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 |
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.
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
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.
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.
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.
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.