Skip to main content
ARS Home » Pacific West Area » Albany, California » Western Regional Research Center » Healthy Processed Foods Research » Research » Publications at this Location » Publication #368650
Research Project: Defining, Measuring, and Mitigating Attributes that Adversely Impact the Quality and Marketability of Foods

Location: Healthy Processed Foods Research

Title: Screening of nanobody specific for peanut major allergen Ara h 3 by phage display

Author
item CHEN, FENG - University Of Science And Technology Of China
item MA, HUAN - University Of Science And Technology Of China
item LI, YUELONG - University Of Science And Technology Of China
item WANG, HEQIAO - University Of Science And Technology Of China
item SAMAD, ABDUS - University Of Science And Technology Of China
item ZHOU, JIAHUI - University Of Science And Technology Of China
item ZHU, LIXIA - University Of Science And Technology Of China
item Zhang, Yuzhu
item HE, JIE - University Of Science And Technology Of China
item FAN, XIAOJIAO - University Of Science And Technology Of China
item JIN, TENGCHUAN - University Of Science And Technology Of China

Submitted to: Journal of Agricultural and Food Chemistry
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 8/13/2019
Publication Date: 8/13/2019
Citation: Chen, F., Ma, H., Li, Y., Wang, H., Samad, A., Zhou, J., Zhu, L., Zhang, Y., He, J., Fan, X., Jin, T. 2019. Screening of nanobody specific for peanut major allergen Ara h 3 by phage display. Journal of Agricultural and Food Chemistry. 67:11219-11229. https://doi.org/10.1021/acs.jafc.9b02388.
DOI: https://doi.org/10.1021/acs.jafc.9b02388

Interpretive Summary: Food allergies are caused by adverse immune responses to food proteins, and the past 20 to 30 years marked a rapid rise in the prevalence of food allergy. It was recently estimated that nearly 5% of adults and 8% of children in the US are suffering from food allergies. Peanut allergy has become a major health concern worldwide, and it is the most common cause of fatal food-induced anaphylaxis in the U.S. Despite intensive clinical research within the past decade, a peanut avoidance diet remains the only established approach to the prevention of allergic reactions. Therefore, establishing a reliable method for the detection of peanut allergens is very important and highly sensitive, robust, and low-cost strategy are required. In this study, using a humanized camel nanobody scaffold, a phage display library with randomized antigen binding regions was constructed. By screening the library, a single-domain antibody (named Nb16) specific to the peanut allergen Ara h 3 was obtained. Biochemical characterization and co-crystallization experiments revealed the binding properties of Nb16 to Ara h 3. Additional work is underway to improve the affinity and further explore its application in the food safety industry.

Technical Abstract: 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), co-crystallization, 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 information required to understand the allergenicity of this important peanut allergen. The nanobody Nb16 may have application in the development of biosensors for peanut allergen detection.