{"id":11564,"date":"2025-03-17T17:20:47","date_gmt":"2025-03-17T17:20:47","guid":{"rendered":"https:\/\/www.caltagmedsystems.co.uk\/information\/?p=11564"},"modified":"2025-03-18T15:53:43","modified_gmt":"2025-03-18T15:53:43","slug":"endonuclease-maxnuclease-gmp-grade","status":"publish","type":"post","link":"https:\/\/www.caltagmedsystems.co.uk\/information\/endonuclease-maxnuclease-gmp-grade\/","title":{"rendered":"KACTUS: Endonuclease – MaxNuclease\u2122 GMP-Grade"},"content":{"rendered":"\n

Background<\/h2>\n\n\n\n

With the rapid development of the biopharmaceutical field, the variety of biological products is increasing. At the same time, biological products have strict quality control systems, with residual nucleic acids being a key focus for regulatory agencies both domestically and internationally. Products such as AAV vectors, antibody drugs, vaccines, and recombinant protein drugs are produced using continuously passaged cells, and despite meticulous purification processes, residual host nucleic acids may still be present. These residual nucleic acids can pose risks such as pathogenicity and tumourigenicity. Hence regulatory agencies have strict guidelines for the amount of residual nucleic acids in biological products.<\/p>\n\n\n\n

In addition, in the CAR-T cell therapy industry, large-scale purification of lentiviruses, protein extraction, and other fields also require strict control of nucleic acid residues. Currently, methods for removing nucleic acids mainly include enzymatic degradation, polyethyleneimine (PEI) precipitation, and ion exchange chromatography. Among these, enzymatic methods using nonspecific nucleases are highly favored for their efficiency, cost-effectiveness, and ease of use.<\/p>\n\n\n\n

GMP Endonuclease to Degrade all DNA & RNA<\/h2>\n\n\n\n

KACTUS has developed a GMP-grade universal nuclease – MaxNuclease\u2122, which degrades all DNA and RNA into 2-5 base oligonucleotides. It has been developed and validated according to GMP manufacturing standards to ensure the traceability of raw materials throughout the production process. This product has been registered with the U.S. FDA Drug Master Files (#036799), providing documentation on aspects such as production processes and control, and material control.<\/p>\n\n\n\n

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MaxNuclease\u2122, GMP<\/strong><\/a><\/div>\n<\/div>\n\n\n\n

High-efficiency, enzymatic nucleic acid degradation for biologics manufacturing.<\/h2>\n\n\n\n
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What is MaxNuclease\u2122?<\/h4>\n\n\n\n

How does MaxNuclease\u2122 work?<\/h4>\n\n\n\n

What is MaxNuclease\u2122 good for?<\/h4>\n\n\n\n

MaxNuclease\u2122 is identified from Serratia <\/em>marcescens and is genetically engineered and expressed in E. coli <\/em>under cGMP manufacturing standards. MaxNuclease\u2122 is a non-specific nuclease with high activity and specificity that degrades all forms of nucleic acids including single- and double-stranded, linear and circular nucleic acids.<\/p>\n\n\n\n

MaxNuclease\u2122 is a homodimer of two 30 kDa subunits containing two disulfide bonds that are essential for activity and stability. It hydrolyses internal phosphodiester bonds between nucleotides in nucleic acids to produce 5′-monophosphate oligonucleotides of 2-5 bases in length.<\/p>\n\n\n\n

Products such as lentiviral or AAV vectors, antibody drugs, vaccines, and recombinant protein drugs are expressed and produced by continuously passaged cells. Even after a fine purification process, host nucleic acids may remain in the products, and the residual nucleic acids may cause pathogenicity, tumourigenesis, etc. Residual nucleic acids need to be removed to a safe level in the final drug product.<\/p>\n<\/div>\n\n\n\n

Product Features<\/h2>\n\n\n\n
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Manufactured in a GMP-compliant facility<\/p>\n\n\n\n

