Screening of specific binding peptide for ß-lactoglobulin using phage display technology.

ß-lactoglobulin (ß-Lg) is a major food allergen, there is an urgent need to develop a rapid method for detecting ß-Lg in order to avoid contact or ingestion by allergic patients. Peptide aptamers have high affinity, specificity, and stability, and have broad prospects in the field of rapid detection. Using ß-Lg as the target, this study screened 11 peptides (P1-11) from a phage display library. Using molecular docking technology to predict binding energy and binding mode of proteins and peptides. Select the peptides with the best binding ability to ß-Lg (P5, P7, P8) through ELISA. Combining them with whey protein, casein, and bovine serum protein, it was found that P7 has the best specificity for ß-Lg, with an inhibition rate of 87.99%. Verified by molecular dynamics that P7 binds well with ß-Lg. Therefore, this peptide can be used for the recognition of ß-Lg, becoming a new recognition element for detecting ß-Lg.

Effect of ultrahigh-pressure treatment on the structure and allergenicity of peach allergenic proteins.

Peach is a common plant-derived allergenic food and ultrahigh-pressure treatment is often used in peach products. In our study, an in-depth analysis of the structural and allergenicity changes of peach allergenic proteins after UHP treatment was performed by spectroscopy, mass spectrometry combined with serology and cytology. The results indicated that UHP treatment could reduce the content of peach soluble proteins and cause changes in secondary and tertiary structures. In addition, more hydrophobic residues were exposed and proteins tended to polymerize after UHP-treatment. The results of immunological assays showed that UHP treatment could reduce the IgE binding capacity of peach proteins and affect the ability of basophil degranulation, the upregulation of some cytokines may contribute to the reduction of peach protein allergenicity. Notably, UHP treatment may lead to the masking of some digestion sites in Pru p 3 epitopes, thus impeding human digestion and increasing the potential risk of allergenicity.

Effect of Roasting on the Conformational Structure and IgE Binding of Sesame Allergens.

Sesame can trigger a systemic allergic reaction. In the present study, we investigated the responses of the structure and IgE binding of sesame allergens to different roasting treatments (120, 150, and 180 °C for 5 to 30 min). We analyzed the tryptic digestion peptides using a label-free mass spectrometry method. The total amount of soluble proteins in sesame was significantly reduced by roasting at 180 °C, followed by 150 °C. Ses i 1 was the most stable protein during processing as it still possessed a higher protein abundance compared to other allergens after roasting under 180 °C. The most unstable allergens were Ses i 4 and Ses i 7, which suffered severe protein degradation at 180 °C. Roasting at 180 °C remarkably increased the secondary structure content of α-helices but decreased that of ß-sheets, whereas roasting at 120 and 150 °C had a limited effect on the secondary structure of sesame proteins. Moreover, serum pool Western blot analysis showed that the main allergens were oleosin of Ses i 4 and Ses i 5. The IgE-binding ability of sesame allergens was significantly decreased under 180 °C roasting, as well as the solubility of sesame proteins, which showed remarkable congruence in changes. Relative quantification results indicate that individual sesame allergens respond differently to the roasting process. In general, sesame allergens are unstable under roasting treatment. Therefore, the allergenic potential of sesame allergens may be minimized by selecting appropriate parameters during processing.

Identification of Major B-Cell Linear Epitopes of Peach Allergen Pru p 3 Using Immune Slot-Blot Microarray Assay.

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.