Product Class: Other

NEBExpress® Salt Active Nuclease, GMP Grade
Gray property icon for cloned at NEB Gray property icon for recombinant Gray property icon for engineered for performance

This product is available in a bulk or custom format. Contact us for more information.


Catalog #G8024

Product Introduction


NEBExpress® Salt Active Nuclease interacting with dsDNA fragment


  • Engineered nuclease that leverages high salt conditions for removal of nucleic acids (DNA and RNA) in biopharma and adeno-associated virus (AAV) manufacturing, and other bioprocessing workflows.
  • Manufactured to New England Biolabs’ GMP-grade* quality standards.
  • Formulated 10x more concentrated compared to the research-grade version (NEB #M0764)
  • 2X higher activity compared to other salt tolerant nucleases to increase productivity and reduce costs.
  • Degrades all DNA and RNA forms, including double-stranded and single-stranded DNA and RNA, linear and circular DNA and RNA, nucleosomal DNA, and DNA:RNA hybrids.
  • Recombinant enzyme, not glycosylated, formulated without detergents.

Product Information

Description

NEBExpress Salt Active Nuclease, GMP Grade is a recombinant, engineered broad-specificity nuclease, developed to leverage high salt conditions to degrade nucleic acid impurities in protein, adeno-associated virus (AAV), and biopharmaceutical manufacturing workflows. NEBExpress Salt Active Nuclease, GMP Grade is enzymatically identical to the research-grade product (NEB #M0764), degrading all forms of DNA and RNA at a broad range of salt concentrations, with optimal activity at 500 mM NaCl/KCl, and up to 2X more activity than other nucleases at high salt. Manufactured according to New England Biolabs’ GMP-grade* quality standards for stringent quality documentation and manufacturing needs, this GMP-grade enzyme is offered 10x more concentrated (at 1,000,000 Units/ml or 1 mg/ml) than the research-grade product (at 100,000 Units/ml).

Figure 1: NEBExpress Salt Active Nuclease outperforms two market-leading nucleases on 3 substrates at 500 mM NaCl

Bar graph comparing relative activity of NEBExpress® Salt Active Nuclease and nucleases from other suppliers

The percent activity of NEBExpress Salt Active Nuclease and two other nucleases (normalized by concentration in μg/ml) was determined by measuring the degradation of fluorescent oligonucleotide substrates (dsDNA, ssDNA, RNA) at room temperature in the optimal buffer for each nuclease adjusted to a final salt concentration of 500 mM NaCl.


Figure 2: NEBExpress Salt Active Nuclease, GMP Grade has exceptionally high physical purity and lot-to-lot consistency, as determined by microfluidic electrophoresis

Electropherograms of three lots

The purity percentage of NEBExpress Salt Active Nuclease, GMP Grade (lots normalized by concentration in μg/ml) is determined by microfluidic capillary electrophoresis. Samples are denatured and fluorescently stained, followed by separation based on electrophoretic mobility across the instrument's detection window. The instrument separates, sizes, and quantifies the fluorescently labeled bands detected against a sizing ladder to determine total purity. (i.e. desired protein products from impurities and degradants).

Chart 1: Attributes of NEBExpress Salt Active Nuclease, GMP Grade

Attributes Values
Protein Purity by Microfluidic Electrophoresis ≥ 95%
Identification by Microfluidic Electrophoresis Comparable to reference standard
Nominal Protein Concentration (A280) 1 mg/ml
Protease activity None detected
Endotoxin ≤ 5 EU/ml
Bioburden None Detected (Rapid) or
< 5 CFU/ml TAMC, (USP<61>)
< 5 CFU/ml TYMC, (USP<61>)
Unit Activity of NEBExpress Salt Active Nuclease 1,000,000 U/ml

With NEB’s stringent GMP-grade manufacturing and quality control practices, NEBExpress Salt Active Nuclease, GMP Grade is suitable for host cell nucleic acid removal in development and manufacturing workflows of viral vectors, protein therapeutics and vaccines. In adeno-associated virus (AAV) bioprocessing where high salt concentration is beneficial to minimize viral particle aggregation, this enzyme enables one step host cell lysis and removal of nucleic acid impurities.

Chart 2: Manufacturing of NEBExpress Salt Active Nuclease, GMP Grade through NEB’s GMP-grade processes

NEB GMP-Grade Quality & Manufacturing for NEBExpress Salt Active Nuclease, GMP Grade
Infrastructure
Infrastructure Icon
 Animal-free facility
Expanded qualification/validation requirements for facilities/utilities/environment/cleaning/process equipment/computer systems
ISO 8 Clean Rooms and ISO 5 Filling Hoods
Fermentation scales ranging from 1L up to 1000L and multiple largescale downstream purification, buffer formulation and final filling suites
Manufacturing processes
manufacturing icon
 Beta-lactam and cephalosporin-free process and facility
0.22 micron filtered final product
Certified Animal-free origin facility and process
Characterized master cell bank
Product attributes/testing
Product Attributes icon
 Enhanced panel of product contamination assays
Validated assays with quantitative results based on relevant principles of ICH Q2 Guidelines
TSE/BSE statement included on Certificate of Analysis for all GMP-grade batches
Compendial assays, including bioburden and endotoxin levels, where applicable
ICH Q1A/Q5C based product stability program
QA & Regulatory
QA Regulatory Icon
 ISO 9001:2015 and ISO 13485:2016 certified QMS
Full production batch records with defined critical quality attributes and critical process parameters
Regulatory supporting risk statements available, upon request, including but not limited to: melamine, residual antibiotic, mutagenic and elemental impurities, nitrosamine and residual solvents


Figure 3: NEBExpress Salt Active Nuclease outperforms market-leading nucleases across a range of NaCl concentrations

Line graph comparing relative activity of NEBExpress® Salt Active Nuclease and nucleases from other suppliers

NEBExpress Salt Active Nuclease is more active than other market-leading nucleases between 250 and 1,000 mM NaCl. The percent activity of each nuclease (normalized by concentration in μg/ml) was determined by measuring the degradation of a fluorescent dsDNA hairpin oligonucleotide at 25°C in the optimal buffer for each nuclease adjusted to a final salt concentration as indicated.

