2.1 Allergen amino acid sequence retrieval
The amino acid sequences of
P01070 (trypsin inhibitor), P04347 (Gly m 6.0501), P04776 (Gly m
6.0101), P05046 (lectin), P11827 (Gly m 5.0201), P25974 (Gly m 5.03),
and P26987 (Gly m 4) were downloaded from the Uniprot database
(https://www.uniprot.org/), and the detailed information of the
seven soy allergens are shown in Table 1.
2.2 T cell
epitope prediction
The “MHC-II Binding Predictions” tool in the IEDB (Immune Epitope
Database Analysis Resource) database was used to predict the soybean
allergen peptides that could bind to HLA class II molecules [21].
The potential sequence was submitted to the software in Fasta format,
and the IEDB recommended method, combining with the consensus method and
the NetMHCIIpan method, was selected for prediction, and thereinto, the
consensus method considers the combination of any three of four methods,
including artificial neural network (ANN) alignment method,
stabilization matrix (SMM) alignment method, combinatorial library
method, and Sturniol method. A total of 27 HLA molecules (15 HLA-DR
molecules, 6 HLA-DQ molecules, and 6 HLA-DP molecules) were used to
predict T cell epitopes, and the epitope length parameter was set to 15
amino acids as a suggestion.