Bioprocessing with Salt Active Nucleases - Physiological Conditions

No Other Endonuclease Performs Better at Physiological Conditions

Bioprocessing with Salt Active Nucleases - Physiological Conditions

OverView

Applications

M-SAN HQ - Peak performance where it matters most at physiological conditions

Medium-Salt Active Nuclease High Quality (M-SAN HQ) is a Bioprocessing Grade nuclease developed for removal of both single and double-stranded DNA and RNA at the physiological salt conditions most often used in bioprocessing and biomanufacturing workflows. M-SAN HQ allows you to directly replace Benzonase without changing your workflow.

This novel, nonspecific endonuclease is active over a broad pH range and displays optimum activity at salt concentrations between 125 – 250 mM. Due to its excellent performance at physiological conditions, M-SAN HQ can be used directly in the cell medium or the harvested supernatant without buffer adjustments. This makes M-SAN HQ ideal for the manufacturing of fragile vectors such as lentiviruses and retroviruses.

M-SAN HQ can be directly used in medium without buffer adjustments The high activity of M-SAN HQ at standard cell medium conditions leads to improved DNA clearance compared to other commonly used nucleases. In the data (see Figure 6), an over 2-fold reduction in residual DNA was achieved.

Key Benefits

Compatibility: Ideal for working with both fragile and robust viral vectors and proteins in a variety of cell media
Optimum activity: Optimization for cell media salinity allows both shorter DNA fragments and reduced incubation times.
Cost-Effectiveness: Reduced need for additional reagents and steps can lower production costs
Quality: Maintains the integrity of sensitive or labile biological molecules, ensuring a higher quality end product
Flexibility: Can be used directly in a variety of media without the need for customization

Key Features

High purity (≥ 99%)
No protease detected
Endotoxin-tested
Animal origin-free production
Supplied with extended product documentation

Adapted to use in medium without salinity adjustments

Optimal activity at physiological salinity and pH makes it ideal for DNA removal from mammalian cell media. (see Figures)

The high activity of M-SAN HQ at the physiological salinity and pH found in standard cell medium conditions leads to improved DNA clearance compared to commonly used nucleases.

Simple to Optimize in Cell Media

M-SAN HQ is uniquely formulated to excel in physiological pH conditions, offering high performance at the commonly used cell media pH of 7.4.

While other nucleases often require more alkaline environments, M-SAN HQ stands out for its adaptability and effectiveness in cell media. It's the nuclease that truly aligns with your bioprocessing needs. (See Figure section)

M-SAN HQ ELISA Kit

Toverify removal of M-SAN HQ from bioprocessing workflows, we have developed theM-SAN HQ ELISA kit.

Learn more about the ELISA kit here

Figures

Figure 1. Optimum activity in solutions at physiological salinity

M-SAN High Quality has optimum activity between 125 - 250 mM NaCl, which correlates to the salt content in buffers with physiological salinity. The activity was measured at 37°C in a 25 mM Tris-HCl buffer, pH 7.5, 5 mM MgCl₂ and varying concentrations of NaCl. Maximum activity was set to 100%.

Figure 2. Outperforms commonly used nucleases at physiological salinity

Physiological salinity is equivalent to approx. 150 mM NaCl. At this concentration of salt, M-SAN HQ is up to five-fold more efficient than commonly used nucleases with optimum performance in absence of salt. The activity was measured at 37°C in a 25 mM Tris-HCl buffer, pH 7.2, 5 mM MgCl₂ and varying concentrations of NaCl. The same number of units of each nuclease was used.

Figure 3. Highly compatible with physiological pH

M-SAN HQ has optimal performance in the 7.2 - 8.7 pH range. This allows very high performance in most biological buffers. With a working range starting at pH 6.5, M-SAN HQ is also a candidate for DNA removal from insect cell media. The activity was measured at 37°C in a 25 mM Tris-HCl or Bis-Tris buffer at various pH, 2.5 mM MgCl₂ and 150 mM NaCl. Activity measured at pH 7.7 was set to 100%.

Figure 4. Superior performance at 37°C and above

M-SAN HQ is highly efficient for digesting DNA at 37°C, and tolerate temperatures up to 50°C. The activity was measured in a 25 mM Tris-HCl buffer at pH 7.8 (@measurement temperature), 5 mM MgCl₂ and 150 mM of NaCl. 37°C was set to 100% as this is the temperature used for defining the activity in Units.

Figure 5. Efficient Residual DNA Fragmentation

Gel analysis showing that M-SAN HQ effectively reduces residual host cell DNA to under 8 nucleotides.

Figure 6. M-SAN HQ outperforms other nucleases in removal of host-cell DNA from HEK 293 cells directly in cell media

M-SAN High Quality has optimal activity around physiological conditions, which makes it ideal for DNA removal directly in mammalian cell media In this case, HEK 293 cells were grown in DMEM for 48 hrs before nuclease treatment directly in medium (75 U/ml nuclease). The medium was supplemented with Mg²⁺. to 5 mM. Remaining DNA after 1 hr incubation at 37°C was quantified using Quant-iT™ PicoGreen™ dsDNA Assay Kit.

Properties

Source

Recombinantly produced in Pichia pastoris

Molecular Weight

24.5 kDa

Protein Purity

> 99% by SDS-PAGE analysis

Isoelectric Point

8.62

Unit Definition

One unit is defined as the amount of enzyme that causes a ΔA260 = 1.0 in 30 minutes at 37° in 25 mM Tris-HCl pH 7.6 (@25°C), 2.5 mM MgCl₂, 150 mM NaCl, and 50 μg/ml calf thymus DNA.

Specificity

Nonspecific endonuclease cleaving single and double stranded DNA and RNA.

Working Ranges

Temperature: 20 – 50°C, optimal: 36 – 50°C
Salt concentrations (NaCI): 10 – 500 mM, optimal: 125 mM – 250 mM
Mg2+: > 0.5 mM is required for activity, optimal: 4 – 15 mM
pH: 6.5 – 9.5, optimal: 7.2 – 8.7

Note: The working range is defined as above 10% activity and optimal range as above 80% activity.

Tolerance to Typical Buffer Additives

DTT and other reducing agents may inactivate M-SAN HQ
Urea: Not recommended
EDTA: Not recommended

Quality Control

Recommended Protocols

Ordering Info

Quantity

Product Name

SKU

Pack Size

Concentration

Price

T7 RNA Polymerase

74100-201

10 000 U

50 U/µl

$ 145.00 USD

Bioprocessing with Salt Active Nucleases - Physiological Conditions