibodies (Ouchterlony). Curr. Protoc. Immunol. 116: 2.3.1-2.3.4.
Iqbal, A., Farooq Shah, Y. J., Hamayun, M., & Islam, B. (2018). Detection of food allergens by ELISA and other common methods. FEB-FRESENIUS ENVIRONMENTAL BULLETIN, 8340.
Joo Chan, C., Richardo, T., & Lim, R. L. H. (2018). Current trend in immunotherapy for peanut allergy. International reviews of immunology, 37(6), 279-290.
Joshi, M., & Deshpande, J. D. (2010). Polymerase chain reaction: methods, principles and application. International Journal of Biomedical Research, 2(1), 81-97.
Kang, I. H., & Gallo, M. (2007). Cloning and characterization of a novel peanut allergen Ara h 3 isoform displaying potentially decreased allergenicity. Plant science, 172(2), 345-353.
Kim, B. (2017). Western blot techniques. In Molecular Profiling (pp. 133-139). Humana Press, New York, NY.
Kleber-Janke, T., Crameri, R., Scheurer, S., Vieths, S., & Becker, W. M. (2001). Patient-tailored cloning of allergens by phage display: peanut (Arachis hypogaea) profilin, a food allergen derived from a rare mRNA. Journal of Chromatography B: Biomedical Sciences and Applications, 756(1-2), 295-305.
Koppelman, S. J., Hefle, S. L., Taylor, S. L., & De Jong, G. A. (2010). Digestion of peanut allergens Ara h 1, Ara h 2, Ara h 3, and Ara h 6: a comparative in vitro study and partial characterization of digestion‐resistant peptides. Molecular nutrition & food research, 54(12), 1711-1721.
Koppelman, S. J., Knol, E. F., Vlooswijk, R. A. A., Wensing, M., Knulst, A. C., Hefle, S. L., ... & Piersma, S. (2003). Peanut allergen Ara h 3: isolation from peanuts and biochemical characterization. Allergy, 58(11), 1144-1151.
Krause, S., Reese, G., Randow, S., Zennaro, D., Quaratino, D., Palazzo, P., ... & Mari, A. (2009). Lipid transfer protein (Ara h 9) as a new peanut allergen relevant for a Mediterranean allergic population. Journal of Allergy and Clinical Immunology, 124(4), 771-778.
Lauer, I., Dueringer, N., Pokoj, S., Rehm, S., Zoccatelli, G., Reese, G., ... & Scheurer, S. (2009). The non‐specific lipid transfer protein, Ara h 9, is an important allergen in peanut. Clinical & Experimental Allergy, 39(9), 1427-1437.
Leenaars, M., & Hendriksen, C. F. (2005). Critical steps in the production of polyclonal and monoclonal antibodies: evaluation and recommendations. Ilar Journal, 46(3), 269-279.
Lindblad, E. B. (2000). Freund's adjuvants. In Vaccine Adjuvants (pp. 49-63). Springer, Totowa, NJ.
Liu, J. Q., Hu, T. Y., Diao, K. Y., Yu, D., Song, Y. N., Mo, L. H., Yang, G., Liu, Z. Q., Liu, Z. G., Yang, P. C. (2020). Cold stress promotes IL-33 expression in intestinal epithelial cells to facilitate food allergy development. Cytokine, 136: 155295.
Liu, Y., Fang, X., Sun, X., Niu, B., & Chen, Q. (2021). Detection of Allergen Genes in Peanut and Soybean by Circular Fluorescence Probe-Mediated Isothermal Amplification. Food Analytical Methods, 14(3), 453-464.
Magnusdottir, H., Vidarsdóttir, A. G., Ludviksson, B. R., Clausen, M., Lund, S. H., Jensen, A. B., & Sigurdardottir, S. T. (2019). Ara h 1 and Ara h 6 Sensitization Causes Clinical Peanut Allergy in Ara h 2-Negative Individuals. International archives of allergy and immunology, 178(1), 66-75.
Mahmood, T., & Yang, P. C. (2012). Western blot: technique, theory, and trouble shooting. North American journal of medical sciences, 4(9), 429.
