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Benz-Neburase™ GMP, His-tag

Removal of plasmid and host residual DNA during Lentivirus (LV) production

Step 1: Dilute Benz-Neburase to 10 kU/ml and place it in a chromatography refrigerator at 4 °C for later use.
Step 2: Mix the harvested cell suspension (5 ml) and add 10 μl Benz-Neburase, mix thoroughly and place in a 37 °C water bath for 60 min.
Step 3: After the incubation, centrifuge at 1300 g for 10 min to remove the cells and cell debris.
Step 4: After the centrifugation, measure the HCD, plasma residues in the samples, and Fluorescence activated Cell Sorting (FACS) can be used to determine the functional titer of lentivirus particles.

The test results show that Benz-Neburase can remove DNA and plasmid residue in virus production process more effectively than the competitor product. The use of Benz-Neburase has minimal impact on the viral production recovery. The recovery data further indicates that the GenScript Benz-Neburase outperforms the competitor product.

Benz-Neburase™ GMP, His-tag

Removal of plasmid and host residual DNA during adeno associated virus (AAV) production

Step 1: After harvesting cell suspension, break up the cells, then add 100 U Benz-Neburase to 2 ml cell suspension, mix thoroughly and place in a 37°C water bath for 60 min.
Step 2: After the incubation, centrifuge to remove the cells and cell debris at 1600 g for 10 min.
Step 3: After the centrifugation, measure the HCD and plasmid DNA residues in the samples, and Viral genome (Vg) recovery was used to quantify the adeno associated viral production.

The test results show that Benz-Neburase can also remove DNA and plasmid residue in AAV virus production process more effectively than the competitor product. The use of Benz-Neburase has minimal impact on the viral production. The data further indicates that the GenScript Benz-Neburase outperforms the competitor product

Benz-Neburase™ GMP, His-tag

Reduction of the viscosity of bacterial lysate

Step 1: Centrifuge the bacterial culture, remove the supernatant, and then add the lysate.
Step 2: Treat the sample with Benz-Neburase at a final concentration of 2.5 U/ml, incubate at 37 °C for 30 min.
Step 3: Centrifuge to observe the viscosity of the precipitate and supernatant.

The test results show that Benz-Neburase can greatly reduce the viscosity of bacterial lysate.

Benz-Neburase™ GMP, His-tag

Prevention of cell clumping

Step 1: Spread the adhered cells in a 24-well plate and treat them with control buffer (left) and 50 U/ml Benz-Neburase (right) at 37 °C for 30 min.
Step 2: Observe the cells using a microscope

The test result show that Benz-Neburase can efficiently reduce cell clumping.

Benz-Neburase™ GMP, His-tag

Prevention of cell clumping

Step 1: Treat SUP-T1 cells (left) and NK-92 cells (right) with 2 μl of Benz-Neburase at different concentrations (20-500 U/ml).
Step 2: Incubate overnight in an incubator at 37 °C in 5% CO2.

The test results indicate Benz-Neburase has minimal to no impact on the cell viability.

Benz-Neburase™ GMP, His-tag

Lane M:DNA marker
Lane 1:PCR Product
Lane 2:Benz-Neburase + PCR Product
Lane 3:Competitor endonuclease + PCR product
Lane 4:Genomic DNA
Lane 5:Benz-Neburase + Genomic DNA
Lane 6:Competitor endonuclease + Genomic DNA
Lane 7:Plasmid DNA
Lane 8:Benz-Neburase + Plasmid DNA
Lane 9:Competitor endonuclease + Plasmid DNA
Lane 10:RNA
Lane 11:Benz-Neburase + RNA
Lane 12:Competitor endonuclease + RNA

In 20 μl reaction volume, use 1U of Benz-Neburase to digest different kinds of nucleic acid at 37 °C for 10 min.

The test results show that Benz-Neburase is effective in digesting various forms of DNA and RNA, with efficiency identical to competing products.

