Solid Phase Reversible Immobilization magnetic beads has been used for nucleic acids purification because they are effective, simple, and fast. The carboxyl groups coated beads are paramagnetic particles that reversibly bind to nucleic acid. However, there is a drawback for magnetic beads that it can only purify DNA/RNA that are 100 base pairs or longer. tRNA and other DNA/RNA fragments shorter than 100 base pairs are not effectively recovered. We have developed Magnetic Beads (tRNA Purification) to overcome the hurdle.
Detail
Magnetic Beads (tRNA Purification)
Solid Phase Reversible Immobilization magnetic beads has been used for nucleic acids purification because they are effective, simple, and fast. The carboxyl groups coated beads are paramagnetic particles that reversibly bind to nucleic acid. However, there is a drawback for magnetic beads that it can only purify DNA/RNA that are 100 base pairs or longer. tRNA and other DNA/RNA fragments shorter than 100 base pairs are not effectively recovered. We have developed Magnetic Beads(tRNA Purification) to overcome the hurdle.
The Magnetic Beads(tRNA Purification) solves the tRNA and short DNA/RNA purification problem. The beads with our proprietary technology purify tRNA effectively by removing impurities and unwanted components such as dNTPs, detergents, salts, proteins, and other contaminants. The magnetic bead reagents can be used for oligo (>70 nt) applications.
Workflow without large RNA/DNA contamination
In the case of samples contaminated with RNA/DNA such as rRNA and DNA, our magnetic beads can effectively remove RNA/DNA that are 180 nt and larger. Purified tRNA are ideal for applications requiring high quality, as the fragments are free of impurities and contaminants.
Workflow with large RNA/DNA contamination
Comparison of tRNA and 70 nt oligos recovery. BioDynami beads (tRNA purification) successfully recovered DNA fragments both yeast tRNA and 70 nt oligos.
Recovery of tRNA and 70 nt oligos with BioDynami magnetic Beads (tRNA Purification). Yeast tRNA and 70 nt oligos were used as input. Input and recovered oligos were quantified with ssDNA Quantification kit (BioDynami Cat. # 40043) and RNA Quantification kit (BioDynami Cat. # 40044).
Depletion of larger RNAs. Left panel: depletion of 28S rRNA and 18S rRNA. Right panel: depletion of RNA of 180 nt and larger.
Features
Removal of unwanted components and other impurities
t-Boc-N-amido-Tri-(propargyl-PEG10-ethoxymethyl)-methane is reactive with azide-bearing compounds or biomolecules via copper catalyzed azide-alkyne Click Chemistry to yield a stable triazole linkage. The Boc group can be deprotected under mild acidic conditions to form the free amine. Reagent grade, for research use only.
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t-Boc-N-amido-Tri-(propargyl-PEG10-ethoxymethyl)-methane is reactive with azide-bearing compounds or biomolecules via copper catalyzed azide-alkyne Click Chemistry to yield a stable triazole linkage. The Boc group can be deprotected under mild acidic conditions to form the free amine. Reagent grade, for research use only.
Permagen’s 15 mL Centrifuge Tube Magnetic rack is designed for magnetic bead separations from up to six, 15 mL Centrifuge tubes
Accommodates any common 15 mL Centrifuge Tubes
Beads will be pulled to back wall allowing easy aspiration and tip tracking down the front wall of the tubes without disturbing bead pellet
Features include solid aluminum alloy design with hard coat finish for years of trouble-free use, rubber feet to help prevent slipping on work bench, less tippy than common plastic products, and fast separations using any magnetic beads
Bioprocessing with Salt Active Nucleases – High Salt Conditions
Product Info
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Product Info
Bioprocessing with Salt Active Nucleases – High Salt Conditions
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For SAN HQ, SAN HQ ELISA Kit, and now SAN HQ GMP
SAN HQ GMP is biochemically identical to SAN HQ but produced under GMP conditions.
Applications
Purification of biologics from residual nucleic acids in biopharma manufacturing
Purification of recombinant proteins and enzymes for research and diagnostic use
Removal of unwanted nucleic acids contamination in molecular biology reagents in challenging conditions
Reduction of viscosity in biological samples during production and automation
Vaccine manufacturing and viral vector preparation
DNA removal in high-salt lysates
SAN HQ – Peak performance at high salt conditions
Salt Active Nuclease High Quality (SAN HQ) is a Bioprocessing Grade nuclease developed as the most efficient solution for removal of both single and double stranded DNA and RNA at high salt conditions.
