Frontiers in Food Allergy and Allergen Risk Assessment and Management. 19 April 2018, Madrid

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Frontiers in Food Allergy and Allergen Risk Assessment and Management 19 April 2018, Madrid

Food allergy is becoming one of the serious problems of China's food safety and public health emergency.

7 Number of published papers 6 5 4 3 2 1 0 2000 2005 2009 2010 2011 2012 2013 2015 Published year

The prevalence of food allergy seems to have increased in China. Using oral food challenges for diagnosis, two crosssectional studies of infants from the same clinic in China showed that food allergy prevalence increased significantly from 3.5% in 1999 to 7.7% in 2009. Source: Hu Y, et al. Comparison of food allergy prevalence among Chinese infants in Chongqing, 2009 versus 1999. Pediatr Int 2010;52:820-4.

Labels are the only way for consumers to know about prepackaged food ingredients.

Big-8, as recommended by Codex, includes eggs, fish, milk, peanuts, shellfish, soybeans, tree nuts and wheat. These foods account for about 90% of all food allergies. Labelling of the Big-8 is mandatory according to EU, Australian/New Zealand, Canadian and the USA regulations.

GB 7718-2011 National Food Safety Standard -- General Standard for the Labelling of Prepackaged Foods

GB/T 23779-2009 Allergenic Ingredients in Prepackaged Foods

Allergenic Food Labelling is volunteering in China. Big-8 in China follow Codex recommendations, lacking of supporting from epidemiological data on food allergy in Chinese population.

Focused on the key issues of food allergy in the comparative study of domestic and foreign technology in food safety. To set up a food allergy work group. To carry out food allergy epidemiological studies. To establish food labelling system for food allergen that is more in line with China's national conditions.

The overall arrangements for the project on food allergy risk assessment in China during the 13 th Five- Year Plan period was preliminarily determined.

Types of allergenic foods Characteristics of allergens Validation and assessment Risk control FA diagnostic criteria Diagnostic reagents Validation Food allergen validation Chinese population Major allergenic foods in China Major allergens identification Epitope structure and function Detect reagents Standard candidates Epidemiological studies Allergenicity assessment Series of technical standards on Food allergens Serum bank Risk management technology system on food allergy labelling Feedback

1. Multi-center epidemiologic studies on food allergy in Chinese population 2. Epitope structure and allergenic property of food allergens 3. Multi-level technology system for allergenicity assessment on food allergens 4. The validation technology of the testing food allergens and related product development 5. Research and development on food allergy clinical diagnostic key reagents 6. Research on food allergy risk management measures

Nationwide epidemiological studies of food allergies are conducted, including cross-sectional study (31 provinces, 10 provinces) prospective birth cohort study (4 provinces) nested case-control study (4 provinces)

To determine the prevalence of food allergy. To identify major allergenic foods in the population, providing support for updating the allergenic food labelling list in China. To identify the risk factors for the development of food allergies. 15

Questionnaires (31 provinces) Skin prick test Measurement of food specific IgE Food allergy diagnostic criteria needs to be updated, with the inclusion of the gold standard diagnostic test, i.e., double-blind, placebocontrolled food challenge. 16

Epitope structure and allergenic property of food allergens Aims Mapping of T cell epitopes and B cell epitopes Mapping of the key amino acid Elucidating immunological significance of allergens and epitopes in food allergy in Chinese population Exploring the common and similar epitopes for immune cross-reaction to reveal mechanisms of allergic cross-reaction Elucidating the effect of changes in epitope structure on allergenicity, providing structural information for allergenicity evaluation of processed foods.

