Faecal Microbiota Lyophilization with Cryoprotectant Buffer
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Solution Overview
Problem
Current faecal microbiota transplantation methods are empirical and lack effective preservation of anaerobic bacteria, leading to variable efficacy and the need for repeated treatments, especially for conditions like Clostridium difficile infections, with a lack of stable and viable samples on an industrial scale.
Innovation Solution
A method involving mixing faecal microbiota with a diluent containing polyols, di- to pentasaccharides, and maltodextrins, followed by freezing at temperatures below -50°C and lyophilization, to create a stable lyophilizate that preserves bacterial viability and diversity.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If faecal microbiota transplantation is performed using conventional empirical methods, then the treatment can be administered, but the viability of anaerobic bacteria is not effectively preserved, leading to variable efficacy
Solution Approach 1:
The patent applies preliminary action by preparing the faecal microbiota sample in advance with specific protective measures before transplantation. The sample is mixed with a buffer solution containing cryoprotectants (glycerol, trehalose, sucrose) and antibiotics, then frozen at controlled temperatures to preserve bacterial viability before the actual transplantation procedure.
Solution Approach 2:
The patent changes physical and chemical parameters to improve bacterial preservation. Specifically, it controls freezing temperature (−80°C), adjusts buffer composition (cryoprotectant concentrations), and modifies storage conditions to maintain anaerobic environment and prevent bacterial death during storage and transplantation.
2Reliability
If repeated treatments are administered to ensure efficacy, then the desired therapeutic effect may be achieved, but the treatment duration and complexity increase
Solution Approach 1:
The patent prepares the microbiota sample in advance with optimized buffer composition and freezing conditions to maximize bacterial survival. This preliminary preparation ensures that a single transplantation contains sufficient viable bacteria to achieve therapeutic effect, reducing the need for repeated treatments.
3Productivity
If industrial scale production of faecal microbiota samples is implemented, then availability increases, but maintaining bacterial viability and diversity becomes more difficult
Solution Approach 1:
The patent develops a universal buffer solution formulation and freezing protocol that can be applied to multiple samples simultaneously at industrial scale. The buffer contains cryoprotectants and antibiotics that work together to preserve bacterial viability, and the standardized procedure ensures consistent results across large numbers of samples.
Solution Approach 2:
The patent optimizes physical parameters for industrial-scale processing, including freezing temperature (−80°C), buffer composition ratios, and storage conditions. These parameter changes ensure that bacterial viability and diversity are maintained even when processing large volumes of samples in an industrial setting.
Applied Scientific Principles
This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.
Function Achieved in This Case
The method produces a stable and effective lyophilizate that maintains bacterial viability and diversity, enhancing the efficacy of faecal microbiota transplantation and providing a long-term solution for intestinal dysbioses, including Clostridium difficile infections.
Implementation Method 1
mixing a sample of faecal microbiota from a donor subject with a diluent chosen from polyols, di- to pentasaccharides, maltodextrins and mixtures thereof
Implementation Method 2
freezing the mixture obtained in A) at a temperature less than −50° C., preferably comprised between −70° C. and −100° C.
Implementation Method 3
freezing the mixture obtained in A) at a temperature less than −50° C., preferably comprised between −70° C. and −100° C., then lyophilizing it
Data Source
AI summary
Method for lyophilisation of a simple of fecal microbiota. The present invention relates to a method for lyophilisation of a sample of fecal microbiota from a donor subject, comprising the following steps: A) mixing of a sample of fecal microbiota from a donor subject with a diluent selected from polyols, disaccharides to pentasaccharides, maltodextrins and mixtures thereof, and B) freezing the mixture obtained in A) at a temperature of less than −50° C., preferably of between −70° C. and −100° C., followed by the lyophilisation thereof.