Lyophilised pills with phage lytic enzyme 812F1
Staphylococcus aureus is a major causative agent of human and animal diseases. The increasing number of pathogenic strains resistant to antimicrobial drugs is a serious public health problem that can be solved by applications of phage therapy as a suitable alternative to antibiotics treatment. Currently the research focuses on the phage encoded antimicrobial proteins applicable to combat bacterial infections. Endolysins are phage encoded proteins involved in bacteriophage lytic process. Endolysin of phage 812F1 (Twort-like virus) has a modular structure consisting of a C-terminal substrate-binding domain (SH3b) and an N-terminal catalytic domain (CHAP). This enzyme degrades cell wall of staphylococci and it could be used in therapy against staphylococcal bacterial infections.
Compared to classical medical drugs, loosing activity during storage of enzymes could be potential problem in clinical practice. Lyophilisation is probably one of the best methods to conserve enzymatic activity of proteins. Product of classical lyophilisation is a powder without water. Preparation of lyophilised pills is a unique technology whereas lyophilised product keeps its shape. During the preparation of regular pills high pressure and temperature is used. These conditions can influence activity of enzymes. In contrast lyophilisation is optimal for enzymatic activity preservation.
We prepared lyophilized pills that contain phage lytic enzyme (endolysin 812F1). We tested activity of endolysin at plate test and turbidity assay. Both tests show that enzyme keeps its activity after preparation of lyophilised pills. We established a stability study for testing enzymatic activity of pills which are stored at room temperature and at 4 ºC. After three months the enzymatic activity in pills is conserved. We will continue to measure the enzymatic activity for another several months. We assume that lyophilisation can preserve activity of any enzyme, therefore lyophilised pills are potentially an eligible form of medical drug.
Acknowledgment: We kindly acknowledge the financial support from the Technology Agency of the Czech Republic #TA04010038.
Compared to classical medical drugs, loosing activity during storage of enzymes could be potential problem in clinical practice. Lyophilisation is probably one of the best methods to conserve enzymatic activity of proteins. Product of classical lyophilisation is a powder without water. Preparation of lyophilised pills is a unique technology whereas lyophilised product keeps its shape. During the preparation of regular pills high pressure and temperature is used. These conditions can influence activity of enzymes. In contrast lyophilisation is optimal for enzymatic activity preservation.
We prepared lyophilized pills that contain phage lytic enzyme (endolysin 812F1). We tested activity of endolysin at plate test and turbidity assay. Both tests show that enzyme keeps its activity after preparation of lyophilised pills. We established a stability study for testing enzymatic activity of pills which are stored at room temperature and at 4 ºC. After three months the enzymatic activity in pills is conserved. We will continue to measure the enzymatic activity for another several months. We assume that lyophilisation can preserve activity of any enzyme, therefore lyophilised pills are potentially an eligible form of medical drug.
Acknowledgment: We kindly acknowledge the financial support from the Technology Agency of the Czech Republic #TA04010038.
Reference:
Poster Day 4-T12-Pos-11
Session:
Posters Covering the use of viruses to control infection and Processes governing the applied use of viruses
Presenters:
Martin Benesik
Session:
Day 4 Posters Covering: The use of viruses to control infection and Processes governing the applied use of viruses
Presentation type:
Poster presentation
Room:
Poster Halls
Date:
Thursday, 21 July 2016
Time:
12:05 - 15:30