SAN HQ 2.0

SAN High Quality 2.0 is the second generation of our SAN High Quality enzyme. SAN HQ 2.0 offers wider compatibility with downstream processes used in bioprocessing workflows while exhibiting the same salt tolerance and other biochemical characteristics as SAN HQ.

The downstream compatibility improvements are of particular importance if used with anionic exchange media or using size exclusion chromatography / TFF below 100 kDa pore size.
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High activity at high salt conditions

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High purity
(≥ 99%)

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Easier removal

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Compatible with SAN HQ 2.0 ELISA

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Active at low temperatures

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Detergent free

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No protease detected

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Table 1. Easy separation. Due to its high isoelectric point (9.55), SAN HQ 2.0 binds strongly to cation exchange media, even at elevated pH and salt concentrations, where few other proteins binds. SAN HQ 2.0 does not typically bind to anion exchange media. SAN HQ 2.0 does not have a His-tag, and consequently does not bind to IMAC media. This allows easy separation if the protein of interest contains a His-tag.

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Figure 1: Optimum activity in solutions with high salinity. SAN High Quality 2.0 has optimum activity between 400 - 650 mM NaCl but can be used in a broad range of NaCl-concentrations, near identical to the characterisitics of SAN HQ (pale green). 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 2.0 performs best in the 8.2 - 8.8 pH range. A working range starting at pH 7.3 allows use in most biological buffers. SAN HQ 2.0 demonstrate close to indentical pH tolerance as SAN HQ (pale green). The activity was measured at 37°C in a 25 mM Tris-HCl or Bis-Tris Propane 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 2.0 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 temperature tolerance is near identical to that of SAN HQ (pale green ). The activity was measured in a 25 mM Tris-HCl buffer at pH 8.5 (@ measurement temperature), 5 mM MgCl2 and 500 mM of 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 2.0 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: 26 kDa
  • Protein purity: > 99% 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. The enzyme degrades DNA vs RNA in a 10:1 ratio.
  • Working ranges:
    • Temperature: 7 – 38°C, 4°C overnight, optimal: 30 – 38°C
    • Salt concentration (NaCl / KCl): 100 - 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 ~10% of activity and optimal range is ~80% of activity.

  • Tolerance to typical buffer additive:
    • Triton X-100: No reduction in activity (tested up to 15%)

SAN HQ 2.0 ELISA kit

ArcticZymes offers SAN HQ 2.0 ELISA kit to confirm the removal of SAN HQ 2.0 in bioprocessing and biomanufacturing applications.

Key features

  • Sensitive
    • Quantification range: 0.2 – 12.8 ng/ml
  • Precise
    • RSD ≤ 15%
  • Accurate
    • 100% ± 15%
Download SAN HQ 2.0 brochure
Download SAN HQ 2.0 ELISA kit leaflet