公开(公告)号：US06677433B2

公开(公告)日：2004-01-13

申请号：US09779375

申请日：2001-02-07

申请人： Bob B. Buchanan ,  Susumu Morigasaki ,  Gregorio del Val ,  Oscar L. Frick

发明人： Bob B. Buchanan ,  Susumu Morigasaki ,  Gregorio del Val ,  Oscar L. Frick

IPC分类号： C07K1400

CPC分类号： A21D2/26 ,  A21D2/264 ,  A21D2/265 ,  A21D2/266 ,  A21D2/36 ,  A21D8/04 ,  A21D13/064 ,  A23C9/1209 ,  A23C9/1213 ,  A23C9/152 ,  A23J1/12 ,  A23J1/14 ,  A23J3/08 ,  A23J3/18 ,  A23K20/147 ,  A23L5/27 ,  A23L7/109 ,  A23L11/34 ,  A23L13/48 ,  A23L33/17 ,  A61K38/00 ,  A61K39/35 ,  A61K2035/122 ,  C07K1/1133 ,  C07K14/415 ,  C07K14/43504 ,  C07K14/46 ,  C07K14/76 ,  C07K14/811 ,  C12N9/0036 ,  C12N9/20 ,  C12P21/02 ,  C12Q1/26 ,  C12Q1/34 ,  C12Y108/01008 ,  C12Y108/01009

摘要： Disulfide proteins showed mitigated allergenicity and increased digestibility by pepsin following reduction by thioredoxin. The sulfhydryl groups newly formed on reduction by thioredoxin (at 4° C.) or dithiothreitol (DTT) (at 55° C.) were blocked with a physiological disulfide, such as cystamine or oxidized glutathione (GSSG) to obtain stable forms of the disarmed allergen. When derivatized with cystamine, BLG was separated from its oxidized and reduced forms on non-reducing SDS-PAGE and appeared to lack sulfhydryl groups. Although less effective GSSG, gave similar results. Allergenicity of the two derivatives was compared with that of the oxidized, reduced and reoxidized forms of BLG by skin testing dogs from a colony sensitized to cow's milk. Both the cystamine and GSSG derivatized BLG showed decreased allergenicity and increased sensitivity to pepsin as compared to controls. The reoxidized form resembled the derivatives in having lower allergenicity. The thioredoxin- and DTT-reduced forms showed hypoallergenic, hyperdigestible properties, most effectively when the reduced proteins were heated at 55° C. Whole milk subjected to these procedures showed results similar to those obtained with pure BLG. Other proteins are similarly stabilized. Stable forms of such disarmed, hypoallergenic and hyperdigestible disulfide protein allergens or just hypoallergenic or just hyperdigestible protein allergens are useful in foods as well as clinical preparations.

摘要翻译： 二硫化物蛋白质在硫氧还蛋白还原后显示减轻的过敏原性和胃蛋白酶的消化率增加。 通过硫氧还蛋白（4℃）还原二硫苏糖醇（DTT）（在55℃）下新形成的巯基用生理二硫化物如胱氨酸或氧化型谷胱甘肽（GSSG）封闭，得到稳定形式的 解除武装的过敏原 当用胱胺衍生化时，BLG在非还原性SDS-PAGE上从其氧化和还原形式分离出来，并且似乎缺少巯基。 虽然GSSG效果较差，但得到了类似的结果。 将两种衍生物的过敏原与来自对牛奶敏感的菌落进行皮肤测试的狗的氧化，还原和再氧化形式的BLG进行比较。 与对照相比，胱氨酸和GSSG衍生的BLG都显示出降低的过敏原性和对胃蛋白酶的敏感性增加。 再氧化形式类似于具有较低过敏原性的衍生物。 当减少的蛋白质在55℃下加热时，硫氧还蛋白和DTT减少的形式显示出低变应原性，超消化性质，最有效的是，经过这些程序的全脂乳显示出与用纯BLG获得的结果相似的结果。 其他蛋白质同样稳定。 这种解除武装，低变应原和超可消化的二硫化物蛋白质过敏原或刚刚低变应原或仅仅是超可消化的蛋白质过敏原的稳定形式可用于食物以及临床制剂。