Comparison of protein extraction protocols and allergen mapping from black soldier fly Hermetia illucens
Bose, Utpal, Broadbent, James A., Juhász, Angéla, Karnaneedi, Shaymaviswanathan, Johnston, Elecia B., Stockwell, Sally, Byrne, Keren, Limviphuvadh, Vachiranee, Maurer-Stroh, Sebastian, Lopata, Andreas L., and Colgrave, Michelle L. (2022) Comparison of protein extraction protocols and allergen mapping from black soldier fly Hermetia illucens. Journal of Proteomics, 269. 104724.
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Abstract
Exploration of important insect proteins — including allergens — and proteomes can be limited by protein extraction buffer selection and the complexity of the proteome. Herein, LC-MS/MS-based proteomics experiments were used to assess the protein extraction efficiencies for a suite of extraction buffers and the effect of ingredient processing on proteome and allergen detection. Discovery proteomics revealed that SDS-based buffer yields the maximum number of protein groups from three types of BSF samples. Bioinformatic analysis revealed that buffer composition and ingredient processing could influence allergen detection. Upon applying multi-level filtering criteria, 33 putative allergens were detected by comparing the detected BSF proteins to sequences from public allergen protein databases. A targeted LC-MRM-MS assay was developed for the pan-allergen tropomyosin and used to assess the influence of buffer composition and ingredient processing using peptide abundance measurements.
Item ID: | 76385 |
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Item Type: | Article (Research - C1) |
ISSN: | 1876-7737 |
Keywords: | Allergen, Black soldier fly, Extraction protocol, Insect, LC-MRM-MS, LC-MS/MS, Proteomics, Tropomyosin |
Copyright Information: | © 2022 The Authors. Published by Elsevier B.V. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/bync-nd/4.0/). |
Date Deposited: | 23 Mar 2023 01:15 |
FoR Codes: | 31 BIOLOGICAL SCIENCES > 3102 Bioinformatics and computational biology > 310205 Proteomics and metabolomics @ 50% 32 BIOMEDICAL AND CLINICAL SCIENCES > 3205 Medical biochemistry and metabolomics > 320506 Medical biochemistry - proteins and peptides (incl. medical proteomics) @ 50% |
SEO Codes: | 10 ANIMAL PRODUCTION AND ANIMAL PRIMARY PRODUCTS > 1099 Other animal production and animal primary products > 109999 Other animal production and animal primary products not elsewhere classified @ 100% |
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