This nonspecific endonuclease has peak activity at salt concentrations between 400 – 700 mM (Fig. 1)
Non-enveloped viruses like Adenoviruses and Adeno-Associated Viruses (AAV’s) are inherently more robust with two distinct advantages: 1) They exhibit higher tolerance to additives like salt and detergents and 2) their production often involves the lysis of host cells, allowing for harvesting non-secreted vectors.
For Adeno-Associated Viruses (AAVs), which are often harvested from crude cell lysate, the high salt tolerance of SAN HQ is particularly beneficial. Salt is typically added to such lysates to reduce viral aggregation, facilitating more effective nuclease action to digest residual DNA.
SAN HQ’s is engineered for optimum activity in these high salt environments ensuring that you achieve unparalleled DNA removal without compromising the integrity of these robust viral vectors.
Key Benefits
Optimized Residual DNA Removal: Ensures efficient degradation of residual DNA in high-salt conditions, meeting stringent quality requirements for biologics and vaccines.
Boosted AAV Vector Purification: Enhances the purification process for adeno-associated viral vectors in high-salt conditions, improving quality and yield.
Streamlined Workflow: Eliminates the need for desalting stages, simplifying the bioprocessing protocol and saving time and resources.
Enable High-Throughput Processes: Facilitates scale-up and automation by working effectively in high-salt environments, increasing operational throughput.
Potential Surge in Virus Yield: Operates under conditions that may boost the titer yield of AAV production, potentially enhancing overall viral yield.
Economized Enzyme Usage: Reduces the need for excess enzyme and additional process adjustments, resulting in significant cost savings.
Minimized Risk of Process Disruptions: Offers reliable performance in various high-salt bioprocessing conditions, reducing the likelihood of disruptions due to enzyme inhibition.
Reliability: Provides consistent enzyme activity in challenging high-salt conditions, adding a layer of predictability and dependability to your operations.
Broader Applicability: Versatile enough to be used in a wide range of viral vector systems, expanding your research and production capabilities.
Enhanced Viral Stability: High-salt levels stabilize viral vectors, and SAN HQ operates effectively in these conditions, maintaining high yield and quality.
Host Cell Lysis: Facilitates efficient lysis of host cells in high-salt conditions, optimizing the harvest of both secreted and non-secreted viral vectors.
Key Features
High purity (≥ 98%)
No protease detected
Supplied with extended product documentation
Compatible with SAN HQ ELISA
The Challenge in Removing Host Cell Chromatin Impurities
In bioprocessing, the primary role of a nuclease is to efficiently digest and fragment host-cell DNA into sufficiently small pieces, facilitating its removal during downstream processing. While most nucleases can effectively degrade naked DNA into tiny fragments under optimal conditions—as demonstrated by M-SAN HQ and SAN HQ, which can digest dsDNA into fragments smaller than 6 nt—the reality in bioprocessing is more complex. (See fig. 5)
The DNA targeted for removal often exists as chromatin, embedded in a complex matrix containing remnants of the lysed host cell as well as large amounts of the therapeutic product.The product may or may not have an affinity for the chromatin you aim to remove.
High salt is often applied to mitigate issues like aggregation. The real challenge lies in a nuclease’s ability to efficiently fragment chromatin under these more complicated, high-salt, conditions—not merely degrading naked DNA under ideal circumstances.
SAN HQ ELISA kit is developed for the detection and quantification of SAN HQ and SAN HQ GMP. The kit is designed as a classical sandwich ELISA, with two monoclonal antibodies specific towards SAN HQ nuclease (fig 6).
Features
Sensitive: 0.4 – 25.6 ng/ml
Precise: RSD ≤ 15%
Accurate: 100% ± 15%
Stability: 12 months when stored between +2°C to +8°C
Document
For SAN HQ, SAN HQ ELISA Kit, and now SAN HQ GMP
SAN HQ GMP is biochemically identical to SAN HQ but produced under GMP conditions.
