Propargyl-PEG3-bromide

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.

See Case Study on Efficient Fragmentation: Superior Chromatin Digestion Before Flow-Through Purification

SAN HQ ELISA Kit

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).

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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

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.

Description 

The ExcelRT™ One-Step RT-qPCR kit (TaqMan, ROX) is designed for reverse transcription and quantitative real-time analysis of a specific target RNA by one-step reaction. The ExcelRT™ One-Step RT-qPCR kit (TaqMan, ROX), consisting of One-Step RT Enzyme Mix and 2X One-Step Master Mix, is a convenient kit designed for highly efficient cDNA synthesis and highly specific real-time PCR in a single tube. The One-Step RT Enzyme Mix contains a thermostable ExcelRT™ Reverse Transcriptase and a RNAok™ RNase inhibitor. Consequently, One-Step RT Enzyme Mix can reverse transcribe RNA to cDNA at a wide temperature range from 42 to 60°C and active against RNase A, RNase B and RNase C. By containing specialized hot-start Taq DNA polymerase, which greatly reduce primer-dimer formation and can be activated within 2 minutes, the 2X One-Step Master Mix features high specificity and is suitable for fast cycle program. This master mix includes ROX reference dye for normalization of each RT-qPCR assay.

Features

• Reverse transcription at wide temperature range (42-60°C)
• High specificity 
• Suitable for fast cycle program
• With ROX reference dye

Storage

Aliquot to avoid multiple freeze-thaw cycles (stable within 30 freeze-thaw cycles)

Protect from light

-20°C for 12 months

The ExcelRT™ One-Step RT-qPCR kit (TaqMan, ROX) is designed for reverse transcription and quantitative real-time analysis of a specific target RNA by one-step reaction. The ExcelRT™ One-Step RT-qPCR kit (TaqMan, ROX), consisting of One-Step RT Enzyme Mix and 2X One-Step Master Mix, is a convenient kit designed for highly efficient cDNA synthesis and highly specific real-time PCR in a single tube. The One-Step RT Enzyme Mix contains a thermostable ExcelRT™ Reverse Transcriptase and a RNAok™ RNase inhibitor. Consequently, One-Step RT Enzyme Mix can reverse transcribe RNA to cDNA at a wide temperature range from 42 to 60°C and active against RNase A, RNase B and RNase C. By containing specialized hot-start Taq DNA polymerase, which greatly reduce primer-dimer formation and can be activated within 2 minutes, the 2X One-Step Master Mix features high specificity and is suitable for fast cycle program. This master mix includes ROX reference dye for normalization of each RT-qPCR assay.

The Opentrons® OT-2 is a high-precision, open-source liquid handler designed to be easy for scientists to use and flexible enough to automate many application workflows. This robot and its accompanying automation platform are used to automate hundreds of protocols and workflows in genomics, proteomics, cell-based assays, and drug discovery.

The OT-2 has a user-friendly, plug and play graphical interface to design protocols compatible with any ANSI/SBS compliant or automation friendly labware. The base model includes an Opentrons GEN2 Single-Channel Pipette; you can also use up to two pipettes at the same time. Additional pipettes can be purchased separately and are easily interchangeable by the user. Consumables and other labware are sold separately.

The Opentrons® OT-2 is a high-precision, open-source liquid handler designed to be easy for scientists to use and flexible enough to automate many application workflows. This robot and its accompanying automation platform are used to automate hundreds of protocols and workflows in genomics, proteomics, cell-based assays, and drug discovery.

The OT-2 has a user-friendly, plug and play graphical interface to design protocols compatible with any ANSI/SBS compliant or automation friendly labware. The base model includes an Opentrons GEN2 Single-Channel Pipette; you can also use up to two pipettes at the same time. Additional pipettes can be purchased separately and are easily interchangeable by the user. Consumables and other labware are sold separately.