Kato Y, Oozawa E, Matsuda T
Department of Clinical Nutrition, Medical Professions, Kawasaki University of Medical Welfare, 288 Matsushima, Kurashiki, 701-0193, Japan.
J Agric Food Chem. 2001 Aug;49(8):3661-5. doi: 10.1021/jf0102766.
The antigenic and allergenic activities of ovomucoid (OM) remaining in soluble fractions of pasta-like model samples of wheat flour mixed with egg white were investigated by ELISA competitive inhibition and immunoblotting analyses using a rabbit anti-OM IgG and the serum IgE specific for OM in patients allergic to egg white. The mixture of egg white and wheat flour of soft, hard, and durum varieties was kneaded for 10-50 min and benched for 1 h at RT, and then small pieces of the dough were heated in boiling 1% NaCl solution for 15 min. Even before heating, only after the kneading for 30 min or more, but not after kneading for only 20 min, followed by the benching, the antigenic activity of OM which remained in the phosphate-buffered saline extract from the dough markedly decreased. Almost no antigenic activity of OM was detected in the extracts of heated samples. Furthermore, in the extracts of heated durum samples, only a trace of or almost no IgE-reactive OM was detected against the five patients' sera. These reductive effects of wheat on the OM antigenicity and allergenicity were more remarkable in the durum variety than in the others. No detectable proteins were extracted with 1% SDS from the heated samples, whereas OM was extracted with 1% SDS containing 10% 2-mercaptoethanol, suggesting heat-induced polymerization through intermolecular disulfide bonds among OM and wheat.
通过酶联免疫吸附试验(ELISA)竞争抑制和免疫印迹分析,使用兔抗卵类粘蛋白(OM)IgG以及对蛋清过敏患者血清中针对OM的特异性IgE,研究了与蛋清混合的小麦粉类意大利面模型样品可溶部分中残留的OM的抗原活性和致敏活性。将软质、硬质和硬质小麦品种的蛋清与小麦粉混合物揉捏10 - 50分钟,室温放置1小时,然后将小块面团在沸腾的1%氯化钠溶液中加热15分钟。即使在加热之前,仅在揉捏30分钟或更长时间后,而不是仅揉捏20分钟后再放置,面团磷酸盐缓冲盐水提取物中残留的OM抗原活性就会显著降低。在加热样品的提取物中几乎检测不到OM的抗原活性。此外,在加热的硬质小麦样品提取物中,针对五名患者血清,仅检测到微量或几乎没有IgE反应性OM。小麦对OM抗原性和致敏性的这些还原作用在硬质小麦品种中比在其他品种中更显著。加热样品用1%十二烷基硫酸钠(SDS)提取不到可检测的蛋白质,而OM用含10% 2 - 巯基乙醇的1% SDS提取,这表明加热通过OM与小麦之间的分子间二硫键诱导聚合。