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Viral Bioprocessing

APPLICATION overview, CHALLENGES AND SOLUTION

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For Detailed Info Including:
  • Product overview
  • Performance data & figures
  • Specifications
  • Documents
  • FAQs
  • Ordering Info
  • Protocols
  • Publications

Application Overview

Viral bioprocessing is pivotal for producing high-quality viral vectors used in gene therapy and vaccines. A key challenge has been salt’s detrimental effect on enzymatic function. Until now.

Our Salt Active Nucleases revolutionize the clearing of DNA because of their unique ability to clear DNA from viral vectors at your optimized salt conditions. 

We turn salt-induced challenges into a strength, streamlining the entire bioprocessing journey.

See enzymes for

For Detailed Info Including:
  • Product overview
  • Performance data & figures
  • Specifications
  • Documents
  • FAQs
  • Ordering Info
  • Protocols
  • Publications

Application Overview

Viral bioprocessing is pivotal for producing high-quality viral vectors used in gene therapy and vaccines. A key challenge has been salt’s detrimental effect on enzymatic function. Until now.

Our Salt Active Nucleases revolutionize the clearing of DNA because of their unique ability to clear DNA from viral vectors at your optimized salt conditions. 

We turn salt-induced challenges into a strength, streamlining the entire bioprocessing journey.

THE PROBLEM These ENZYMEs SOLVE

The Salt & Nuclease Paradox has long confounded the field of viral bioprocessing: As salt concentration increases, conditions for DNA clearance improve, but enzymes function decreases. This paradox becomes even more intricate when considering the interaction of chromatin, DNA binding, and the structural stability of enzymes in these conditions. 

In a nutshell, the result has led to compromised yields, impurities, and a more complex, costly bioprocessing journey.

In greater detail, the conditions for nuclease treatment can vary widely, influenced by vectors, serotypes, media, and downstream considerations. For instance, while fragile vectors like lentivirus may prefer DNA digestion under near-physiological conditions, robust vectors such as AAVs can benefit from higher salt concentrations. However, due to the high surface charge of some viral capsids, there's a tendency for them to associate with residual DNA, leading to aggregation and reduced viral titer. High salt buffers can mitigate this, making chromatin DNA more accessible for digestion. Yet, many commercially available endonucleases, often derived from Serratia marcescens, neither perform optimally in physiological conditions nor tolerate high salt concentrations. This mismatch in conditions inhibits nuclease activity, necessitating workflow adaptations or increased nuclease amounts.

THE PROBLEM These ENZYME SOLVES

The Salt & Nuclease Paradox has long confounded the field of viral bioprocessing: As salt concentration increases, conditions for DNA clearance improve, but enzymes function decreases. This paradox becomes even more intricate when considering the interaction of chromatin, DNA binding, and the structural stability of enzymes in these conditions. 

In a nutshell, the result has led to compromised yields, impurities, and a more complex, costly bioprocessing journey.

In greater detail, the conditions for nuclease treatment can vary widely, influenced by vectors, serotypes, media, and downstream considerations. For instance, while fragile vectors like lentivirus may prefer DNA digestion under near-physiological conditions, robust vectors such as AAVs can benefit from higher salt concentrations. However, due to the high surface charge of some viral capsids, there's a tendency for them to associate with residual DNA, leading to aggregation and reduced viral titer. High salt buffers can mitigate this, making chromatin DNA more accessible for digestion. Yet, many commercially available endonucleases, often derived from Serratia marcescens, neither perform optimally in physiological conditions nor tolerate high salt concentrations. This mismatch in conditions inhibits nuclease activity, necessitating workflow adaptations or increased nuclease amounts.

Relative nuclease activity for salt 0 – 500 mM
Fig 1. Relative nuclease activity for salt 0 – 500 mM

These enzymes not only maintain their functionality but excel in their respective salt environments. They ensure the most efficient DNA removal, even with the intricate dynamics of chromatin and DNA binding at play, paving the way for streamlined bioprocessing without the typical salt-induced challenges.

Relative nuclease activity for salt 0 – 500 mM

The Solution

ArcticZymes' salt-active nucleases, SAN HQ and M-SAN HQ, resolve the Salt & Nuclease Paradox. These enzymes not only maintain their functionality but excel in their respective salt environments. They ensure the most efficient DNA removal, even with the intricate dynamics of chromatin and DNA binding at play, paving the way for streamlined bioprocessing without the typical salt-induced challenges. 

SAN HQ and M-SAN HQ

Are especially developed to suit the high quality and regulatory requirements for use in bioprocessing workflows. The nucleases are manufactured according to requirements in ISO 13485. In addition relevant requirements from cGMP have been implemented.

SAN HQ GMP

The first salt active nuclease in GMP grade. SAN HQ GMP is biochemically identical to SAN HQ but produced under GMP conditions.

ELISA Kits for SAN HQ and M-SAN HQ 

ArcticZymes ELISA kits are used for the detection and quantification of SAN HQ or M-SAN HQ. The kits are developed as classical sandwich ELISAs and include monoclonal antibodies, specifically capturing trace amounts of SAN HQ or M-SAN HQ present in test samples.

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Relative nuclease activity for salt 0 – 500 mM
Fig 1.  Relative nuclease activity for salt 0 – 500 mM

These enzymes not only maintain their functionality but excel in their respective salt environments. They ensure the most efficient DNA removal, even with the intricate dynamics of chromatin and DNA binding at play, paving the way for streamlined bioprocessing without the typical salt-induced challenges.

Application Background

Cell and gene therapy are currently two of the most promising therapeutic areas, potentially allowing novel treatment of a variety of inherited and acquired diseases. In both therapies, engineered viral vectors are used to deliver and insert genetic material into cells to treat disease. Among the most promising tools in human gene therapy are viral vectors based on adenoviruses, adeno-associated viruses (AAVs) and lentiviruses. To drive clinical studies and commercialisation, the development of scalable, robust and high-yielding manufacturing methods for these vectors remains a key challenge for the industry.

At ArcticZymes Technologies, we have decades of experience as suppliers to the in vitro diagnostics industry. We prioritize both quality and cost-effectiveness for our customers. By providing top-tier products with full traceability and assisting clients in assay implementation, we believe we offer an unparalleled balance of value and quality in the market.

Given its optimal activity in high salt conditions, SAN High Quality improves downstream processes and reduces purification time without loss of vector yield and activity. SAN High Quality’s improved efficiency is serotype independent”.

- Lab Director of viral vector core at a major academic centre in the mid-Atlantic region

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