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Electrochemical Protease Biosensor for Milk Safety Control

Periodic Reporting for period 1 - EPROMILK (Electrochemical Protease Biosensor for Milk Safety Control)

Reporting period: 2021-10-04 to 2023-10-03

The European Union (EU) is a substantial producer of milk and dairy products. Milk production takes place in all EU Countries and represents a significant portion of the value of EU’s agricultural output, being estimated at around 15 million tons per year. Food safety of products produced in or imported into Europe is ensured due to strict European regulations. Dairy industries carry out processes that ensure the quality and safety of their products such as UHT processing. Although UHT milk is bacteriologically stable for months at room temperature, its shelf-life is often compromised by age gelation initiated by the proteolytic activity originated by proteases. The effect of thermostable proteases, resistant to the heating conditions in UHT processing on dairy products, especially on UHT milk quality, is drawing increasing attention worldwide. Residual protease activity can lead to non-desirable effects in milk such as high viscosity, sedimentation, age gelation, fat separation and bitterness, reducing consumer acceptability.
The aim of EPROMILK has been to evaluate and demonstrate the feasibility of developing a fast and simple method based on electrochemical biosensors that can be applied for on-site protease activity for the dairy sector.
The implementation of EPROMILK has resulted in the definition of a novel biosensing strategy for protease detection in dairy products at the point of need in a fast and reliable way.
EPROMILK has fostered working on the development and optimisation of two biosensing strategies for the electrochemical detection of milk proteases. Proteases are enzymes that cleave amide bonds in aminoacid chains, and their natural activity is used as principle of detection in these strategies.
A simple biosensing strategy for plasmin detection in milk has been developed and its applicability confirmed for plasmin detection in milk. The prototype will be verified and validated with a higher number of milk samples and further optimisation performed, if required. The prototype will be fully adapted for being run by the BIO7000 portable device.
Furthermore, EPROMILK has enabled the acquisition of deep understanding of the use of novel biorecognition elements for biosensing applications and obtaining an ink catalogue for screen printing purposes. Both results will be integrated into BIOLAN´s R&D know-how for further application within the company.
The work carried out within EPROMILK has been broadly disseminated by taking part in various dissemination, communication and outreach events targeting both the scientific community and the general public.
Nevertheless, results have not been fully disseminated, as they have been considered as industrial secret so far, until a full patentability evaluation is performed.
The biosensing strategy proposed constitutes a novel strategy for plasmin and other proteases detection as it reduces device-to-device and batch-to-batch irreproducibility typically derived from strategies that need for surface modification.
The implementation of EPROMILK by the ER and the hosting institution, has delivered an electrochemical biosensing strategy for protease detection in milk with the potential to translate after further optimisation into a first in-kind portable, and easy to use analytical tool using disposable consumables for the dairy industry. This will be a unique platform in the market, constituting a highly disruptive tool for the dairy sector enabling a fast decision-making at the point of need. There is no doubt that the use of this tool by the dairy industry will translate into a positive impact on consumers and therefore strengthening their position in the market.
IOLAN´s BIO 7000 portable measuring device and a disposable test strip
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