Screening of Nanobody Specific for Peanut Major Allergen Ara h 3 by Phage Display

被引：29

作者：

Chen, Feng ^{[1
]}

Ma, Huan ^{[1
]}

Li, Yuelong ^{[1
]}

Wang, Heqiao ^{[1
]}

Samad, Abdus ^{[1
]}

Zhou, Jiahui ^{[1
]}

Zhu, Lixia ^{[1
]}

Zhang, Yuzhu ^{[2
]}

He, Jie ^{[3
]}

Fan, Xiaojiao ^{[1
]}

Jin, Tengchuan ^{[1
]}

机构：

[1] Univ Sci & Technol China, Div Life Sci & Med, CAS Key Lab Innate Immun & Chron Dis, Hefei Natl Lab Phys Sci Microscale, Hefei 230027, Anhui, Peoples R China

[2] USDA ARS, Hlth Proc Foods Res Unit, Western Reg Res Ctr, 800 Buchanan St, Albany, CA 94710 USA

[3] USTC, Affiliated Hosp 1, Div Life Sci & Med, Dept Pathol, Hefei 230001, Anhui, Peoples R China

来源：

JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY | 2019年 / 67卷 / 40期

关键词：

nanobody; Ara h 3; peanut allergen; food safety; allergen detection; FOOD ALLERGY; IGE; ANTIBODIES; DIAGNOSIS; CHILDREN;

D O I：

10.1021/acs.jafc.9b02388

中图分类号：

S [农业科学];

学科分类号：

09 ;

摘要：

Peanut allergy is a major health problem worldwide. Detection of food allergens is a critical aspect of food safety. The VHH domain of single chain antibody from camelids, also known as nanobody (Nb), showed its advantages in the development of biosensors because of its high stability, small molecular size, and ease of production. However, no nanobody specific to peanut allergens has been developed. In this study, we constructed a library with random triplets (NNK) in its CDR regions of a camel nanobody backbone. We screened the library with peanut allergy Ara h 3 and obtained several candidate nanobodies. One of the promising nanobodies, Nb16 was further biochemical characterization by gel filtration, isothermal titration calorimetry (ITC), cocrystallization, and Western blot in terms of its interaction with Ara h 3. Nb16 specifically binds to peanut major allergen Ara h 3 with a dissociation constant of 400 nM. Furthermore, we obtained the Ara h 3-Nb16 complex crystals. Structure analysis shows the packing mode is completely different between the Ara h 3-Nb16 complex crystal and the native Ara h 3 crystal. Structural determination of Ara h 3-Nb16 will provide the necessary information to understand the allergenicity of this important peanut allergen. The nanobody Nb16 may have application in the development of biosensors for peanut allergen detection.

引用

页码：11219 / 11229

页数：11

共 37 条

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