* “GMP Grade” and "GMP-grade" are branding terms NEB uses to describe products manufactured or finished at NEB’s Rowley facility. The Rowley facility was designed to manufacture products under more rigorous infrastructure and process controls to achieve more stringent product specifications and customer requirements. Products manufactured at NEB’s Rowley facility are manufactured in compliance with ISO 9001 and ISO 13485 quality management system standards. However, at this time, NEB does not manufacture or sell products known as Active Pharmaceutical Ingredients (APIs), nor does NEB manufacture its products in compliance with all of the Current Good Manufacturing Practice regulations.

 

Product Source

An E. coli strain that carries an engineered version of the endA gene from Aliivibrio salmonicida.
This product is related to the following categories:
Protein Tools,
DNA Modifying Enzymes & Cloning Technologies,
This product can be used in the following applications:
Protein Purification,
Protein Analysis Tools,
Protein Expression

Properties & Usage

Unit Definition

One unit is defined as the amount of enzyme required to release 32 pmol of FAM from FAM-BHQ1 labeled hairpin oligonucleotide in 1 minute at 25°C in a 50 µL reaction in 25 mM Tris-HCl, 500 mM NaCl, 5 mM MgCl2, 0.005% Tween 20 (pH 8.5 @ 25°C).

Storage Buffer

25 mM Tris-HCl
500 mM NaCl
5 mM MgCl2
50% Glycerol
pH 7.5 @ 25°C

Features

  • Manufactured according to NEB’s GMP-grade* quality standards and formulated at a high concentration of 1,000,000 U/ml (1 mg/ml).
  • GMP-grade endonuclease with broad nucleic acid specificity and optimally active at high salt concentration (500 mM NaCl or KCl)
  • Degrades double-stranded and single-stranded DNA and RNA, linear and circular DNA and RNA, nucleosomal DNA and both strands of DNA:RNA hybrids
  • Generates short oligos (as short as 5 nt) with 5´-phosphate and 3´-OH
  • Up to 2X more active than other comparable nucleases at 500 mM NaCl
  • Easy removal from a reaction by capture on cation exchange chromatography
  • Reactions are scalable and compatible with high-throughput experiments

 

This enzyme is ideal for:

  • Nucleic acid (DNA and RNA) removal at various steps of a bioprocessing workflow for proteins, viruses, or small molecules purification
  • Nuclease treatment during cell lysis in viral vector (e.g., AAV) manufacturing workflows
  • Nucleic acid (DNA and RNA) clearance in lysates to improve protein binding to affinity resins and reducing column back pressure
  • Reducing viscosity in cell lysates

Protocols, Manuals & Usage

Protocols

  1. General Guidance for NEBExpress® Salt Active Nuclease, GMP Grade Usage (NEB #G8024)

FAQs & Troubleshooting

FAQs

  1. I see that NEB offers two versions of NEBExpress® Salt Active Nuclease. What is the difference between NEBExpress Salt Active Nuclease (NEB #M0764) and NEBExpress Salt Active Nuclease, GMP Grade (NEB #G8024)?
  2. Can I order NEBExpress® Salt Active Nuclease, GMP Grade (NEB #G8024) at a larger volume or at a different concentration?
  3. What are the benefits of working in a high salt buffer?
  4. Does NEBExpress® Salt Active Nuclease have the same efficiency as Serratia marcescens nuclease?
  5. How does the unit definition for NEBExpress® Salt Active Nuclease compare to other similar salt-active nucleases?
  6. Can I use NEBExpress® Salt Active Nuclease in a low salt buffer?
  7. Is NEBExpress® Salt Active Nuclease compatible with affinity columns such as IMAC?
  8. Can I remove NEBExpress® Salt Active Nuclease from my sample?
  9. After use, how can I inactivate NEBExpress® Salt Active Nuclease?
  10. How are NEBExpress® Salt Active Nuclease and DNase I-XT (NEB #M0570) different?
  11. What is the molecular weight of NEBExpress® Salt Active Nuclease?
  12. Is NEBExpress® Salt Active Nuclease glycosylated?

Quality, Safety & Legal

Quality Assurance Statement

Quality Control tests are performed on each new lot of NEB product to meet the specifications designated for it. Specifications and individual lot data from the tests that are performed for this particular product can be found and downloaded on the Product Specification Sheet, Certificate of Analysis, data card or product manual. Further information regarding NEB product quality can be found here.
To request a certificate of analysis for this product please send request to info@neb.com

Legal and Disclaimers

Products and content are covered by one or more patents, trademarks and/or copyrights owned or controlled by New England Biolabs, Inc (NEB). The use of trademark symbols does not necessarily indicate that the name is trademarked in the country where it is being read; it indicates where the content was originally developed. The use of this product may require the buyer to obtain additional third-party intellectual property rights for certain applications. For more information, please email busdev@neb.com.

This product is intended for research purposes only. This product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.

New England Biolabs (NEB) is committed to practicing ethical science – we believe it is our job as researchers to ask the important questions that when answered will help preserve our quality of life and the world that we live in. However, this research should always be done in safe and ethical manner. Learn more.

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