Development of a monoclonal antibody-based sandwich ELISA for detection and quantification of the major cat allergen Fel d 1

文献类型： 外文期刊

第一作者： Bi, Zhuangli

作者： Bi, Zhuangli;Cai, Linying;Zhang, Zichen;Du, Qingqing;Zhu, Shiqiang;Zhang, Miao;Fan, Yiming;Tang, Jingjie;Hu, Mingxing;Tang, Aoxing;Liu, Guangqing;Zhu, Yingqi;Cai, Linying;Zhang, Zichen;Du, Qingqing;Tang, Jingjie;Hu, Mingxing;Wang, Guijun

作者机构：

关键词： Felis domestic allergen 1 (Fel d 1); Monoclonal antibodies (mAbs); Sandwich ELISA

期刊名称：INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES （ 影响因子：8.5； 五年影响因子：8.7 ）

ISSN： 0141-8130

年卷期： 2025 年 323 卷

页码：

收录情况： SCI

摘要： Felis domesticus allergen 1 (Fel d 1), the primary allergen produced by cats, is a key trigger of various allergic symptoms in humans, such as rhinitis, conjunctivitis, and asthma. In this study, a recombinant Fel d 1 protein composed of chain 2 and chain 1 was successfully produced in E. coli, based on a previously reported fusion construct that closely approximates the native allergen. Two antibodies, 2A7 and 6B1, were selected for further study due to their high specificity and complementary epitope recognition. A dual-antibody sandwich ELISA was established using 2A7 as the capture antibody and HRP-conjugated 6B1 as the detection antibody. The assay showed a detection range of 10-500 ng/mL with a sensitivity of 1 ng/mL. Application to feline saliva and fur samples revealed detectable Fel d 1 levels, and results correlated well with commercial kits. Epitope mapping identified distinct alpha-helical regions targeted by 2A7 and 6B1, and sequence analysis of their variable regions confirmed diverse V(D)J gene usage. This study provides a valuable antibody pair and a sensitive ELISA method for Fel d 1 quantification, with potential applications in allergen exposure assessment and diagnostic development. Scientific section: Immunology, Antibodies, and Diagnostics.

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