Nucleases are enzymes that cleave phosphodiester bonds of DNA or RNA. These enzymes are involved in a variety of biological processes, including DNA replication, removal of DNA damage, repair of DNA double-strand breaks, synthesis of active messenger RNA, processing of ribosomal and transfer RNAs, and gene regulation. Nucleases include DNases, RNases, RNase H, endonucleases, exonucleases, and more. DNase degrades DNA, and RNase degrades RNA. RNase H is the nuclease that degrades the RNA strand of RNA/DNA hybrid molecules. Endonuclease is responsible for cleaving double-stranded DNA at internal phosphodiester bonds, and exonuclease removes nucleotides sequentially from the 5′- or 3′-end of the polynucleotide chain. Some nucleases have both exonuclease and endonuclease activities. Some nucleases cleave only single-stranded or double-stranded nucleic acids, but some can cleave both single-stranded and double-stranded nucleic acids. A variety of nucleases are used for biotechnology applications such as restriction digestion, degradation of selected nucleic acids, and trimming the ends of RNA and DNA polymers.
Salt Active Nucleases
During bioprocessing, the primary role of nucleases is to efficiently digest and fragment extraneous genetic material into small pieces to facilitate its removal during downstream processing. While most nucleases can efficiently degrade naked DNA into tiny fragments under optimal conditions, realistic biological processing is more complex. The DNA that needs to be removed often exists as chromatin, embedded in a complex matrix containing remnants of the lysed host cell and large amounts of products. High salt concentration enables DNA to dissociate from proteins and become available for degradation. Therefore, the ideal nuclease could efficiently segment DNA under more complex, high-salt conditions.
Amerigo Scientific offers salt active nucleases that exhibit high activity at a wide range of salt concentrations, making them ideal for removing nucleic acid impurities in manufacturing and bioprocessing workflows to improve the efficiency and yield. Medium-Salt Active Nuclease High Quality (M-SAN HQ) is a nuclease suitable for the isotonic/physiological salt range in cell media. Salt Active Nuclease High Quality (SAN HQ) is developed for removing nucleic acids at high salt conditions. M-SAN HQ and SAN HQ are nonspecific endonucleases that cleave single and double stranded DNA and RNA. These nucleases can be used for the removal of nucleic acids during protein production, vaccine manufacturing, and viral vector preparation.
Working Ranges
| |
M-SAN HQ |
SAN HQ |
HL-SAN |
| Temperature |
20-50°C, optimal: 36-50°C |
5-38°C, 4°C overnight, optimal: 32-38°C |
7-38°C and 4°C overnight, optimal: 30-38°C |
| Salt Concentrations |
(NaCI): 10-500 mM, optimal: 125 mM-250 mM |
(NaCl / KCl): 0-1000 mM, optimal: 400-700 mM |
(NaCI / KCI): 0-1000 mM, optimal: 400-700 mM |
| Mg2+ |
> 0.5 mM is required for activity, optimal: 4-15 mM
|
>1 mM is required for activity, optimal: 6-50 mM |
>1 mM is required for activity, optimal: 5-50 mM |
| pH |
6.5-9.5, optimal: 7.2-8.7 |
7.0-9.5, optimal: 8.0-8.8 |
6.8-9.5, optimal: 8.0-8.9 |
Note: The working range is defined as above 10% activity and optimal range as above 80% activity.
The residual amount of nucleases in biological products is one of the important indicators to measure the quality of biological products. Therefore, we offer SAN HQ ELISA kit and M-SAN HQ ELISA kit that utilize a classical sandwich ELISA methodology to detect and quantify residual levels of the nuclease of choice, ensuring that final products and in-process samples meet stringent quality standards.
GMP-grade Nucleases
Amerigo Scientific offers GMP-grade nucleases that are manufactured under the relevant guidelines for Good Manufacturing Practice (GMP). The FDA Drug Master File (DMF) LOA is available upon request.
BenzoNuclease® is an endonuclease that degrades nucleic acids, whether DNA and RNA are single-stranded, double-stranded, linear, circular, or supercoiled. By BenzoNuclease®, all free nucleic acids present in solution are digested and reduced to 5 '-monophosphoride-terminated oligonucleotides 3 to 5 bases in length. We also offer BenzoNuclease® ELISA Kit with high sensitivity and specificity for the detection and quantification of BenzoNuclease® residues. In addition, we offer the GMP-grade Cas9 nuclease expressed in E. coli which is cultured in animal-free medium.
Working Ranges
| |
BenzoNuclease®, GMP Grade* |
SAN HQ, GMP Grade** |
| Temperature |
0 - 42°C, optimal: 37°C |
5-38°C, 4°C overnight, optimal: 32-38°C |
| Salt concentrations |
(Na+, K+, etc.): 0-150 mM, optimal: 0-20 mM |
(NaCl / KCl): 0-1000 mM, optimal: 400-700 mM |
| Mg2+ |
1-10 mM is required for activity, optimal: 1-2 mM |
>1 mM is required for activity, optimal: 6-50 mM |
| pH |
6.0-10.0, optimal: 8.0 - 9.2 |
7.0-9.5, optimal: 8.0-8.8 |
* The working range is defined as above 15% activity and optimal range as above 90% activity.
** The working range is defined as above 10% activity and optimal range as above 80% activity.
Related Products
The class 2 CRISPR-Cas enzymes are RNA-guided endonucleases that cleave target DNA/RNA sequences. Amerigo Scientific offers Cas9, Cas12a, Cas12b, Cas12f, Cas13 nucleases from a variety of biological origins.
The production of highly biologically active mRNA often depends on reliable design and preparation, in which high-quality mRNA-associated enzymes are essential. Amerigo Scientific provides a range of FDA DMF filed enzymes to support the development and manufacture of mRNA therapeutics.
.