Benz-Neburase™ GMP, His-tag

GenScript’s offerings are cost-effective with reliable and proven performance for removal of contaminating nucleic acids. Furthermore, the Benz-Neburase’s offerings are produced under stringent quality controls with materials and manufacturing process that are fully traceable, allowing for support of IND filing and progression of development stages for various applications.
Z03627
¥1800

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Description

Benz-Neburase is a genetically engineered endonuclease from Serratia marcescens used in the purification processes of biological molecules and biopharmaceutical production such as vaccine, viral vector, gene and cell therapy manufacturing facilities. The enzyme is highly effective, nonspecific endonuclease capable of digesting all forms of DNA and RNA into 3-5 bp nucleotides.  
Benz-Neburase is active in a broad range of conditions and is free of proteolytic activity, making the enzyme especially useful for applications with challenging DNA contaminants, such as lysed host cells in viral vector manufacturing processes. The nuclease requires divalent cation, preferably Mg2+ for activity, displays a broad pH tolerance, ranging from pH 6 to pH 10, optimal at 8-8.5, and has a wide temperature tolerance, ranging from 35 °C to 55 °C. Benz-Neburase is a physiologic homodimer and functions more progressively than the monomer. Two disulfide bonds in the nuclease are crucial to its activity and stability.   
GenScript’s offerings are cost-effective with reliable and proven performance for removal of contaminating nucleic acids. Furthermore, the Benz-Neburase’s offerings are produced under stringent quality controls with materials and manufacturing process that are fully traceable, allowing for support of IND filing and progression of development stages for various applications. 

Note

For research and manufacturing use. Direct human use, including taking orally and injection are forbidden.

Product Introduction Benz-Neburase is a genetically engineered endonuclease from Serratia marcescens used in the purification processes of biological molecules and biopharmaceutical production such as vaccine, viral vector, gene and cell therapy manufacturing facilities. The enzyme is highly effective, nonspecific endonuclease capable of digesting all forms of DNA and RNA into 3-5 bp nucleotides. 
Benz-Neburase is active in a broad range of conditions and is free of proteolytic activity, making the enzyme especially useful for applications with challenging DNA contaminants, such as lysed host cells in viral vector manufacturing processes. The nuclease requires divalent cation, preferably Mg2+ for activity, displays a broad pH tolerance, ranging from pH 6 to pH 10, optimal at 8-8.5, and has a wide temperature tolerance, ranging from 35 °C to 55 °C. Benz-Neburase is a physiologic homodimer and functions more progressively than the monomer. Two disulfide bonds in the nuclease are crucial to its activity and stability. 
GenScript’s offerings are cost-effective with reliable and proven performance for removal of contaminating nucleic acids. Furthermore, the Benz-Neburase’s offerings are produced under stringent quality controls with materials and manufacturing process that are fully traceable, allowing for support of IND filing and progression of development stages for various applications. 

Expression System E.coli
Species Serratia Marcescen
Tag N-terminal 6 × His
Theoretical Molecular Weight 27.5 kDa
Biological Activity ≥ 1.1 × 106 U/mg
One unit of Benz-Neburase™ GMP, His-tag is defined as the amount of enzyme that causes a ∆A260 of 1.0 (equivalent to the complete digestion of 37 μg DNA) in 30 min.
Formulation Supplied as a solution of 20 mM Tris-HCl, 2 mM MgCl2, 20 mM NaCl, 50% Glycerol, pH 8.0.
Storage & Stability This product remains stable for up to 2 weeks at 4 °C or up to 30 months at -20 °C.
Avoid repeated freeze-thaw cycles.
Do not store below -20 °C!

This enzyme requires 1-2 mM Mg2+ to be used to remove all forms of DNA and RNA, including double stranded, single stranded, linearized, and circular forms.

Appearance Clear, colorless liquid
Purity ≥ 99% as analyzed by SEC-HPLC
≥ 95% as analyzed by SDS-PAGE
Enzyme Activity ≥ 250 U/μl
Endotoxin Level ≤ 0.01 EU/kU
Residual HCP ≤ 10 µg/mg
Protease Activity Non-detectable
Bioburden < 1 CFU/ml
Heavy Metal Residue ≤ 10 ppm
Mycoplasma Negative

  • Benz-Neburase™ GMP, His-tag
  • Benz-Neburase™ GMP, His-tag

    Removal of plasmid and host residual DNA during Lentivirus (LV) production

    Step 1: Dilute Benz-Neburase to 10 kU/ml and place it in a chromatography refrigerator at 4 °C for later use.
    Step 2: Mix the harvested cell suspension (5 ml) and add 10 μl Benz-Neburase, mix thoroughly and place in a 37 °C water bath for 60 min.
    Step 3: After the incubation, centrifuge at 1300 g for 10 min to remove the cells and cell debris.
    Step 4: After the centrifugation, measure the HCD, plasma residues in the samples, and Fluorescence activated Cell Sorting (FACS) can be used to determine the functional titer of lentivirus particles.

