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针对屋尘螨蛋白质的蛋黄抗体的制备。

Production of egg yolk antibodies specific to house dust mite proteins.

作者信息

Lee Kyung Eun, Han Beom Ku, Han Jae Yong, Hong Jung Yeon, Kim Mi Na, Heo Won Il, Sohn Myung Hyun, Kim Kyung Won, Kim Kyu Earn

机构信息

Department of Pediatrics and Institute of Allergy, Brain Korea 21 Plus Project for Medical Science, Yonsei University College of Medicine, Seoul, Korea.

Optipharm Inc., Osong, Korea.

出版信息

Yonsei Med J. 2014 Jul;55(4):999-1004. doi: 10.3349/ymj.2014.55.4.999.

Abstract

PURPOSE

House dust mites (HDMs) are an important source of indoor allergens associated with asthma, rhinitis and atopic dermatitis. Chicken immunoglobulin (Ig) Y is known to be a good alternative to mice and rabbit antibody production. In this study, we produced IgYs specific to HDMs and investigated their IgE immunoreactivities.

MATERIALS AND METHODS

Total IgYs were isolated from the yolks of White Leghorn hens immunized with either Dermatophagoides pteronyssinus or D. farinae protein extract. Control antibodies were separated from the yolks of immunized hens with phosphate buffered saline. IgYs specific to HDMs were analyzed using enzyme-linked immunosorbent assay and Western blotting analysis.

RESULTS

The concentration of egg IgY specific to D. farinae in an immunized hen increased and the highest achieved was 661.3 ug/mg (per an egg) on day 47, compared with 760 ug/mg IgY specific to D. pteronyssinus on day 16. The D. pteronyssinus or D. farinae-specific IgY was detected by binding of each mite proteins, and their immunoreactivities were elevated dependent of the specific IgY concentration.

CONCLUSION

IgY specific to HDMs may be a promising antibody for immunological diagnosis as well as identification of possible resistance relating to HDM allergy.

摘要

目的

屋尘螨(HDMs)是与哮喘、鼻炎和特应性皮炎相关的室内过敏原的重要来源。鸡免疫球蛋白(Ig)Y是已知的替代小鼠和兔抗体生产的良好选择。在本研究中,我们制备了针对屋尘螨的IgY,并研究了它们的IgE免疫反应性。

材料与方法

从用粉尘螨或粗脚粉螨蛋白提取物免疫的白来航鸡的蛋黄中分离总IgY。对照抗体从用磷酸盐缓冲盐水免疫的母鸡的蛋黄中分离。使用酶联免疫吸附测定和蛋白质印迹分析来分析针对屋尘螨的IgY。

结果

免疫母鸡中针对粗脚粉螨的卵IgY浓度增加,在第47天达到最高值661.3μg/mg(每枚蛋),而针对粉尘螨的IgY在第16天为760μg/mg。通过每种螨蛋白的结合检测到粉尘螨或粗脚粉螨特异性IgY,并且它们的免疫反应性随着特异性IgY浓度的增加而升高。

结论

针对屋尘螨的IgY可能是用于免疫诊断以及鉴定与屋尘螨过敏相关的可能抗性的有前景的抗体。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3c36/4075406/5d8d26e4818d/ymj-55-999-g001.jpg

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