Salt Active Nuclease
High Quality (Bioprocessing grade)

SAN High Quality (Bioprocessing grade) is the ultimate solution for efficient removal of nucleic acids in manufacturing and bioprocessing workflows. This nonspecific, recombinant endonuclease has optimum activity at high salt concentrations, which can improve efficiency and yield in various workflows.

Salt is an important component of various purification schemes. The presence of salt can minimize aggregation, increase target solubility and improve target yield. High salt enables contaminating DNA to dissociate from associated proteins and become available for degradation. SAN High Quality is highly compatible with the use of high salt conditions.

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High activity at high salt conditions

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Supplied with extended product documentation

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Compatible with SAN HQ ELISA

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High purity (≥ 98%)

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No protease detected

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Active at low temperatures

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Figure 1: Optimum activity in solutions with high salinity. SAN High Quality has optimum activity between 400 - 650 mM NaCl but can be used in a broad range of NaCl-concentrations. The activity was measured at 37°C in a 25 mM Tris-HCl buffer, pH 8.5, 10 mM MgCl2 with varying concentrations of NaCl. Maximum activity was set to 100%. Similar results were obtained using KCl instead of NaCl (results not shown).
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Figure 2: Optimum performance at slightly alkaline conditions. SAN HQ performs best in the 8.2 - 8.8 pH range. A working range starting at pH 7.3 allows use in most biological buffers. The activity was measured at 37°C in a 25 mM Tris-HCl or Bis-Tris Propane buffer buffer at various pH, 5 mM MgCl2 and 500 mM NaCl. Maximum activity was set to 100%.
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Figure 3: Suitable for DNA removal in the ambient to 37°C range. SAN HQ originates from a cold-adapted organism, which allows excellent performance at the temperature ranges commonly used in bioprocessing. For temperature sensitive products, DNA digestion overnight at 4°C is also possible. The activity was measured in a 25 mM Tris-HCl buffer at pH 8.5 (@ measurement temperature), 5 mM MgCl2 and 500 mM NaCl. 37°C was set to 100% as this is the temperature used for defining the activity in Units.
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Figure 4: Robust performance at 37°C. SAN HQ shows excellent stability during use at 37°C, which is the typical incubation temperature in bioprocessing. After incubation at relevant DNA digestion conditions for 1 hour at 37°C, no significant loss of activity was observed compared to control sample kept on ice in enzyme dilution buffer. Buffer compositions: Lysis buffer (50 mM Tris-HCl pH 8.0 @ 25°C, 5 mM MgCl2, 0.5 % Triton X-100, 500 mM NaCl), DMEM (DMEM supplemented with 10 % FBS, 5 mM MgCl2 and 300 mM NaCl), Lysis buffer in DMEM (Lysis buffer and DMEM 1:1).

Properties

  • Source: Recombinantly produced in Pichia pastoris
  • Molecular weight: The protein is glycosylated. Protein size without glycosylation is 26 kDa.
  • Protein purity: > 98% by SDS-PAGE analysis
  • Isoelectric point: 9.55
  • Unit definition: One unit is defined as the amount of enzyme that causes a ΔA260 = 1.0 in 30 minutes at 37°C in 25 mM Tris-HCI  pH 8.5 (@25°C), 5 mM MgCl2, 500 mM NaCl, and 50 µg/ml calf thymus DNA.
  • Specificity: Nonspecific endonuclease cleaving single- and double-stranded DNA and RNA.
  • Working ranges:
    • Temperature: 7 – 38°C, 4°C overnight, optimal: 30 – 38°C
    • Salt concentrations (NaCl / KCl): 100 mM – 900 mM, optimal: 400 - 650 mM
    • Mg2+: > 1 mM is required for activity, optimal: 5 – 50 mM
    • pH: 7.3 – 9.2, optimal: 8.2 - 8.8

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

  • Tolerance to typical buffer additives:
    • Imidazole: 20% activity at 350 mM Imidazole
    • Glycerol: 20% activity at 35% glycerol
    • Triton X-100: No reduction in activity (tested up to 15%)
    • SDS: Not recommended
    • Urea: Not recommended
    • Reducing agents (e.g. DTT, TCEP): will result in inactivation

SAN HQ ELISA kit

ArcticZymes offers SAN HQ ELISA kit to confirm the removal of SAN High Quality (Bioprocessing Grade) in bioprocessing and biomanufacturing applications.