Maleki, S. J., Kopper, R. A., Shin, D. S., Park, C. W., Compadre, C. M., Sampson, H., Burks, A. W. & Bannon, G. A. (2000). Structure of the major peanut allergen Ara h 1 may protect IgE-binding epitopes from degradation. J. Immunol 164: 5844-5849.
Martinet, J., Couderc, L., Renosi, F., Bobée, V., Marguet, C., & Boyer, O. (2016). Diagnostic value of antigen-specific immunoglobulin E immunoassays against Ara h 2 and Ara h 8 peanut components in child food allergy. International archives of allergy and immunology, 169(4), 216-222.
Masuyama, K., Yamamoto, K., Ito, K., Kitagawa, E., & Yamaki, K. (2014). Simplified methods for purification of peanut allergenic proteins: Ara h 1, Ara h 2, and Ara h 3. Food Sci. Technol. Res. 20(4), 875-881.
Matak, K. E., Tahergorabi, R., & Jaczynski, J. (2015). A review: Protein isolates recovered by isoelectric solubilization/precipitation processing from muscle food by-products as a component of nutraceutical foods. Food Research International, 77, 697-703.
Matsuo, H., Yokooji, T., & Taogoshi, T. (2015). Common food allergens and their IgE-binding epitopes. Allergology International, 64(4), 332-343.
Mittag, D., Akkerdaas, J., Ballmer-Weber, B. K., Vogel, L., Wensing, M., Becker, W. M., ... & 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. Journal of Allergy and Clinical Immunology, 114(6), 1410-1417.
Mielenz, M., Mielenz, B., Singh, S. P., Kopp, C., Heinz, J., Häussler, S., & Sauerwein, H. (2013). Development, validation, and pilot application of a semiquantitative Western blot analysis and an ELISA for bovine adiponectin. Domestic animal endocrinology, 44(3), 121-130.
Morais, S., Tortajada-Genaro, L. A., Maquieira, A., & Martinez, M. A. G. (2020). Biosensors for food allergy detection according to specific IgE levels in serum. TrAC Trends Analyt. Chem. 115904.
Nelson, P. N., Reynolds, G. M., Waldron, E. E., Ward, E., Giannopoulos, K., & Murray, P. G. (2000). Demystified…: monoclonal antibodies. Molecular pathology, 53(3), 111.
Novák, P., & Havlíček, V. (2016). Protein extraction and precipitation. In Proteomic profiling and analytical chemistry (pp. 51-62). Elsevier.
Ó’Fágáin, C., Cummins, P. M., & O’Connor, B. F. (2011). Gel-filtration chromatography. Protein Chromatography, 25-33.
Otsu, K., Guo, R., & Dreskin, S. C. (2015). Epitope analysis of Ara h 2 and Ara h 6: characteristic patterns of IgE‐binding fingerprints among individuals with similar clinical histories. Clinical & Experimental Allergy, 45(2), 471-484.
Pan, D., Tang, B., Liu, H., Li, Z., Ma, R., Peng, Y., ... & Wang, Y. (2020). Effect of High Hydrostatic Pressure (HHP) Processing on Immunoreactivity and Spatial Structure of Peanut Major Allergen Ara h 1. Food and Bioprocess Technology, 13(1), 132-144.
Pandey, A. K., Varshney, R. K., Sudini, H. K., & Pandey, M. K. (2019). An Improved Enzyme-Linked Immunosorbent Assay (ELISA) Based Protocol Using Seeds for Detection of Five Major Peanut Allergens Ara h 1, Ara h 2, Ara h 3, Ara h 6, and Ara h 8. Frontiers in nutrition, 6, 68.
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. International Journal of Environmental Research and Public Health, 10(7), 2897-2905.
