# Diverting the Use of Hand-Operated Tablet Press Machines to Bioassays: A Novel Protocol to Test ‘Waste’ Insoluble Shell Matrices

## TL;DR

This paper introduces a new method to test insoluble proteins from mollusc shells using tablet press machines, enabling the discovery of bioactive substances.

## Contribution

A novel protocol using tablet press machines to test insoluble shell matrices, overcoming their high insolubility for bioassays.

## Key findings

- A hand-operated tablet press machine can produce calibrated tablets from 100% insoluble shell matrix.
- FT-IR analysis confirms that tablet pressing does not damage the molecular properties of the shell extracts.
- The tablets can be directly used in biological assays, such as bactericidal inhibition zone tests.

## 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.