YEASEN UCF. ME ® UltraNuclease — Your BEST choice for degrading nucleic acids

1. What is totipotent nuclease?
2. What are the usage scenarios of totipotent nuclease?
3. What are the advantages that totipotent nuclease needs to have?
4. FAQ
5. Related Products

1. What is totipotent nuclease?

Totipotent nucleases are genetically modified endonucleases with the ability to degrade a wide range of nucleic acid forms, encompassing single-stranded, double-stranded, linear, native, and denatured nucleic acids. They generate 5'-monophosphate oligonucleotides with lengths ranging from 3 to 5 bases and exhibit no specific base recognition. These versatile nucleases exhibit robust stability and digestion activity across various conditions, making them highly efficient for eliminating nucleic acid residues in samples or products. This enhances sample purity and the biological effectiveness of products, rendering totipotent nucleases the preferred enzyme preparation for scientific research, vaccines, and industries such as protein and polysaccharide pharmaceuticals.

Furthermore, totipotent nucleases play a crucial role in the removal of nucleic acids during the preparation of vaccines, antibodies, cell therapy, and other biological products, ensuring compliance with FDA regulations stipulating that residual nucleic acid in the final product for therapeutic use must not exceed 10 pg/dose. By degrading nucleic acids, totipotent nucleases reduce the viscosity of cell lysates, facilitating the solution filtration/ultrafiltration process during the purification of cell-derived particles such as viruses, AAV vectors, and inclusion bodies. This streamlines processing, improves centrifugation, enhances the separation efficiency of precipitation and supernatant, and boosts the effectiveness of chromatographic purification, ultimately increasing yield and product purity.

2. What are the usage scenarios of totipotent nuclease?

UltraNuclease has a wide range of applications and here are the specific usage scenarios.

2.1 Virus purification (lentivirus, AAV, recombinant adenovirus vaccine, inactivated virus, oncolytic virus, etc.)

The utilization of viruses within the biomedical sector is expanding significantly, spanning applications such as vaccines, cell therapy, gene therapy, and various other fields. Host cells responsible for virus production are predominantly continuous passage cells (CCLs). CCLs exhibit a genetic predisposition for unregulated growth, granting them the ability to proliferate indefinitely. However, the presence of residual DNA from CCLs can pose potential risks, such as uncontrolled human cell proliferation leading to tumor formation. Furthermore, these residual DNA fragments may contain infectious viral genomes, raising concerns about infection risk alongside tumorigenesis. Consequently, stringent control measures are imperative for managing the residual DNA in biological products, ensuring their safety. In this regard, our UCF.ME™ UltraNuclease offers a secure solution for virus purification.

Salt plays a pivotal role in mitigating protein or virus aggregation, facilitating the separation of DNA and RNA from proteins and other cellular components. Salt Active UltraNuclease, being a nonspecific endonuclease, exhibits optimal activity at high salt concentrations. In high-salt conditions, enzymes gain improved access to released DNA and RNA, enhancing their degradation. Nucleases with heightened activity in high-salt environments prove more effective in enhancing purification processes. Salt Active UltraNuclease can be employed to reduce viscosity in cell supernatants and lysates, enhancing purification efficiency under high salt conditions. This enzyme also reduces host nucleic acid residues to the picogram level, thereby enhancing the performance and safety of various biological product applications, including virus purification, vaccine manufacturing, and the production of protein and polysaccharide pharmaceuticals. 

2.2 Purification of the recombinant protein expressed in E. coli

Certain proteins expressed in E. coli have a tendency to aggregate into inclusion bodies, complicating the purification process as they often become intertwined with the host DNA. The resultant ruptured bacterial cells can exhibit high viscosity, significantly diminishing protein purification efficiency. However, by introducing 10-50 U/mL of UCF.ME™UltraNuclease to the bacterial solution during ultrasonic disruption and incubating the sample at either 37°C for 30 minutes or 4°C overnight following the completion of disruption, viscosity in the bacterial solution can be effectively reduced. This, in turn, substantially enhances the efficiency of the purification process. In summary, you can rely on our UCF.ME™ UltraNuclease to facilitate the effective purification of the protein.

2.3 Prevent cell clumping

In recent years, the use of cryopreserved peripheral blood mononuclear cells (PBMC) in immunoassays has increased dramatically. The biggest characteristic of PBMC cells is that they are very easy to clump after resuscitation, which can easily lead to low cell quality and unreliable test results. When the cells are resuscitated, the appropriate concentration (25-50 U/mL) of UCF.ME™UltraNuclease is added to the culture medium and the cells are co-cultured, which can effectively prevent cell clumping. Hence, you can make the most of our UCF.ME™ UltraNuclease to increase the odds of a successful outcome.