Pepper, A. N., Assa'ad, A., Blaiss, M., Brown, E., Chinthrajah, S., Ciaccio, C., Fasano, M. B., Gupta, R., Hong, N., Lang, D., Mahr, T., Malawer, E., Roach, A., Shreffler, W., Sicherer, S., Vickers, K., Vickery, B. P., Wasserman, R., Yates, K. & Casale, T. B. (2020). Consensus Report from the Food Allergy Research and Education (FARE) 2019 Oral Immunotherapy for Food Allergy Summit. J. Allergy Clin. Immunol. 146: 244-249
Petersen, A., Kull, S., Rennert, S., Becker, W. M., Krause, S., Ernst, M., ... & Jappe, U. (2015). Peanut defensins: Novel allergens isolated from lipophilic peanut extract. Journal of Allergy and Clinical Immunology, 136(5), 1295-1301.
Pi, X., Wan, Y., Yang, Y., Li, R., Wu, X., Xie, M., Li, X. & Fu, G. (2019). Research progress in peanut allergens and their allergenicity reduction. Trends Food Sci Technol. 93: 212-220.
Pires, I. S., & Palmer, A. F. (2021). Selective protein purification via tangential flow filtration–Exploiting protein-protein complexes to enable size-based separations. J. Membr. Sci. 618: 118712.
Pollet, J., Delport, F., Janssen, K. P. F., Tran, D. T., Wouters, J., Verbiest, T., & Lammertyn, J. (2011). Fast and accurate peanut allergen detection with nanobead enhanced optical fiber SPR biosensor. Talanta, 83(5), 1436-1441.
Pomés, A., Davies, J. M., Gadermaier, G., Hilger, C., Holzhauser, T., Lidholm, J., ... & Goodman, R. E. (2018). WHO/IUIS Allergen Nomenclature: Providing a common language. Molecular immunology, 100, 3-13.
Poms, R. E., Anklam, E., & Kuhn, M. (2004). Polymerase chain reaction techniques for food allergen detection. Journal of AOAC International, 87(6), 1391-1397.
Pons, L., Chéry, C., Mrabet, N., Schohn, H., Lapicque, F., & Guéant, J. L. (2005). Purification and cloning of two high molecular mass isoforms of peanut seed oleosin encoded by cDNAs of equal sizes. Plant Physiology and Biochemistry, 43(7), 659-668.
Powledge, T. M. (2004). The polymerase chain reaction. Advances in physiology education, 28(2), 44-50.
Rabjohn, P., Helm, E. M., Stanley, J. S., West, C. M., Sampson, H. A., Burks, A. W., & Bannon, G. A. (1999). Molecular cloning and epitope analysis of the peanut allergen Ara h 3. The Journal of clinical investigation, 103(4), 535-542.
Rekima, A., Bonnart, C., Macchiaverni, P., Metcalfe, J., Tulic, M. K., Halloin, N., Rekima, S. Genuneit, J. Zanelli, S. Medeiros, S. Palmer, D. J. Prescott, S. &Verhasselt, V. (2020). A role for early oral exposure to house dust mite allergens through breast milk in IgE-mediated food allergy susceptibility. J. Allergy Clin. Immunol. 145: 1416-1429.
Sakamoto, S., Putalun, W., Vimolmangkang, S., Phoolcharoen, W., Shoyama, Y., Tanaka, H., & Morimoto, S. (2018). Enzyme-linked immunosorbent assay for the quantitative/qualitative analysis of plant secondary metabolites. Journal of natural medicines, 72(1), 32-42.
Schwager, C., Kull, S., Behrends, J., Röckendorf, N., Schocker, F., Frey, A., ... & Jappe, U. (2017). Peanut oleosins associated with severe peanut allergy—importance of lipophilic allergens for comprehensive allergy diagnostics. Journal of Allergy and Clinical Immunology, 140(5), 1331-1338.
Shah, K., & Maghsoudlou, P. (2016). Enzyme-linked immunosorbent assay (ELISA): the basics. British journal of hospital medicine, 77(7), C98-C101.
Shah, F., Shi, A., Ashley, J., Kronfel, C., Wang, Q., Maleki, S. J., ... & Zhang, J. (2019). Peanut allergy: Characteristics and approaches for mitigation. Comprehensive reviews in food science and food safety, 18(5), 1361-1387.
