888.3net新浦京游戏(国际)官网-Best Gaming Platform

Intelli Nuclease
Home > Technology > Reagent > Intelli Nuclease > Intelli Nuclease

Intelli Nuclease


Biological products produced in cells, such as viral vaccines and viral vectors, contain residual DNA from the host cell substrates. Theoretically, residual DNA could deliver activated oncogenes and potentially infectious viral genomes to patients and induce carcinogenic or infectious events. Regulatory agencies around the world have set regulatory limits for residual DNA to ensure patient safety. During the production process, manufacturers need to reduce the content of such impurities through specific methods to meet the regulatory requirements. In addition, nucleic acid substances produced during upstream cell culture or cell lysis can significantly increase the viscosity of the solution, affecting the efficiency of downstream process steps.

 

Intelli nuclease is an  omnipotent endonuclease expressed in recombinant E. coli, originally from Serratia marcescens. The enzyme is composed of two identical subunits (approximately 27.5KDa), is a non-specific nuclease that can completely and efficiently degrade any form of DNA and RNA (single-stranded, double-stranded, linear, circular, supercoiled) into oligonucleotide of 2-5 bases in length.

 

Applications

Ÿ   Remove DNA/RNA in protein purification process, to reduce viscosity, simplify clarification process, and effectively improve yields

Ÿ   Prepare samples for ELISA, chromatography, 2D electrophoresis, and blotting, improving resolution and recovery. Eliminate host cell DNA/RNA residues during vaccine and protein manufacturing, improving product safety.

Ÿ   Prevent cell clumping during cell culture, including PBMC, chimeric cell mixture preparation.

 

Reaction Conditions

Ÿ   Optimum reaction conditions: pH 6-10 (prefer pH 8-9), 1-2 mM Mg2+, 0-150 mM NaCI, 0-42°C (prefer 37 °C)

Ÿ   Enzyme activity will be significantly inhibited (reduction rate>50%) if monovalent cations >300 mM, PO43+>100 mM, (NH4)2SO4>100 mM

Ÿ   The enzyme is compatible with PMSF (1 mM), EDTA (1 mM), reducing agents (e.g. DTT) , detergents (e.g. Triton).

 

Manufacturing standard

Product Name

Nuclease, Serratia marcescens extracellular endonuclease

CAS No.

9025-65-4

Form

Active substance; monomer

Appearance

Colorless, no particles

Expression system

Non-pathogenic Escherichia Coli, BL21(DE3)

Source

Recombinant E. coli modified by nuclease gene

Formula

50 mM Tris-HCl, 100 mM NaCl, 5% glycerol, 2 mM Mg2+, pH8.0

Storage

-20 ℃, avoid repeated frozen-thaw

Endotoxin content

< 0.5 EU/mg

SEC-HPLC (%)

≥99%

SDS-PAGE (%)

≥99%

Molecular weight

27501.51 Da

Theoretical molecular weight

27500.81 Da

Enzyme activity

>250 U/μL


 Stability Data


Intelli Nuclease

Ÿ   0.4%(w/v) of TritonTM X-100 has no effect on enzyme activity, and 1.0%(w/v) of TritonTM X-100 hads inhibitory effect on enzyme activity.

Ÿ   PMSF, maleic acid and 2-mercaptoethanol have no effect on enzyme activity at 1 mM

Ÿ   EDTA-2Na inhibited about 60% of the enzyme activity at 1 mM concentration.

 

Reaction condition

Relative enzyme activity

Reaction condition

Relative enzyme activity

Reaction condition

Relative enzyme activity

Triton-0.4% (W/V)

>80%

PMSF-0mM

>80%

Control

100%

Triton-1.0% (W/V)

>20%

PMSF-1mM

>80%

EDTA-2Na-1mM

>60%

Maleic acid-1mM

>80%

PMSF-3mM

>80%

2-Mercaptoethanol

>80%

 

Ordering Information

Specifications

Part No.

500 KU/vial

DNRG0302500-GMP (in-stock)

1 MU/vial

DNRG0302001-GMP (in-stock)

5 MU/vial

DNRG0302005-GMP (in-stock

 





Contact Us
  • Tel.: +86 021 6434 0155
  • E-Mail: marketing@duoningbio.com
  • Address: Building 30, No. 1525 Minqiang Road, Songjiang District, Shanghai
Message

Copyright © Shanghai Duoning Biotechnology Co., Ltd. All Rights Reserved Sitemap | Technical Support: Reanod

WeChat
WeChat

Message-

Duoning Biotechnology

+86 021 6434 0155

XML 地图