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Publications

SAN-HQ Applications

  1. Moving from the bench towards a large scale, industrial platform process for adeno-associated viral vector purification.
    Adams B, Bak H, Tustian AD.
    Biotechnology & Bioengineering. 2020; 117 (10): 3199-3211.

  2. Cytosine and adenine base editing of the brain, liver, retina, heart and skeletal muscle of mice via adeno-associated viruses.
    Levy JM, Yeh WH, Pendse N, Davis JR, Hennessey E, Butcher R, Koblan LW, Comander J, Liu Q, Liu DR.
    Nature Biomedical Engineering. 2020; 4(1): 97-110.

  3. Global Representations of Goal-Directed Behavior in Distinct Cell Types of Mouse Neocortex.
    Allen WE, Kauvar IV, Chen MZ, Richman EB, Yang SJ, Chan K, Gradinaru V, Deverman BE, Luo L, Deisseroth.
    Neuron. 2017; 94 (4): 891-907.

  4. Identification of peripheral neural circuits that regulate heart rate using optogenetic and viral vector strategies.
    Rajendran PS, Challis RC, Fowlkes CC, Hanna P, Tompkins JD, Jordan MC, Hiyari S, Gabris-Weber BA, Greenbaum A, Chan KY, Deverman BE, Münzberg H, Ardell JL, Salama G, Gradinaru V, Shivkumar K.
    Nat Commun. 2019; 10: 1944.

  5. Multiplexed peroxidase-based electron microscopy labeling enables simultaneous visualization of multiple cell types.
    Zhang Q, Lee WA, Paul DL, Ginty DD.
    Nat Neurosci. 2019; 22: 828–839.

  6. Near physiological spectral selectivity of cochlear optogenetics.
    Dieter A, Duque-Afonso CJ, Rankovic V, Jeschke M, Moser T.
    Nat Commun. 2019; 10: 1962.

  7. Mining, analyzing, and integrating viral signals from metagenomic data.
    Zheng T, Li J, Ni Y, Kang K, Misiakou MA, Imamovic L, Chow BKC, Rode AA, Bytzer P, Sommer M, Panagiotou G.
    Microbiome. 2019; 7: 42.

  8. Structures of the Human PGD2 Receptor CRTH2 Reveal Novel Mechanisms for Ligand Recognition.
    Wang L, Yao D, Krishna Deepak RNV, Liu H, Xiao Q, Fan H, Gong W, Wei Z, Zhang C.
    Molecular Cell. 2018; 72, 48-59.

  9. Ultrafast optogenetic stimulation of the auditory pathway by targeting-optimized Chronos. 
    Keppeler D, Merino RM, Lopez de la Morena D, Bali B, Huet AT, Gehrt A, Wrobel C, Subramanian S, Dombrowski T, Wolf F, Rankovic V, Neef A, Moser T.
    EMBO J. 2018; 37(24): e99649.

  10. Engineered AAVs for efficient noninvasive gene delivery to the central and peripheral nervous systems. 
    Chan K, Jang M, Yoo B, Greenbaum A, Ravi N, Wu WL, Sánchez-Guardado L, Mazmanian SK, Deverman BE, Gradinaru V.
    Nat Neurosci. 2017; 20: 1172–1179.

  11. Global Representations of Goal-Directed Behavior in Distinct Cell Types of Mouse Neocortex.
    Allen WE, Kauvar IV, Chen MZ, Richman EB, Yang SJ, Chan K, Gradinaru V, Deverman BE, Luo L, Deisseroth K.
    Neuron. 2017; 94 (4): 891-907.e6.