Raw materials free from animal-derived components<\/p>\n\n\n\n

Low Endotoxin: \u2264 0.01 EU\/kU<\/p>\n\n\n\n

Strict quality management for clinical manufacturing<\/p>\n\n\n\n

FDA Drug Master Files: DMF #036799<\/p>\n<\/div>\n\n\n\n

Product Specifications & Quality<\/h2>\n\n\n\n

Product Specifications<\/h4>\n\n\n\n
Parameter<\/th>Specification<\/th><\/tr><\/thead>
Catalog No.<\/td>GMP-NUC-SE101<\/td><\/tr>
FDA Drug Master Files<\/td>#36799<\/td><\/tr>
Source<\/td>E. coli with endonuclease gene from Serratia marcescens<\/td><\/tr>
Molecular Weight<\/td>Approximately 27.8 kDa<\/td><\/tr>
Formulation<\/td>20mM Tris-HCl, 20mM NaCl, 2mM MgCl2, 50% Glycerol, pH 8.0<\/td><\/tr>
Storage<\/td>Store at -20\u00b15\u00b0C. Avoid repeated freeze-thaw.<\/td><\/tr>
Activity<\/td>\u2265250 U\/\u00b5L analysed by degradation of Herring Sperm DNA<\/td><\/tr>
Unit Definition<\/td>One unit corresponds to the amount of enzyme required to produce a change in absorbance at 260 nm of 1.0 in 30 minutes, at 37\u00b0C and pH 8.0.<\/td><\/tr><\/tbody><\/table><\/figure>\n\n\n\n

Quality Control Criteria<\/h4>\n\n\n\n
Assay<\/th>Specification<\/th><\/tr><\/thead>
Activity (Dissolve herring sperm DNA)<\/td>\u2265 250 U\/\u00b5L<\/td><\/tr>
Purity (Bis-Tris)<\/td>\u2265 95%<\/td><\/tr>
Purity (SEC-HPLC)<\/td>\u2265 99%<\/td><\/tr>
Residual Protease<\/td>Negative<\/td><\/tr>
Residual Host Protein<\/td>\u2264 10 ppm<\/td><\/tr>
Endotoxin<\/td>\u2264 0.01 EU\/kU<\/td><\/tr>
Sterility<\/td>Negative<\/td><\/tr>
Residual Heavy Metal<\/td>\u2264 10 ppm<\/td><\/tr>
Mycoplasma<\/td>Negative<\/td><\/tr><\/tbody><\/table><\/figure>\n\n\n\n

Product Validation Data<\/h2>\n\n\n\n
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Host Cell DNA Removal<\/h3>\n\n\n\n

Plasmid DNA Removal<\/h3>\n\n\n\n

Degradation of PCR Product, Genomic DNA, and Plasmid DNA<\/h3>\n\n\n\n

Virus harvest solution was treated with 25U\/mL and 50U\/mL endonuclease at 37\u00b0C for 2 hours, respectively. Detection of Host Cell DNA (HCD) residue was analysed. MaxNuclease\u2122 has higher degradation activity versus Competitor B demonstrated by lower HCD residue for both 25U\/mL and 50U\/mL working concentrations.<\/p>\n\n\n\n

Virus harvest solution was treated with 25U\/mL and 50U\/mL endonuclease at 37\u00b0C for 2 hours, respectively. Detection of plasmid DNA (pDNA) residue was analysed. MaxNuclease\u2122 has higher degradation activity versus Competitor B demonstrated by lower pDNA residue for both 25U\/mL and 50U\/mL working concentrations.<\/p>\n\n\n\n

MaxNuclease\u2122 added to PCR product, genomic DNA, and plasmid DNA shows comparable degradation activity of nucleic acids to leading competitors.<\/p>\n<\/div>\n\n\n\n

GMP Compliance<\/h2>\n\n\n\n

KACTUS’ MaxNuclease\u2122 has been developed and verified with reference to cGMP production standards. The manufacturing process is in accordance with GMP production standards to ensure the traceability of raw materials in the production process. At present, this product has also been filed with the U.S. FDA Drug Master Files (DMF #036799).<\/p>\n\n\n\n

MaxNuclease\u2122 is available in long-term bulk supply with batch-to-batch consistency to ensure suitability for industrial applications.<\/p>\n\n\n\n

Quality Management System<\/h3>\n\n\n\n