Establishment and performance evaluation of nanobody-based cat dander specific IgE antibody hypersensitivity assay
10.3969/j.issn.1000-484X.2024.04.025
- VernacularTitle:基于纳米抗体的猫皮屑特异性IgE抗体超敏检测方法的建立和性能评估
- Author:
Yanyan SUN
1
;
Desheng QI
;
Furao SHEN
;
Caiyue YANG
;
Xiangmei SUN
;
Guoning CHEN
;
Jianwei WU
Author Information
1. 济南德亨医学科技有限公司,济南 250031
- Keywords:
Nanobody;
Allergy;
Cat dander;
Specific IgE antibody;
Chemiluminescence
- From:
Chinese Journal of Immunology
2024;40(4):818-823
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To prepare anti-human IgE nanobody by phage display technology,and to establish a method for hyper-sensitivity detection of cat dander specific IgE antibody.Methods:Allergen bio-information of cat was searched in WHO/IUIS Allergen Database.After synthesizing sequence,recombinant cat dander allergenic protein Fel d 1 was expressed and purified in prokaryotic ex-pression system.Human IgE was used to immunize Bactrian camel and RNA were extracted from lymphocyte to construct phage dis-play library.Library capacity,diversity and insertion rate were analyzed,anti-human IgE nanobody were obtained by screening and identification.A magnetic particle chemical method for cat dander specific IgE antibody detection was established using recombinant allergen-coupled magnetic particles and acridine ester-labeled nanobodies.Results:Capacity of phage display library was 1.88×108 cfu/ml,insertion rate was 93.6%,and purity of nanobody was>95%.Linear range of the method based on nanobody was 0.1～100 U/ml,who was consistent with ImmunoCAP detection system by clinical data.Conclusion:Nanobody-based cat dander specific IgE antibody hypersensitivity assay is successfully prepared,providing a technical basis for auxiliary diagnosis of cat allergic diseases.
