Chen Chao-Cheng, Tu Yann-Ying, Chen Tzy-Li, Chang Hung-Min
Department of Food Health, Chia-Nan University of Pharmacy and Science, Tainan 717, Taiwan.
J Agric Food Chem. 2002 Sep 11;50(19):5424-8. doi: 10.1021/jf011567h.
Six hens were intramuscularly (im) immunized once a week for 3 weeks using chicken egg white lysozyme (LS) as antigen. Antibody (immunoglobulin in yolk, IgY) ELISA values of 10(3)-fold diluted yolk were almost as high as 1.879 in the sixth week and maintained a value of 0.756 in the eighth week after the initial immunization treatment. The purification efficiency (specific activity of purified IgY against LS/specific activity of antibody in yolk against LS) of IgY specific against LS isolated by laboratory-prepared LS-bound (IgY-) Sepharose 4 Fast Flow immunoaffinity column was approximately 3380. By applying various amounts (0-22 mg) of the thusly obtained IgY specific against LS to the immunoaffinity column, the binding capacity (q(m)) and dissociation constant (K(d), M(-1)) of such immunoaffinity gel for IgY against LS were found to be 0.68 mg of IgY/mL of wet gel (0.54 mg of IgY/mg of LS) and 7.13 x 10(-6) M, respectively, as determined by Langmuir-type adsorption isotherms.
以鸡卵清溶菌酶（LS）作为抗原，对6只母鸡进行肌肉注射免疫，每周1次，共3周。初次免疫处理后，第6周时10³倍稀释卵黄的抗体（卵黄中的免疫球蛋白，IgY）ELISA值几乎高达1.879，第8周时维持在0.756。通过实验室制备的LS结合（IgY-）琼脂糖4快速流动免疫亲和柱分离得到的抗LS特异性IgY的纯化效率（纯化IgY对LS的比活性/卵黄中抗体对LS的比活性）约为3380。通过将不同量（0 - 22 mg）的上述抗LS特异性IgY应用于免疫亲和柱，根据朗缪尔型吸附等温线测定，该免疫亲和凝胶对抗LS的IgY的结合容量（q(m)）和解离常数（K(d)，M⁻¹）分别为0.68 mg IgY/mL湿凝胶（0.54 mg IgY/mg LS）和7.13×10⁻⁶ M。