3. What are the advantages that totipotent nuclease needs to have?

3.1 Wide range of applications

UCF.ME™ UltraNuclease can cut and degrade various forms of DNA and RNA under a variety of experimental conditions and can be applied to remove residual nucleic acid in various biological products.

3.2 High purity, high enzyme activity

For our UCF.ME™ UltraNuclease, its enzyme purity is greater or equal to 99% (HPLC), and its enzyme activity is greater or equal to1.5*106 U/mg (using an absolute quantitative method for general substrate).

3.3 Strong adaptability

UCF.ME™ UltraNuclease has high stability in storage and freeze-thaw. We store UCF.ME™ UltraNuclease at -20 °C, 4 °C, and 37 °C to detect the trend of enzyme activity. When placed at 37 °C for 2 weeks, there was no significant change in enzyme activity. Meanwhile, to ensure the stability of the enzyme, we use a combination of dry ice + ice pack to transport UCF.ME™ UltraNuclease to your place. It also has strong tolerance and can adapt to a variety of operating conditions.

3.4 Comply with Pharmacopoeia

UCF.ME™ UltraNuclease is manufactured in UCF.ME (Ultra Clean Factory. Molecular Enzyme) is a kind of Ultra-clean enzyme. What is an Ultra-clean enzyme? It is a molecular enzyme that achieves quality standards of no HCD、HCR、HCP, DNASE FREE、RNASE FREE, no protease residues, no microbial residues, no pathogens, animal origin, antibiotic pollution, ultra-low endotoxin Residual, high enzyme activity, and high purity, etc. Hence, you can trust our products, safely and reliably. 

3.5 GMP production

The whole process of UltraNuclease production is completed in the GMP production workshop, and the quality inspection plan is strictly by the pharmacopeia method, meeting the large-scale demand from research and development to production.

UCF.ME™ UltraNuclease’s partner —— UltraNuclease ELISA kit
In normal purification steps, UCF.ME™ UltraNuclease is easily removed as an impurity.  To detect the residue of UCF.ME™ UltraNuclease, YEASEN has developed the supporting UltraNuclease ELISA kit, which can accurately detect the residue of UCF.ME™ UltraNuclease in the sample. The kit has excellent linearity, repeatability, recovery rate, and specificity.

UltraNuclease ELISA kit   

The linear range is between 0.047 and 3 ng/mL, and R2 is greater than 0.99. with Good inter-batch reproducibility, we add a fixed concentration of UltraNuclease to the sample and use 3 different batches of ELISA kits for detection. The detection recovery rate is between 70% and 110% when the nuclease concentration is between 3 and 0.186 ng/mL, and CV is less than 10%. What’s more, the simulated sample recovery rates of UltraNuclease with different concentrations are all between 70% and 130% and are better than other brands. It also has strong specificity that we evaluated the non-specific binding of the UltraNuclease antibody to 8 common samples. The results of the interference group and the control group overlapped, and no interference was seen.

Following UltraNuclease's DMF filing, YEASEN has obtained DMFs of salt active ultranuclease from the USA FDA.

4. FAQ

Q1: In which step to add UltraNuclease?

A: It depends on the samples you are dealing with. In virus purification, it is generally added after virus clarification; in E. coli protein purification, it can be added when the cells are lysed; in dealing with cell clumping, it can be directly added to the medium for co-cultivation with cells.

Q2: If the reaction temperature cannot reach 37°C, how to use it?

A: The enzyme activity of UltraNuclease is affected by temperature, treatment time, and enzyme activity units. If the temperature cannot reach 37°C, the amount of enzyme can be added appropriately, or the incubation time can be extended.

Q3: Is UltraNuclease compatible with protease inhibitors?

A: Compatible, but be aware that many protease inhibitors contain EDTA. When EDTA starts to be higher than 1mM, EDTA will inhibit the activity of some nucleases.

Q4: How to remove UltraNuclease?

A: It can be removed by methods such as TFF, dialysis, and chromatographic columns.

Q5: Can the UltraNuclease ELISA kit be used for the detection of other brands of totipotent nuclease residues?

A: We do not recommend doing this. The ELISA kit is completely developed based on UltraNuclease and can accurately quantify UltraNuclease. Altipotent nucleases of other brands may be different from UltraNuclease in terms of sequence and production process, and the test results may be inaccurate.

 5. Related Products

The products provided by Yeasen are as follows.     

Table 1 Related Products