Document Type

Journal Article

Publication Title

Journal of Proteomics

Volume

269

PubMed ID

36096435

Publisher

Elsevier

School

School of Science

Funders

1st Singapore-Australia Bilateral Program on Innovations in Food for Precision Health, 2019 (Grant number: SG-AUS2019_191D4) / Commonwealth Scientific and Industrial Research Organisation (Australia) / James Cook University (Australia and Singapore) / Agency for Science, Technology and Research (A*STAR, Singapore)

Comments

Bose, U., Broadbent, J. A., Juhász, A., Karnaneedi, S., Johnston, E. B., Stockwell, S., ... & Colgrave, M. L. (2022). Comparison of protein extraction protocols and allergen mapping from black soldier fly Hermetia illucens. Journal of Proteomics, 269, Article 104724. https://doi.org/10.1016/j.jprot.2022.104724

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. Significance: We demonstrated that the selection of protein extraction buffer and the processing method could influence protein yield and cross-reactive allergen detection from processed and un-processed black soldier fly (BSF) samples. In total, 33 putative allergens were detected by comparing the detected BSF proteins to sequences from public allergen protein databases. An LC-MRM-MS assay was developed for tropomyosin, indicating the importance of buffer selection and processing conditions to reduce BSF samples' allergenicity.

DOI

10.1016/j.jprot.2022.104724

Creative Commons License

Creative Commons Attribution-Noncommercial-No Derivative Works 4.0 License
This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 4.0 License.

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