Methods and compositions for preserving bacteria
Inventors
Couto, Daniel E. • Reddy, Shilpa • Ulrich, Jil • Michonski, Scott
Assignees
Interested in licensing this patent?
MTEC can help explore whether this patent might be available for licensing for your application.
Publication Number
US-11814615-B2
Publication Date
2023-11-14
Expiration Date
Abstract
The disclosure provides methods and compositions for the preservation of bacteria.
Core Innovation
The patent describes compositions and methods for preserving anaerobic bacteria via lyophilization while maintaining viability, including preservation without substantial loss of viability. The approach uses a stabilization composition that combines a disaccharide lyoprotectant with yeast extract, an antioxidant/reducing component, and a buffer.
The composition includes disaccharide lyoprotectants such as sucrose or trehalose, together with cysteine and sodium metabisulfite and a histidine buffer, to support recovery of preserved bacteria. The patent addresses the problem of preserving anaerobic bacteria so that viability is maintained after lyophilization and over time.
It provides viability assessment using colony forming unit recovery and recovery over time, and it evaluates bacteria subjected to freeze-drying/lyophilization cycles. The document also includes excipient screening, lyophilization cycle temperature ramp rate ranges, preparation of lyophilized solid lyocakes, and scale-up/manufacturing context using freeze-drying trays.
Claims Coverage
The provided claims set includes 1 independent claim, supported by dependent claims that refine viability/recovery, lyophilization temperature ramp rate, and the types of bacteria and composition components.
Preservation composition with disaccharide, yeast extract, cysteine, sodium metabisulfite, and histidine buffer
A method for preserving bacteria that comprises adding bacteria to a composition comprising a disaccharide, yeast extract, cysteine, sodium metabisulfite, and a histidine buffer.
Lyophilization cycle of the bacteria-containing composition
Subjecting the composition comprising the bacteria to a lyophilization cycle.
At least 10% colony forming unit recovery after lyophilization
The method allows for recovery of at least 10% of colony forming units of the bacteria after the lyophilization cycle.
Temperature ramp rate between 0.5°C/min and 3°C/min
The lyophilization cycle comprises one or more steps of a temperature ramp rate between 0.5°C/min to 3°C/min.
Bacteria comprising one or more bacterial strains belonging to the class Clostridia
The bacteria comprise one or more bacterial strains belonging to the class Clostridia.
Trehalose as the disaccharide component
The disaccharide is trehalose.
Measuring colony forming units after lyophilization
The method further comprises measuring the number of colony forming units after subjecting the composition comprising the bacteria to the lyophilization cycle.
Overall, the claim coverage centers on preserving bacteria by combining a disaccharide, yeast extract, cysteine, sodium metabisulfite, and a histidine buffer, followed by lyophilization, with dependent claims refining viability recovery, lyophilization temperature ramp rate, and constraining the bacterial class and disaccharide choice, as well as measuring colony forming units after lyophilization.
Stated Advantages
Allows recovery of at least 10% of colony forming units of the bacteria after the lyophilization cycle.
Documented Applications
Preserving anaerobic bacteria by lyophilization while maintaining viability, including recovery of colony forming units after lyophilization and over time.
Preparation of lyophilized solid lyocakes for preservation of bacteria.
Scale-up/manufacturing context using freeze-drying trays.
Interested in licensing this patent?