Qiagen has filed a patent for a new thermolabile PPR nuclease or its enzymatically active fragment that exhibits high catalyst activity in difficult reaction conditions. The patent also covers the gene encoding the nuclease, a particle of nucleic acid encoding the nuclease, and the method of protein production. The invention has applications in the purification of proteins and viruses, as well as in genetic analyses, gene and cell therapies, and therapeutic protein purification. GlobalData’s report on Qiagen gives a 360-degree view of the company including its patenting strategy. Buy the report here.
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According to GlobalData’s company profile on Qiagen, Nucleoside chemical synthesis was a key innovation area identified from patents. Qiagen's grant share as of September 2023 was 47%. Grant share is based on the ratio of number of grants to total number of patents.
Thermolabile ppr nuclease for purification of proteins and viruses
A recently filed patent (Publication Number: US20230287369A1) describes an isolated recombinant protein that includes a PPR nuclease or a fragment of it. The protein sequence consists of amino acids 42 to 269 of SEQ ID NO: 2 and includes a hexa-HIS purification tag (SEQ ID NO: 3B). This recombinant protein can be irreversibly inactivated by incubating it at 52°C for 15 minutes in the presence of 1-5 mM DTT or at a temperature of 52°C or lower for longer than 15 minutes in the presence of 1-5 mM DTT.
The enzymatic activity of the PPR nuclease is maintained in various salt concentrations. It remains active in NaCl concentrations ranging from 0 to 1400 mM, MgCl concentrations ranging from 5 to 200 mM, urea concentrations ranging from 0 to 6000 mM, ammonium sulfate concentrations ranging from 0 to 200 mM, and imidazole concentrations ranging from 0 to 400 mM.
The patent also includes an expression plasmid called pD454-PPR-AmpR (SEQ ID NO: 4), which contains a nucleic acid sequence encoding the isolated recombinant protein. This nucleic acid sequence includes a T7 phage promoter or another promoter that is active in an E. coli expression system. A recombinant Escherichia coli strain of AmpR can be transformed using this expression plasmid.
Methods for producing the PPR nuclease protein are described in the patent. This involves culturing the recombinant strain of Escherichia coli containing the expression vector pD454-PPR-AmpR on a medium, inducing PPR nuclease gene expression by adding IPTG, and isolating and purifying the PPR nuclease protein.
The patent also covers methods for isolating and purifying the PPR nuclease or its enzymatically active fragment. This involves expressing the PPR nuclease or its fragment in a host cell, followed by separation of the PPR nuclease from the host cell and/or cell culture medium.
The isolated recombinant protein comprising the PPR nuclease or its enzymatically active fragment has various applications. It can be used for the purification of recombinant proteins, reducing DNA content in recombinant proteins, decontaminating reagents and reaction mixtures for genetic analyses, purifying virus vectors, exosomes, pharmaceutical grade proteins, enzymes, antibodies, or vaccination antigens, and removing nucleic acid contamination in culture media for mammalian fermentation and microbiological processes.
The patent also mentions the use of the isolated recombinant protein for virus vector purification in gene and cell therapies or chimeric antigen receptor (CAR) T-cell immunotherapy.
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GlobalData Patent Analytics tracks bibliographic data, legal events data, point in time patent ownerships, and backward and forward citations from global patenting offices. Textual analysis and official patent classifications are used to group patents into key thematic areas and link them to specific companies
