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Identification of Major B-Cell Linear Epitopes of Peach Allergen Pru p 3 Using Immune Slot-Blot Microarray Assay
Journal of Agricultural and Food Chemistry ( IF 6.1 ) Pub Date : 2022-06-24 , DOI: 10.1021/acs.jafc.2c01448
Wenhan Kang 1, 2 , Jiukai Zhang 2 , Hong Li 3 , Ning Yu 2 , Rui Tang 3 , Xiulan Sun 1 , Lei He 2 , Jinlyu Sun 3 , Ying Chen 1, 2
Affiliation  

Pru p 3, one of the most representative proteins of the lipid transfer proteins (LTPs), is responsible for clinical allergic reactions to food of peach origin. The identification of Pru p 3 epitopes is not comprehensive due to different methods and principles of epitope screening. In addition, evaluation of the stability of the epitopes and the validation of the immunological key amino acids still need further research. Therefore, in the present study, an immune slot-blot microarray assay was performed to screen the epitopes from Pru p 3 overlapping peptide library, and a new epitope (P-1, AA1-16, ITCGQVSSALAPCIPY) was identified and two identified epitopes were deeply investigated (P-2, AA12-27, PCIPYVRGGGAVPPAC; P-3, AA23-38, VPPACCNGIRNVNNLA). The stability of these epitopes was then verified by thermal processing treatment and digestion experiments. Moreover, the key amino acids of the three identified epitopes were obtained by epitope amino acid mutation combined with slot-blot experiments. These findings may contribute to the further understanding of Pru p 3 and the prevention of peach allergy.

中文翻译:

使用免疫 Slot-Blot 微阵列检测鉴定桃过敏原 Pru p 3 的主要 B 细胞线性表位

Pru p 3 是脂质转移蛋白 (LTP) 中最具代表性的蛋白质之一,负责对桃源性食物的临床过敏反应。由于表位筛选的方法和原则不同,Pru p 3表位的鉴定并不全面。此外,表位稳定性的评价和免疫学关键氨基酸的验证仍需进一步研究。因此,在本研究中,进行免疫槽印迹微阵列检测以从 Pru p 3 重叠肽库中筛选表位,并鉴定了一个新的表位(P-1、AA1-16、ITCGQVSSALAPCIPY)和两个已鉴定的表位。深入研究(P-2,AA12-27,PCIPYVRGGGAVPPAC;P-3,AA23-38,VPPACCNGIRNVNNLA)。然后通过热处理和消化实验验证这些表位的稳定性。此外,通过表位氨基酸突变结合槽印迹实验获得了三个鉴定表位的关键氨基酸。这些发现可能有助于进一步了解 Pru p 3 和预防桃过敏。
更新日期:2022-06-24
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