Construction of the fundamental data for the clinical test of the allergy patients' substituted foods

卵清蛋白（OVA）和卵黏液样蛋白（OM）是蛋清的主要过敏原，常引起过敏性儿童的过敏反应。日本鹌鹑（JQ）蛋OVA^<1)>或OM^<2)>与母鸡(H)蛋OVA^<3)>或OM^<4)>序列同源性分别为88.9%和75.8%。抑制ELISA法测定JQ-OVA和JQ-OM对兔抗H-OVA血清和抗H-OM血清的结合活性分别为6.8%和0.04%。这些结果表明，当几个氨基酸残基相互取代时，抗原性显著降低。本研究旨在通过免疫印迹和微序列分析方法表征JQ-OVA的抗原决定因素。为此，采用硫酸铵分离和凝胶过滤层析，从JQ蛋白中分离出JQ- ova。将纯化的JQ-OVA注射到母兔体内制备抗JQ-OVA血清。JQ-OVA用金黄色葡萄球菌V8蛋白酶、tpck -胰蛋白酶、tlck -糜凝胰蛋白酶和胃蛋白酶进行酶切，用三辛sds -聚丙烯酰胺凝胶电泳和二维凝胶电泳分离肽段。通过结合抗JQ-OVA兔IgG抗体检测其抗原性。推测JQ-OVA 17-28、72-84、1 20-124、132-149、160-175、206-211、219-226、233-242、244-253和298-308序列中存在部分抗原决定因子（或表位）。这些表位与H-OVA的表位不同。此外，婴儿食品（茶饭、蛋奶布丁、蛋奶汤）都是用JQ-egg和H-egg制作的。一项感官测试显示，这些用j - q -egg和H-egg烹制的婴儿食品没有区别。这些结果将为鸡蛋过敏提供更好的治疗方法。1)Nisbet等，1981欧元。[2]张晓明，张晓明，张晓明，等。生物化学学报，1997,26(3):335-345。[4]张建军，张建军，张建军，等。， 158, 515-537

Ovalbumin (OVA) and ovomucoid (OM) are major allergens of hen's egg white and often induce hypersensitivity in atopic children. The sequence homology of Japanese quail (JQ) egg OVA^<1)> or OM^<2)> and hen (H) egg OVA^<3)> or OM^<4)> is 88.9% or 75.8% respectively. But the binding activities of JQ-OVA to rabbit anti H-OVA serum and JQ-OM to anti H-OM serum, measured by the method of inhibition ELISA, were 6.8% and 0.04% respectively. These results indicate that the antigenicity decreases dramatically when a few amino acid residues replace each other. This study aimed to characterize the antigenic determinants of JQ-OVA by immunoblotting and microsequence analysis methods. For this purpose, JQ-OVA was isolated from the JQ egg white by the fractionation with ammonium sulfate and gel filtration chromatography. The rabbit anti JQ-OVA serum was prepared by injecting the purified JQ-OVA to a female rabbit. JQ-OVA was digested with S. aureus V8 protease, TPCK-trypsin, TLCK-chymotrypsin and pepsin, and the peptides were separated with tricine SDS-polyacrylamide gel electrophoresis and two-dimentional gel electrophoresis. The antigenicity was tested by binding an anti JQ-OVA rabbit IgG antibody with the peptides. It is proposed that a part of the antigenic determinant (or epitope) existed in the JQ-OVA sequences of 17-28, 72-84, 1 20-124, 132-149, 160-175, 206-211, 219-226, 233-242, 244-253 and 298-308. These epitopes differed from those of H-OVA. Furthermore, baby foods (chawan-musi, custard pudding egg bouro, and kakitama soup) were cooked with JQ-egg and H-egg. A sensory test revealed that there were no differences between these baby foods cooked with JQ-egg and H-egg. Those results will allow better treatment to be provided for the hen egg allergy.1)Nisbet, et al. 1981 Eur. J. Biochem, 115, 335-345, 2)Kato, et al. 1987 Biochemistry, 26, 193-2013)Mucha,et al. 1990 Nucleic Acids Res., 18, 5553. 4)Papamakos, et al. 1982 J. Ml. Biol., 158, 515-537