UltraNuclease (Benzonase Nuclease) – The total Solution for Nucleic Acid Residue removing in Biologics

In the production and process flow of therapeutic drugs (such as antibodies, viral vector drugs, etc.), the removal of nucleic acid impurities is crucial. UltraNuclease, also known as broad-spectrum nuclease or non-restrictive endonuclease, can cleave and degrade various forms of DNA and RNA under multiple experimental conditions, and is widely used for removing nucleic acids from biologics.

UltraNuclease, also referred to as broad-spectrum nuclease and non-specific endonuclease, can cleave and degrade various forms of DNA and RNA under multiple experimental conditions. It is widely used to remove nucleic acids from Biologics.

Product Application Scenarios

  1. Removal of Residual Nucleic Acids in Biologics: During the production of biologics, such as viruses, vaccines, and proteins,UltraNuclease can be added to degrade residual nucleic acids. This step not only increases the yield of viruses but also significantly reduces the content of residual nucleic acids, thereby ensuring the safety and efficacy of the drugs.
  2. Viscosity Reduction: By degrading nucleic acids, the viscosity of cell lysates is reduced, which facilitates filtration/ultrafiltration processes during the purification of cell-derived particles such as viruses, AAV vectors, and inclusion bodies.
  3. Improved Resolution and Recovery: In ELISA, two-dimensional electrophoresis, and immunoblotting analyses, it improves resolution and recovery.
  4. Prevention of Cell Aggregation: UltraNuclease can effectively prevent the aggregation of human peripheral blood mononuclear cells (PBMCs).

The effect of different reaction conditions on enzyme activity

    To explore the optimal operating conditions for UltraNuclease, we studied the effects of various buffering systems on enzyme activity.

    Comparative analysis of competing products (from A company)

    The performance of the UltraNucleaseis consistent with Company A, and it can achieve a replacement effect.

    The quality inspection standard conforms to the GMP production standard

    Detection items

    Quality standards

    Detection method

    Enzyme activity

    250-300 U/uL

    Universal substrate method

    Specific activity

    ≥1.5×10U/mg

    Universal substrate method

    Protein purity

    ≥99%

    SDS-PAGE

    Protease

    Meets the requirements

    Universal substrate method

    Bacterial endotoxin

    Meets the requirements

    Gel method

    Host protein

    Negative

    ELISA method

    Host DNA

    Negative

    qPCR method

    Sterile testing

    Sterile growth

    General Rule 1101

    Pathogen detection

    Negative (not detected)

    Refer to the internal method

    Mycoplasma testing

    Negative (not detected)

    Refer to the internal method

    Heavy metal testing

    Meets the requirements

    General Rule 0821

    To match the use of UltraNuclease, Yeasen has also independently developed a specific detection kit for the UltraNuclease, which can effectively detect residual enzyme and reduce risks. The Yeasen UltraNuclease Residual Detection Kit was developed in reference to the 2020 Chinese Pharmacopoeia <9101> Analytical Method Validation Guidelines, and strictly follows the key requirements for multi-parameter testing (linear range, specificity, accuracy, precision, sensitivity, accelerated stability, etc.).

    Residual UltraNuclease Detection

    In normal purification steps, UltraNuclease is easily removed as impurities. In order to detect the specific residual amount of omnipotent UltraNuclease, it is often used as a standard product in the industry and a matching ELISA kit is developed.

    The UCF.ME® UltraNuclease ELISA Kit independently developed by Yeasen adopts the principle of double-antibody sandwich enzyme-linked immunoassay (sandwich ELISA), which can accurately detect the residual amount of UltraNuclease in samples. The kit is stable in linearity, repeatability, recovery rate and specificity.

     

    Principle of Yeasen UltraNuclease ELISA Kit (Two-site immunoenzymetric assay)

    Product Features:

    Regulatory Compliance: Fully validated according to regulatory requirements, validation reports are available; Quality Assurance: All raw materials for the kit are independently developed, ensuring quality control;

    High Sensitivity: Quantitative limit as low as 23.5 pg/mL;

    High Precision: High intra-batch repeatability and low inter-batch variability;

    Strong Specificity: No cross-reactivity with other engineered cell proteins

    Performance parameters:

    Product parameters

    content

    Assay time

    3.5hours

    Assay principle

    Two-site immunoenzymetric assay

    Signal amplification

    Biotin-streptavidin system

    Detection wavelength

    450nm and 630nm

    Sensitivity

    23.5pg/mL

    Detection range

    0.047~3 ng/mL

    Detection object

    UltraNuclease

    Applicable instrument

    Molecular Devices: M and i Series

    Application

    Detection of residual UltraNuclease in purification processes of cell and gene therapy and recombinant adeno-associated virus vaccines

    Product data:

     Reagent kit standard curve:

    Actual sample recovery experiment:

    UltraNuclease was spiked in 5 μg/mL cell lysis solutions (HEK293, E.coli, CHO, and Vero) independently with a final spiked conc. of 1.5 ng/mL. The accuracy in different cell samples showed excellent than other vendor.

    Specificity demonstration

    Conclusion: The test kit has high specificity with no cross-reaction; pairwise comparisons, “*” Significant difference from Negative sample; “NS” No significant difference from Negative sample.

    Precision:

    Repeatability:

    Concentration ng/mL

    6 duplicated date in the same plate

    CV value

    3

    2.412

    2.390

    2.323

    2.367

    2.476

    2.291

    2.53%

    1.5

    1.780

    1.883

    1.884

    1.908

    1.876

    1.883

    2.19%

    0.75

    1.123

    1.215

    1.166

    1.183

    1.140

    1.109

    3.12%

    0.375

    0.691

    0.706

    0.708

    0.722

    0.722

    0.679

    2.21%

    0.1875

    0.449

    0.462

    0.453

    0.460

    0.447

    0.433

    2.13%

    0.094

    0.306

    0.308

    0.304

    0.311

    0.295

    0.302

    1.65%

    0.047

    0.242

    0.241

    0.240

    0.241

    0.256

    0.240

    2.37%

    0

    0.183

    0.192

    0.189

    0.187

    0.189

    0.189

    1.43%

    Conclusion: 6 duplicated were tested at each concentration on the standard curve, and all the CV% were lower than 5%.

    Intermediate precision:

    Intermediate precision

    Sample concentration

    1.5 ng/mL

    0.375 ng/mL

    0.094 ng/mL

    Analyst 1

    1.388

    0.382

    0.079

    Analyst 2

    1.458

    0.363

    0.068

    Analyst 3

    1.318

    0.37

    0.063

    Average

    1.388

    0.372

    0.07

    SD

    0.070

    0.010

    0.008

    CV value

    5.04%

    2.59%

    11.69%

    3 analysts conduct the UCF.ME™ UltraNuclease ELISA experiment and the CV% were lower than 15%

    Product Information:

    Product Name

    Product number

    Product Specifications

    UCF.ME® UltraNuclease (Benzonase)

    20156ES25/50/60/

    25KU/50KU/100KU

    UCF.ME®UltraNuclease GMP-grade (Benzonase)

    20157ES25/50/60/80/90

    25KU/50KU/100KU/1MU/5MU

    UCF.ME® UltraNuclease ELISA Kit

    36701ES59

    96T

     

    Other related products

    Product Name

    Product number

    Product Specifications

    Salt Active UltraNuclease GMP-grade

    20159ES25/50/60/80/90

    25KU/50KU/100KU/1MU/5MU

    Salt Active UltraNuclease ELISA kit

    36703ES96

    96T