Rapid Detection of Fish Major Allergen Parvalbumin by Surface Plasmon Resonance Biosensor
Rapid Detection of Fish Major Allergen Parvalbumin by Surface Plasmon Resonance Biosensor
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DOI：
10.1111/j.1750-3841.2004.tb18013.x
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发表时间：
2004-10
影响因子：
3.9
通讯作者：
Ying Lu;T. Ohshima;H. Ushio
中科院分区：
文献类型：
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作者：
Ying Lu;T. Ohshima;H. Ushio
海鲜过敏是成人食物过敏的常见和主要原因。近年来，随着海鲜消费量的增加，海鲜过敏已成为一个严重的问题。为了建立一种基于抗原-抗体相互作用亲和力的快速过敏原检测方法，采用表面等离子体共振（SPR）生物传感器对鱼类主要过敏原小清蛋白进行动力学分析。将鼠抗小清蛋白单克隆抗体（MAb）EGB固定在羧甲基葡聚糖（CMD）表面。通过注射不同浓度的纯化鲤鱼小白蛋白（GPa），测定了单克隆抗体EGB-CPa模型系统的标准曲线和亲和力常数（Kn和kag）。此外，动力学数据也得到了从海鲜产品的提取物的系列稀释液的注射：沙丁鱼鱼饼（tsumire）和干鲣鱼金枪鱼（katsuonut）。沙丁鱼tsumire和katsuonst分别含有0.11 mg/kg和0.39 mg/kg的parvalbumins，其中亲和常数Ks和kxes几乎是相似的parvalbomins从不同的来源。在SPR系统中，根据过敏原-MAb结合相互作用，过敏原仅可检测5 min。因此，通过使用SPR生物传感器，基于过敏原特异性单克隆抗体的动力学分析将是用于过敏原检测和定量的快速和强有力的工具。
Seafood allergy is a common and major cause of food allergy in aduits. In recent years, seafood allergy has become a serious problem with the increase of seafood consumption. To develop a rapid allergen detection method based on the affinity of antigen-antibody interaction, fish major allergen, parvalbumin, was used for kinetic analysis by a surface plasmon resonance (SPR) biosensor. Anti-parvalbumin murine monoclonal antibody (MAb) EGB was immobilized onto s carboxymethyl dextran (CMD) surface. By the injection of various concentrations of purified carp parvalibumin (GPa), a standard curve and the affinity consiants (K n and k ag ) for the MAb EGB-CPa model system were determined. In addition, kinetic data were also obtained by the injection of serial dilutions of extracts from seafood products: sardine fish cake (tsumire) and dried skipjack tuna (katsuonut). Sardine tsumire and katsuonst contained 0.11 mg/kg and 0.39 mg/kg parvalbumins, respectively, where affinity constants K s and k xes were almost similar among parvalbomins from different sources. In the SPR system, the allergen can be detected only for 5 min according to the allergen-MAb binding interaction. Consequently, by the use of a SPR biosensor, kinetic analysis based an the allergen specific MAb would be a rapid and powerful tool for allergen detection and quantification.