通过免疫亲和色谱法分离和鉴定针对鸡蛋清溶菌酶的蛋黄免疫球蛋白（IgY）

Isolation and characterization of immunoglobulin in yolk (IgY) specific against hen egg white lysozyme by immunoaffinity chromatography.

作者信息

Chen Chao-Cheng, Tu Yann-Ying, Chen Tzy-Li, Chang Hung-Min

机构信息

Department of Food Health, Chia-Nan University of Pharmacy and Science, Tainan 717, Taiwan.

出版信息

J Agric Food Chem. 2002 Sep 11;50(19):5424-8. doi: 10.1021/jf011567h.

DOI:10.1021/jf011567h

PMID:12207486

Abstract

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.

摘要

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。