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.;Colgrave, Michelle L.

Shaymaviswanathan Karnaneedi Ludwig Lopata

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.

Journal

Journal of Proteomics

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269

ISBN/ISSN

1876-7737

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Publisher

Elsevier

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DOI

10.1016/j.jprot.2022.104724