Souza, P. F. (2020). The forgotten 2S albumin proteins: Importance, structure, and biotechnological application in agriculture and human health. Int. J. Biol. Macromol. 4638-4649
Stills, H. F. (2012). Polyclonal antibody production. The Laboratory Rabbit, Guinea Pig, Hamster, and Other Rodents (pp. 259-274). Academic Press.
Stils Jr, H. F. (2005). Adjuvants and antibody production: dispelling the myths associated with Freund's complete and other adjuvants. ILAR journal, 46(3), 280-293.
Suprun, M., Sicherer, S. H., Wood, R. A., Jones, S. M., Leung, D. Y., Henning, A. K., ... & Suárez-Fariñas, M. (2020). Early epitope-specific IgE antibodies are predictive of childhood peanut allergy. J. Allergy Clin. Immunol. 146: 1080-1088.
Tian, Y., Liu, C., Zhang, K., Tao, S., & Xue, W. (2020). Glycosylation between recombinant peanut protein Ara h 1 and glucosamine could decrease the allergenicity due to the protein aggregation. LWT. 109374.
Tscheppe, A., Palmberger, D., van Rijt, L., Kalic, T., Mayr, V., Palladino, C., ... & Breiteneder, H. (2020). Development of a novel Ara h 2 hypoallergen with no IgE binding or anaphylactogenic activity. Journal of Allergy and Clinical Immunology, 145(1), 229-238.
Turasan, H., & Kokini, J. (2021). Novel Nondestructive Biosensors for the Food Industry. Annual Review of Food Science and Technology, 12, 539-566.
Turner, A. P. (2013). Biosensors: sense and sensibility. Chemical Society Reviews, 42(8), 3184-3196.
Walker, M. T., Green, J. E., Ferrie, R. P., Queener, A. M., Kaplan, M. H., & Cook-Mills, J. M. (2018). Mechanism for initiation of food allergy: dependence on skin barrier mutations and environmental allergen costimulation. J. Allergy Clin. Immunol. 141: 1711-1725.
Wang, W., Wang, E. Q., & Balthasar, J. P. (2008). Monoclonal antibody pharmacokinetics and pharmacodynamics. Clinical Pharmacology & Therapeutics, 84(5), 548-558.
Wingfield, P. T. (Ed.). (2016). Protein precipitation using ammonium sulfate. Current protocols in protein science, 84(1), A-3F.
Wu, Z., Yan, F., Wei, X., Li, X., Tong, P., Yang, A., ... & Chen, H. (2015). Purification and recombinant expression of major peanut allergen Ara h 1. Preparative Biochemistry and Biotechnology, 45(5), 438-446.
Wu, Z., Zhang, Y., Zhan, S., Lian, J., Zhao, R., Li, K., ... & Chen, H. (2017). Development of immunoaffinity chromatographic method for Ara h 2 isolation. Protein expression and purification, 131, 85-90.
Yan, Y.-S., Lin, X.-D., Zhang, Y.-S., Wang, L., Wu, K., & Huang, S.-Z. (2005). Isolation of peanut genes encoding arachins and conglutins by expressed sequence tags. Plant Sci. 169: 439–445.
Xia, X. (2007). Protein isoelectric point. Bioinformatics and the Cell: Modern Computational Approaches in Genomics, Proteomics and Transcriptomics, 207-219.
Xing, W., He, L., Yang, H., Sun, C., Li, D., Yang, X., ... & Deng, A. (2009). Development of a sensitive and group‐specific polyclonal antibody‐based enzyme‐linked immunosorbent assay (ELISA) for detection of malachite green and leucomalachite green in water and fish samples. Journal of the Science of Food and Agriculture, 89(13), 2165-2173.
Zhang, W., Zhu, Q., Zhang, T., Cai, Q., & Chen, Q. (2016). Thermal processing effects on peanut allergen Ara h 2 allergenicity in mice and its antigenic epitope structure. Food chemistry, 212, 657-662.
Zhao, L.,Zhao, L., Zhang, B., Robotham, J. M., Roux, K. H., & Tang, H. (2017). Identification of a common Ara h 3 epitope recognized by both the capture and the detection monoclonal antibodies in an ELISA detection kit. PloS one. 12: e0182935.