    The test results show that Benz-Neburase can remove DNA and plasmid residue in virus production process more effectively than the competitor product. The use of Benz-Neburase has minimal impact on the viral production recovery. The recovery data further indicates that the GenScript Benz-Neburase outperforms the competitor product.

  • Benz-Neburase™ GMP, His-tag
  • Benz-Neburase™ GMP, His-tag

    Removal of plasmid and host residual DNA during adeno associated virus (AAV) production

    Step 1: After harvesting cell suspension, break up the cells, then add 100 U Benz-Neburase to 2 ml cell suspension, mix thoroughly and place in a 37°C water bath for 60 min.
    Step 2: After the incubation, centrifuge to remove the cells and cell debris at 1600 g for 10 min.
    Step 3: After the centrifugation, measure the HCD and plasmid DNA residues in the samples, and Viral genome (Vg) recovery was used to quantify the adeno associated viral production.

    The test results show that Benz-Neburase can also remove DNA and plasmid residue in AAV virus production process more effectively than the competitor product. The use of Benz-Neburase has minimal impact on the viral production. The data further indicates that the GenScript Benz-Neburase outperforms the competitor product

  • Benz-Neburase™ GMP, His-tag
  • Benz-Neburase™ GMP, His-tag

    Reduction of the viscosity of bacterial lysate

    Step 1: Centrifuge the bacterial culture, remove the supernatant, and then add the lysate.
    Step 2: Treat the sample with Benz-Neburase at a final concentration of 2.5 U/ml, incubate at 37 °C for 30 min.
    Step 3: Centrifuge to observe the viscosity of the precipitate and supernatant.

    The test results show that Benz-Neburase can greatly reduce the viscosity of bacterial lysate.

  • Benz-Neburase™ GMP, His-tag
  • Benz-Neburase™ GMP, His-tag

    Prevention of cell clumping

    Step 1: Spread the adhered cells in a 24-well plate and treat them with control buffer (left) and 50 U/ml Benz-Neburase (right) at 37 °C for 30 min.
    Step 2: Observe the cells using a microscope

    The test result show that Benz-Neburase can efficiently reduce cell clumping.

  • Benz-Neburase™ GMP, His-tag
  • Benz-Neburase™ GMP, His-tag

    Prevention of cell clumping

    Step 1: Treat SUP-T1 cells (left) and NK-92 cells (right) with 2 μl of Benz-Neburase at different concentrations (20-500 U/ml).
    Step 2: Incubate overnight in an incubator at 37 °C in 5% CO2.

    The test results indicate Benz-Neburase has minimal to no impact on the cell viability.

  • Benz-Neburase™ GMP, His-tag
  • Benz-Neburase™ GMP, His-tag

    Lane M:DNA marker
    Lane 1:PCR Product
    Lane 2:Benz-Neburase + PCR Product
    Lane 3:Competitor endonuclease + PCR product
    Lane 4:Genomic DNA
    Lane 5:Benz-Neburase + Genomic DNA
    Lane 6:Competitor endonuclease + Genomic DNA
    Lane 7:Plasmid DNA
    Lane 8:Benz-Neburase + Plasmid DNA
    Lane 9:Competitor endonuclease + Plasmid DNA
    Lane 10:RNA
    Lane 11:Benz-Neburase + RNA
    Lane 12:Competitor endonuclease + RNA

    In 20 μl reaction volume, use 1U of Benz-Neburase to digest different kinds of nucleic acid at 37 °C for 10 min.

    The test results show that Benz-Neburase is effective in digesting various forms of DNA and RNA, with efficiency identical to competing products.


For laboratory research use only. Direct human use, including taking orally and injection and clinical use are forbidden.


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