Babij Konrad, Bajzert Joanna, Dąbrowska Anna, Szołtysik Marek, Zambrowicz Aleksandra, Lubec Gert, Stefaniak Tadeusz, Willak-Janc Ewa, Chrzanowska Józefa
Department of Animal Products Technology and Quality Management, Wrocław University of Environmental and Life Sciences, ul. Chełmońskiego 37/41, 51-630, Wrocław, Poland.
Department of Immunology, Pathophysiology and Veterinary Preventive Medicine, Wrocław University of Environmental and Life Sciences, Wrocław, Poland.
Amino Acids. 2015 Nov;47(11):2335-43. doi: 10.1007/s00726-015-2013-2. Epub 2015 Jun 3.
In the present study the effect of hydrolysis with non-commercial Cucurbita ficifolia serine protease on a reduction of the IgE and IgG binding capacity of whey protein concentrate and αs-casein was investigated. The intensity of the protein degradation was analyzed by the degree of hydrolysis, the free amino groups content and RP-HPLC. The ability to bind the antibodies by native proteins and their hydrolysates was determined using a competitive ELISA test. Deep hydrolysis contributed to a significant reduction of immunoreactive epitopes present in WPC. In the case of IgE and IgG present in the serum pool of children with CMA, the lowest binding capacity was detected in the 24 h WPC hydrolysate, where the inhibition of the reaction with native WPC was ≤23 and ≤60 %, respectively. The analysis of the IgG reactivity in the antiserum of the immunized goat showed that the lowest antibody binding capacity was exhibited also by 24 h WPC hydrolysate at a concentration of 1000 μg/ml where the inhibition of the reaction with nWPC was ≤47 %. One-hour hydrolysis of α-casein was sufficient to significant reduction of the protein antigenicity, while the longer time (5 h) of hydrolysis probably lead to the appearance of new epitopes reactive with polyclonal.
在本研究中，研究了用非商业性的黑籽南瓜丝氨酸蛋白酶水解对降低乳清蛋白浓缩物和αs-酪蛋白的IgE和IgG结合能力的影响。通过水解度、游离氨基含量和反相高效液相色谱法分析蛋白质降解的强度。使用竞争性酶联免疫吸附测定法测定天然蛋白质及其水解产物结合抗体的能力。深度水解导致乳清蛋白浓缩物中存在的免疫反应性表位显著减少。对于患有牛奶蛋白过敏症儿童血清库中的IgE和IgG，在24小时乳清蛋白浓缩物水解产物中检测到最低的结合能力，其中与天然乳清蛋白浓缩物反应的抑制率分别≤23%和≤60%。对免疫山羊抗血清中IgG反应性的分析表明，在浓度为1000μg/ml的24小时乳清蛋白浓缩物水解产物中也表现出最低的抗体结合能力，其中与天然乳清蛋白浓缩物反应的抑制率≤47%。α-酪蛋白1小时的水解足以显著降低蛋白质的抗原性，而较长时间（5小时）的水解可能导致出现与多克隆抗体反应的新表位。
