Diverting the Use of Hand-Operated Tablet Press Machines to Bioassays: A Novel Protocol to Test ‘Waste’ Insoluble Shell Matrices
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Published:2024-04-01
Issue:2
Volume:7
Page:30
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ISSN:2409-9279
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Container-title:Methods and Protocols
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language:en
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Short-container-title:MPs
Author:
Lutet-Toti Camille12ORCID, Da Silva Feliciano Marie1, Debrosse Nelly1, Thomas Jérôme1ORCID, Plasseraud Laurent3ORCID, Marin Frédéric1ORCID
Affiliation:
1. UMR CNRS-uB-EPHE 6282 ‘Biogéosciences’, Université de Bourgogne, 21000 Dijon, France 2. Dipartimento di Chimica “Giacomo Ciamician”, Alma Mater Studiorum Università di Bologna, 40126 Bologna, Italy 3. Institut de Chimie Moléculaire de l’Université de Bourgogne, ICMUB UMR CNRS-uB 6302, 21000 Dijon, France
Abstract
To mineralize their shells, molluscs secrete a complex cocktail of proteins—collectively defined as the calcifying shell matrix—that remains occluded in the exoskeleton. Nowadays, protein extracts from shells are recognized as a potential source of bioactive substances, among which signalling molecules, bactericides or protease inhibitors offer the most tangible perspectives in applied sciences, health, and aquaculture. However, one technical obstacle in testing the activity of shell extracts lies in their high insolubility. In this paper, we present a protocol that circumvents this impediment. After an adapted shell protein extraction and the production of two organic fractions—one soluble, one insoluble—we employ a hand-operated tablet press machine to generate well-calibrated tablets composed of 100% insoluble shell matrix. FT-IR monitoring of the quality of the tablets shows that the pressure used in the press machine does not impair the molecular properties of the insoluble extracts. The produced tablets can be directly tested in different biological assays, such as the bactericidal inhibition zone assay in Petri dish, as illustrated here. Diverting the use of the hand-operated tablet press opens new perspectives in the analysis of insoluble shell matrices, for discovering novel bioactive components.
Funder
Université de Bourgogne Universtà di Bologna Alma Mater Studiorum AFFDU Société Française de Biologie des Tissus Minéralisés Fondation Arthritis CNRS Observatoire des Sciences de l’Univers
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