GMP-Grade MaxNuclease™

MaxNuclease, a broad-spectrum nuclease, is derived from Serratia Marcescen. It is expressed by genetically engineered E.coli and purified, all in a GMP-Grade environment. This enzyme can degrade all forms of DNA and RNA, including single-stranded, double-stranded, linear, circular, native, and denatured nucleic acids. It digests them into 5'-monophosphate oligonucleotides of 3-5 bases in length, and has no base recognition specificity. MaxNuclease can maintain high stability and enzyme activity under a wide range of conditions and is suitable for removing nucleic acid residues in samples and improving product purity in pharmaceutical industries such as viral vaccines, viral vectors, and recombinant proteins.

1) Removing DNA/RNA from biological products. The US FDA requires that the nucleic acid content of each dose of biological products for therapeutic use be less than 10pg. MaxNuclease can be used for the removal of nucleic acids in industrial biological products such as vaccines, polysaccharides, and proteins, so that the final nucleic acid content of biological products can meet regulatory requirements and the efficacy of biological products can be improved.

2) Purifying cell culture-derived products. Nucleic acids easily adhere to the surface of cell-generated particles such as virus-like particles (VLP), virus particles, and inclusion bodies, which changes the size or charge of the particles and causes the aggregation of these particles, such as during the storage of peripheral blood single cells (PBMC) agglomeration phenomenon. MaxNuclease can effectively degrade nucleic acid, avoid the influence of nucleic acids on cell products and purification, and help improve the purification efficiency of cell products.

3) Reducing the viscosity of lysed cells. MaxNuclease can degrade all forms of nucleic acid, reduce the viscosity of cell lysate, increase protein yield, improve separation effect, make it easy to filter (especially ultrafiltration), and facilitate downstream chromatographic purification operations.

4) Preparing samples for biochemical analysis. In analyses such as ELISA, chromatography, two-phase electrophoresis, and footprinting analysis, treatment of protein samples containing nucleic acids with MaxNuclease can improve resolution and improve recovery.

KACTUS' MaxNuclease detection kit uses the double-antibody sandwich method to determine the content of MaxNuclease. Both the detection antibody and the coating antibody are recombinantly expressed monoclonal antibodies. The detection antibody is pre-coupled with HRP, which simplifies the operation steps and saves time. The kit has a sensitivity of 23.44 pg/mL and a linear range of 46.88 pg/mL-3000.00 pg/mL. 

We have tested the stability experiments at 25°C and 37°C. It was found that MaxNuclease can still maintain >90% of the enzyme activity under these two conditions for 7 days, which proves that the enzyme is very stable in a suitable storage buffer. Contact us for stability data.

Store at -20°C. Avoid repeated freezing and thawing. The enzyme is valid for at least 2 years. We recommend shipping on dry ice.

MaxNuclease can be used for a variety of purposes (see “What are the main applications of MaxNuclease?”). Its usage and dosage vary according to the purpose of use. For the production of biological products such as viral vaccines, viral vectors, recombinant proteins, etc., MaxNuclease is generally added after harvesting and before purification.

This depends on your application scenario and experimental condition. Generally, 25-50U/mL is suitable in the cell lysis step of an AAV production process. Due to different buffers and application scenarios, there will be relatively large differences in enzyme activity and optimal concentrations. For example, if the temperature is lower than 37°C during use, the enzyme activity will decrease. Compared to 37°C, MaxNuclease maintains about 22% of the enzyme activity at 4°C. When the temperature is lower than 37°C, the same nucleic acid removal effect can be achieved by appropriately extending the reaction time. Temperature, nuclease concentration, and time are the three conditions that mainly affect enzyme activity, and these three aspects can be optimized in practical applications. If you would like assistance in the usage of MaxNuclease, please contact us at support@kactusbio.us. 

1-2mM Mg2+ is necessary to maintain enzyme activity. The reaction system can be adjusted by adding additional Mg2+ to maintain enzyme activity. 

Monovalent cations can inhibit enzyme activity, such as >300mM Na+, K+, or >100mM ammonium sulfate precipitation agent. Additionally, >1mM EDTA will inhibit enzyme activity because EDTA will chelate Mg2+ ions necessary for enzyme activity. Denaturing agents such as urea, as well as proteases, will also inactivate the enzyme.

MaxNuclease is easily removed in downstream purification with techniques such as depth filtration and tangential flow filtration (TFF). In pharmaceutical manufacturing, endonucleases are often removed by ion exchange chromatography. The PI of MaxNuclease is 6.85, and anion exchange chromatography is generally used to make the MaxNuclease flow through or be eluted.

In addition, the residue of MaxNuclelase is also part of the quality control analysis of pharmaceutical products. Generally, an ELISA kit can be used to detect MaxNuclease (including active and inactive) remaining in the product. The MaxNuclease detection kit developed by KACTUS uses a double antibody sandwich method to accurately quantify the residue of MaxNuclease in the product, with a sensitivity of 23.44 pg/mL and a linear range of 46.88 pg/mL-3000.00 pg/mL.

One unit of activity corresponds to the amount of enzyme required to change the absorbance at 260 nm by 1.0 under the condition of excess salmon sperm DNA at 37°C and pH 8.0 for 30 minutes (equivalent to the amount of enzyme required to completely digest 37ug of DNA substrate). The verified enzyme activity of MaxNuclease is ≥ 250 U/µL. 

Yes, MaxNuclease is manufactured in our dedicated GMP-Grade production facility and undergoes strict quality release testing. Additionally, we provide customizable documentation for regulatory filings. MaxNuclease has also been submitted to the FDA Drug Master Files (DMF #036799). 

Yes, with our prokaryotic fermentation system and supernatant expression, there is no need for renaturation, which allows us to avoid mixing intermediate renaturation products. Additionally, high expression levels allow for large-scale individual batches to meet manufacturing production requirements.

Documents such as product specification, COA, COO, MSDS, TSE/BSE declaration can be provided. This product has passed the FDA DMF Type II record, with DMF #036799. Please contact support@kactusbio.us for more information.