Preparative scale isolation of vaccine-grade plasmid DNA
NucleoBond RS – for high regulatory standards
Modular system for high-quality plasmid purification
- Endotoxin levels ≤ 0.01 EU/μg DNA
- Closed columns for pump or FPLC processing
- Scalable system for up to 200 mg of plasmid DNA
Large-scale plasmid purification for processes with high regulatory standards
In response to rapid advancements in cell and gene therapy as well as vaccine production, the need for top-notch plasmid DNA has increased significantly. Emerging applications, such as mRNA-based vaccines and gene therapy treatments, rely heavily on superior plasmid DNA as a fundamental starting material. One of the most challenging aspects of these applications is dealing with endotoxins – components of cell walls from gram negative bacteria. Here's where MACHEREY-NAGEL’s NucleoBond RS comes into play. By employing the NucleoBond RS workflow, you can achieve endotoxin levels ≤ 0.01 EU/μg DNA, ensuring compliance with stringent regulatory standards. NucleoBond RS columns are designed for large-scale isolation of vaccine-grade plasmid DNA from bacterial cultures, utilizing peristaltic pumps or FPLC systems meeting the highest quality requirements.
Plasmid DNA Specifications
| Parameter | Method | Criterion |
| Integrity | AGE / Fragment Analyzer | ≥ 90 % supercoiled form |
| Purity | UV spectrum | A260 / A280 1.8 - 2.0 A260 / A230 1.9 - 2.3 |
| Endotoxin levels | LAL / rFC | ≤ 0.01 EU/μg DNA |
| Host DNA | gPCR | ≤ 1 % |
| Host RNA | AGE Fluorometric Quantification | Not visible at 200 ng DNA ≤ 2 % |
| Host Protein | BCA | ≤ 1 % |
Application data – Large-scale plasmid DNA purification
The NucleoBond RS purification workflow enables a straightforward scalability of your plasmid purification
Plasmid DNA was purified from E.coli with the four different sizes of our NucleoBond RS columns. The figure shows the maximum amounts of plasmid DNA achieved per column size.
Plasmids isolated with NucleoBond RS columns show very high structural integrity
The supercoiled form of plasmid DNA is considered to be the desirable form for applications like gene therapy or vaccination. Regulatory standards specify that plasmids have to be present with more than 90 % supercoiled form. These data show that plasmids isolated with our NucleoBond RS columns meet these standards.
Vaccine-grade plasmid DNA with more than 90 % supercoiled form
Technical data from plasmid isolations using our NucleoBond RS columns show endotoxin levels ≤ 0.01 EU/μg DNA, A260/A280 purity ratios between 1.8 – 2.0, and percentages of supercoiled (ccc) form ≥ 90 %.
| NucleoBond RS 10 | NucleoBond RS 50 | NucleoBond RS 100 | NucleoBond RS 200 | |
| Endotoxins (EU/μg DNA) | 0.009 ± 0.004 | 0.007 ± 0.006 | 0.003 ± 0.002 | 0.003 ± 0.003 |
| A260 / A280 | 1.99 ± 0.04 | 1.97 ± 0.02 | 1.97 ± 0.04 | 1.98 ± 0.01 |
| Supercoiled form (ccc) in % | 92.3 ± 3.0 | 93.8 ± 2.1 | 93.0 ± 3.1 | 95.2 ± 2.1 |
NucleoBond RS product features at a glance
- Vaccine-grade plasmid DNA with endotoxin levels ≤ 0.01 EU/μg DNA
- Scalable system for up to 200 mg
- Compatible with peristaltic pumps or FPLC systems, such as ÄKTA™
- Flexible component configuration
- High-quality plasmids as starting material for applications, like lentivirus/AAV production or IVT-RNA (e.g. mRNA)
Ordering information
To ensure maximum flexibility for our customers, we offer all components needed for plasmid purification with our NucleoBond RS columns as standalone products. For more detailed information regarding equipment and quantities of individual components required per prep please check the user manual.
Columns
Columns
Inhalt 5 Preps
Inhalt 1 Preps
Inhalt 1 Preps
Inhalt 1 Preps
Buffers
Buffers
Accessories
Accessories
Inhalt 50 Preps
Inhalt 50 Preps
