Screen‐printed biosensor for allergens

Allergen levels in indoor environments, leading to many diseases, eg asthma, rhinitis and conjunctivitis, affect a large and increasing fraction of the population. A quite effective and inexpensive method of a rough but very rapid overall assessment of total allergen level in the environment has been developed. The method involved estimation of protein in allergen extracts by screen‐printed electrodes using two different techniques. The biosensor comprised a rhodinised carbon working electrode, a silver/silver chloride reference electrode and a carbon counter electrode. In the first method the enzyme protease reacted with allergen protein to release amino acid, which produced hydrogen peroxide in the presence of amino acid oxidase. This was detected amperometrically. The second method used potassium bromide as electrolyte and the electrode was subjected to dual potential. Bromine, released due to electrolysis at higher potential, was consumed by the allergen protein at lower potential. In the first method, a unique technique was used to microencapsulate the enzyme protease and immobilise it on the surface of the electrode by in‐situ polymerisation to avoid contact with the amino acid oxidase. A total of seven allergens were tested and the results gave a good correlation with the standard protein measurement method. Environmental specimens from indoors, schools and workplaces can be evaluated for the aeroallergens produced by dust mites, animal hairs, cockroach debris, pollens, etc as a means of determining the exposure risk. Copyright © 2005 Society of Chemical Industry     

食物致敏原表位定位技术的研究进展

食物致敏原是引起食物过敏的元凶，多为蛋白质。抗原表位是在抗原分子中与抗体反应或被抗原受体识别，并引发机体免疫应答的特殊化学基团。从表位水平认识食物致敏原，能揭示食物过敏的物质基础，为解决食物过敏问题提供精准的靶标。本文基于表位结构和定位方法的不同，介绍了食物致敏原表位定位技术的发展，并进一步展望了致敏原表位信息对于改进食物过敏鉴定技术和低致敏食物加工方法的应用前景。     

Isolation of recombinant antibody fragments (scFv) by phage display technology for detection of almond allergens in food products

Tree nut allergies are considered an important health issue in developed countries. To comply with the regulations on food labeling, reliable allergen detection methods are required. In this work we isolated almond-specific recombinant antibody fragments (scFv) from a commercial phage display library bypassing the use of live animals, hence being consistent with the latest policies on animal welfare. To this end an iterative selection procedure employing the Tomlinson I phage display library and a crude almond protein extract was carried out. Two different almond-specific scFv (named PD1F6 and PD2C9) were isolated after two rounds of biopanning, and an indirect phage ELISA was implemented to detect the presence of almond protein in foodstuffs. The isolated scFvs demonstrated to be highly specific and allowed detection of 40 ng mL⁻¹ and 100 ng mL⁻¹ of raw and roasted almond protein, respectively. The practical detection limit of the assay in almond spiked food products was 0.1 mg g⁻¹ (110–120 ppm). The developed indirect phage ELISA was validated by analysis of 92 commercial food products, showing good correlation with the results obtained by a previously developed real-time PCR method for the detection of almond in foodstuffs. The selected phage clones can be affinity maturated to improve their sensitivity and genetically engineered to be employed in different assay formats.     

食物过敏原检测与调控研究进展

食品过敏已成为一个重要的食品质量和安全问题,对食品加工行业构成挑战,并影响消费者的健康。一方面,从食品加工行业的角度来看,食品原料成分、外源添加剂和加工形式的多样性使得现代食品加工中过敏原的存在更加复杂。此外,由于缺乏过敏原识别和有效的检测与评价系统,导致目前食品过敏原筛选与检测、跟踪与预测、干预与控制的理论和技术存在严重不足;另一方面,从公共卫生的角度来看,满足消费者对包括食物过敏原在内的不同类型原料来源的知情权,提高政府的公信力和人民的满意度也成为当务之急;此外,随着人们接触的食物种类越来越多,食物过敏的发生概率也越来越高,日趋复杂化、广域化和严重化的食物过敏所带来的食品安全健康问题已很难避免。鉴于此,针对大健康背景下日益严重的食品过敏安全问题,本综述介绍了食物过敏原的检测方法,总结了食物过敏原消减与控制技术,阐述了低致敏性食品以及抗过敏活性物质,综述了目前食物过敏原口服免疫治疗进展,旨在为预防和控制食物过敏,保障食物过敏患者的身体健康提供依据。     

β-Conglycinin Reduces the Tight Junction Occludin and ZO-1 Expression in IPEC-J2

Soybean allergy presents a health threat to humans and animals. The mechanism by which food/feed allergen β-conglycinin injures the intestinal barrier has not been well understood. In this study, the changes of epithelial permeability, integrity, metabolic activity, the tight junction (TJ) distribution and expression induced by β-conglycinin were evaluated using IPEC-J2 model. The results showed a significant decrease of trans-epithelial electrical resistance (TEER) (p < 0.001) and metabolic activity (p < 0.001) and a remarkable increase of alkaline phosphatase (AP) activity (p < 0.001) in a dose-dependent manner. The expression levels of tight junction occludin and ZO-1 were decreased (p < 0.05). The reduced fluorescence of targets and change of cellular morphology were recorded. The tight junction occludin and ZO-1 mRNA expression linearly declined with increasing β-conglycinin (p < 0.001).     

Irritants and allergens at school in relation to furnishings and cleaning

In order to study the influence of furnishings and cleaning on the indoor air quality at school, 181 randomly chosen classrooms were investigated. The amounts of open shelves, textiles and other fittings were noted, data were gathered on cleaning routines, and a number of pollutants were measured in the classrooms. In classrooms with more fabrics there was more settled dust and the concentration of formaldehyde was higher. Classrooms with more open shelves had more formaldehyde, and more pet allergens in settled dust, and classrooms with a white board, instead of a chalk board, were less dusty. Classrooms mainly cleaned through wet mopping had more airborne viable bacteria but less settled dust than classrooms mainly cleaned by dry methods. In rooms where the desks and curtains were more often cleaned, the concentrations of cat and dog allergen in settled dust were lower. It is concluded that furnishings and textiles in the classroom act as significant reservoirs of irritants and allergens and have an impact on the indoor air quality at school.     

牛奶过敏原制备方法的研究和免疫活性鉴别

目的探索牛奶主要过敏原的制备工艺。方法采用等电点沉淀、凝胶层析和分子筛等技术纯化牛奶中主要过敏原组分;采用SDS-PAGE鉴定蛋白纯度,采用双抗原夹心-ELISA鉴定免疫活性。结果成功获得牛奶中的四种主要过敏原组分:酪蛋白、β乳球蛋白、α乳白蛋白和分子量较高的P1组分,并经过免疫实验证实这四种组分均能与过敏血清产生反应,而其中β乳球蛋白和P1组分反应性较高。结论本研究探索出一种简单、实用的牛奶主要过敏原制备工艺,并证实牛奶中的β乳球蛋白和高分子量蛋白质为主要过敏原。