Generation and application of anti-ouabain IgY antibodies

被引:2
|
作者
Zhang, Ming Juan [1 ]
Yang, Jun [2 ]
Zhu, Can Zhan [1 ]
Duan, Zong Ming [1 ]
Niu, Xiao Lin [1 ]
机构
[1] Xi An Jiao Tong Univ, Affiliated Hosp 2, Sch Med, Dept Cardiol, Xian 710004, Peoples R China
[2] Xi An Jiao Tong Univ, Affiliated Hosp 2, Sch Med, Dept Pathol, Xian 710004, Peoples R China
基金
中国国家自然科学基金;
关键词
Adrenal; Immunohistochemistry; IgY (yolk immunoglobulin); Ouabain; Titer; BOVINE ADRENOCORTICAL-CELLS; FETAL ADRENAL-GLAND; ENDOGENOUS OUABAIN; HUMAN PLASMA; IDENTIFICATION; APOPTOSIS; INHIBITOR; COMPOUND; RECEPTOR; GROWTH;
D O I
10.1007/s11010-011-0974-3
中图分类号
Q2 [细胞生物学];
学科分类号
071009 ; 090102 ;
摘要
Ouabain is a bioactive hapten and is very difficult to be accurately quantified because of the lack of useful reagents. Furthermore, where ouabain is produced in the adrenal glands has not been identified. In this study, ouabain-BSA was generated for immunizing the laying hens to generate ouabain-specific IgY antibodies in chicken eggs. The anti-ouabain IgY antibodies were detected in eggs 1 week after the last immunization and their concentrations increased with time. The highest concentrations of anti-ouabain IgY antibodies reached at 1:10,240 for ELISA 5 weeks after immunization and maintained for 4 weeks in chicken eggs. Following PEG precipitation, an average of 8.5 mg of anti-ouabain IgY antibodies with a purity of 87.6% was achieved from a single egg. Further analysis revealed that the anti-ouabain IgY antibodies had little immunoreactivity to hydrocortisone, dexamethasone, cedilanid, and digoxin, indicating their high specificity, and the purified IgY antibodies effectively detected endogenous ouabain in the cytoplasm of cells predominately in the zona reticularis of rat and human adrenal glands, indicating their high immunoreactivity. Given that IgY has an unique structure and bioactive features, the generated anti-ouabain IgY antibodies may be used as a new reagent for accurately quantifying ouabain in biological studies.
引用
收藏
页码:241 / 247
页数:7
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