The relationship between structure and allergenicity of main allergens that stimulate Chinese food allergy Study on the mapping, structure and immunological characteristics of food allergen epitopes The role of food allergen epitopes in immune cross-reaction

Food allergens Digestion assessment Digestion model Digestion resistant peptide Processing on amino acid modification Processing Critical amino acid modification In Vitro Assessment Dendritic cell T cell B cell Mast cell Animal model In Vivo Assessment Relationship Analysis Multi-level technology system for allergenicity assessment on food allergens

The validation technology of the testing food allergens and related product development Food allergens identification database Peptide profiles acquisition using high resolution mass spectrometry Comparison of theoretical and actual peptide sequence Establishment of food allergens identification database Food allergens quantification methods Proteotypic peptide selection of specific food allergens Optimization of pretreatment for food allergens in processed food Establishment of food allergens quantification method Food allergens identification and quantitative kits Development of food allergens determination kits Performance validation of food allergens determination kits Comparison of our products with merchant ELISA kits

Food allergens identification database Peptide profiles acquisition using high resolution mass spectrometry UHPLC-Q- Orbitrap UPLC-Q-TOF Comparison of theoretical and actual peptide sequence Establishment of food allergens identification database

Food allergens quantification methods Workflow of Targeted Proteomics DTT IAA Trypsin Orbitrap RT: 0.00-10.00 Relative Abundance 100 90 80 70 60 50 40 30 20 10 0 0.87 2.86 1.15 1.23 2.89 2.84 2.93 3.37 2.62 2.60 3.52 3.69 4.19 1.47 7.13 7.11 7.17 4.25 4.71 8.48 4.81 8.50 4.87 8.51 5.05 8.55 8.65 9.04 0 2 4 6 8 10 Time (min) NL: 1.38E9 m/z= 150.00000-2000.00000 F: FTMS + p ESI Full ms MS 20160320-BSAddMS-1 Peptide selection

Food allergens quantification methods Workflow of Targeted Proteomics MRM Triple Quadrupole Intensity [counts] (10^6) Human-d10-ddMS.raw #2532 RT: 5.1780 min FTMS, 483.2580@hcd30.00, z=+2, Mono m/z=483.25790 Da, MH+=965.50853 Da, Match Tol.=0.02 Da 1.4 y₅+ 1.2 607.35577 1.0 72.08154 y₄+ 508.28784 0.8 b₃+ 260.08771 0.6 y₁+ b₃+-h₂o 159.11273 175.11887 242.07689 y₃+ b₂+ 608.35901 345.22430 0.4 173.05571 y₁+-nh₃ b₄+-h₂o 509.28998 158.09206 341.14551 y₄+-nh₃ y₆+ y₇+ 0.2 491.25995 y₅+-nh₃ 706.42188 793.45612 y₂ b₄ 590.32507 707.42432 510.28903 0.0 100 200 300 400 500 600 700 800 900 1000 m/z (i) Specificity (ii) Digestibility (iii) Mass characteristics Peptide selection

Food allergens identification and quantitative kits Development of food allergens determination kits Based on methods and results of above researches Performance validation of food allergens determination kits Including method validation, precision, sensitivity, batch stability, etc. Comparison of our products with merchant ELISA kits

To decrease medical expense, the most common food allergen extracts for diagnosing food allergy are needed to be developed and to be approved by the China Food and Drug Administration.

Research on risk management measures for food allergy Risk Assessment To establish a food allergy risk assessment model. Risk Management To formulate or revise a series of technical standard system.

The establishment of the food allergy risk assessment model Risk identification and risk monitoring A To determine the key factors in the model B To establish the food allergy risk assessment mathematical model E Validation and correction Determine the key parameter values C D Analysis and evaluation 27

The formulation and revision of the technical standard system on the risk management of food allergy. To formulate/reformulate standard system. To make suggestions and to formulate measures. To analyze the present situation in China. To summarize laws and regulations in other countries

Dr. Hongbing Chen, State Key Laboratory of Food Science and Technology, Nanchang University, Nanchang, China Dr. Zhenxing Li, College of Food Science and Engineering, Ocean University of China, Qingdao, China Dr. Yiping Ren, Yangtze Delta Region Institute of Tsinghua University, China National Center for Food Safety Risk Assessment, Jiaxing, China Dr. Jinlu Sun, Department of Allergy, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences, Beijing, China Dr. Huilian Che, College of Food Science and Nutritional Engineering, China Agricultural University, Beijing, China Dr. Yongning Wu, Key Laboratory of Food Safety Risk Assessment of National Health Commission, China National Center for Food Safety Risk Assessment, Beijing, China