Article

Understanding EndoCleave's role in viral vector, vaccine and biopharma manufacturing

EndoCleave: The "Technical Twin" endonuclease for early development and commercial scale 


EndoCleave is a high-purity (≥ 99%), animal-origin-free, recombinant endonuclease (produced in E. coli) designed to efficiently degrade all forms of DNA and RNA. Specifically optimized for viral Vector (AAV, lentivirus), vaccine and biopharma workflows, it effectively reduces lysate viscosity and removes host-cell nucleic acids.

Available in High Quality (HQ) and GMP Grade, EndoCleave acts as a "Technical Twin"—offering compositional equivalence to ensure a cost-effective and seamless transition from early process development to clinical manufacturing.  
 

We have compiled answers to the most frequently asked questions for you.
Please select below or just scroll down to find detailed information about EndoCleave:

General product information
Applications and strategy
Technical specification and conditions
Quality and regulatory 
Ordering and sampling

General product information

Where does EndoCleave originate and how is it produced?

EndoCleave is a recombinant endonuclease derived from Serratia marcescens. Unlike some market alternatives produced in bacillus species, EndoCleave is expressed in a proprietary Roche E. coli strain.

This production host is the industry standard and allows for animal-origin-free (AOF) and antibiotic-free (AB-free) production, ensuring a high safety profile with very low risk of protease contamination.

How does EndoCleave work?


EndoCleave works by cleaving the phosphodiester bonds in nucleic acids, effectively breaking them down into smaller oligonucleotides (2–5 bases) or mononucleotides. This process ensures the comprehensive removal of contaminating host-cell DNA and RNA, reducing lysate viscosity and facilitating clearer downstream purification.

 

What are the differences between EndoCleave High Quality (HQ) and EndoCleave GMP Grade?

EndoCleave High Quality is the "Technical Twin" to EndoCleave GMP Grade.

Differences:
EndoCleave GMP Grade is manufactured and tested to meet GMP guidelines (aligned with 21 CFR 820 & ISO 13485:2016), comes with extensive validation documentation and quality documentation (e.g., GMP Grade statement, impurities), and is intended for clinical and commercial manufacturing.

Similarities:
Both grades are produced using the identical host (E. coli), identical manufacturing process, and identical purity specifications (≥ 99%). The Certificate of Equivalence provides information confirming 1:1 compositional equivalence.

Applications and Strategy

What are typical applications for EndoCleave?

EndoCleave is optimized for use in Cell & Gene Therapy (CGT) and Biopharma workflows:

  • Early research: Supports discovery, optimization, and toxicity studies ("proof of concept"). The HQ grade enables the cost-effective screening of therapeutic candidates using a "clean", high-purity enzyme that de-risks the transition to animal models.
  • Process development: The HQ grade is ideal for "purification scouting" and optimization at pilot scales (e.g. 50L–200L).

  • Viral vector production: Essential for processing AAV, Lentivirus, and oncolytic viruses by eliminating host-cell nucleic acids and preventing cell clumping.

  • Viral vaccine production: Used to maximize throughput and yield in high-volume vaccine manufacturing (human and veterinary).

  • Protein purification: Efficiently removes nucleic acids to purify recombinant protein samples.
EndoCleave

Why should I use EndoCleave High Quality during early R&D instead of a generic "Lab Grade" enzyme?

Using EndoCleave HQ in early research serves as a "Bridge to the Clinic". Since EndoCleave HQ shares the same host and purity (≥ 99%) as the GMP Grade, data generated at the bench remains valid when you scale up. This mitigates the "technical drift" and costly process re-validation often required when switching from a generic low-purity R&D enzyme to a GMP Grade raw material.

Technical specifications & conditions

Viral Vector EndoCleave Roche CustomBiotech

What are the optimal conditions for using EndoCleave?

EndoCleave is versatile and effective across a broad range of conditions:

  • Temperature: Active between 4°C to 45°C (Optimal: 37°C).

  • pH range: Effective between pH 6.0 and 10.0 (Optimal: pH 7.5 - 8.5).

  • Cofactors: Requires the presence of divalent cations (Mg²⁺, 1–2 mM) for activity.

  • Inhibitors: Sensitive to chelating agents (e.g., EDTA) and high ionic strength.

How does EndoCleave’s purity compare to other market options?

Both EndoCleave HQ and GMP Grade offer a purity level of ≥ 99%. This exceptional quality provides a significant performance advantage over alternative industry options that typically offer purity levels above 90%.

By utilizing Roche’s high-purity HQ grade during early-stage research, you ensure that your process development is built on a material that accurately reflects the quality and consistency of the final GMP product. This alignment minimizes variables and streamlines the path from bench to production.

How can I detect potential residual EndoCleave during my process?

EndoCleave is engineered for full compatibility with standard commercially available ELISA kits designed to detect endonuclease from Serratia marcescens.

This design ensures that you can maintain your established analytical workflows without the need to modify existing quality control protocols or revalidate new detection methods.

Quality and regulatory

animal origin free

Is EndoCleave animal-origin-free (AOF)?



Yes. Both EndoCleave HQ and EndoCleave GMP Grade are fully animal-origin-free (AOF) and antibiotic-free (AB-free). This minimizes the risk of contamination (e.g., TSE/BSE) and potential allergic reactions, supporting regulatory safety requirements for use in both your human and veterinary applications.

regulatory documents

What supporting documents are available for regulatory needs?

Documentation packages vary by grade to suit the specific stage of development:

  • EndoCleave GMP Grade: Comprehensive package including Certificate of Analysis (CoA), Certificate of Origin, GMP Statement, Viral Safety Statement, Impurity Statements (Endotoxin, Bioburden, Heavy Metals), and support for regulatory submissions.

  • EndoCleave HQ: Includes Certificate of Analysis (CoA), Certificate of Origin, AOF/Antibiotic-Free Statements, and Safety Data Sheet (SDS). It does not include a GMP certificate but is produced under ISO 13485:2016 standards.

Both HQ and GMP Grade: Roche issues a Certificate of Equivalence.
This document confirms the compositional equivalence, which is essential for transitioning from early research and process development phases to clinical manufacturing.

Ordering and supply

How is EndoCleave supplied?

EndoCleave is available as a liquid solution (activity 250 – 350 kU/mL) in the following pack sizes:

 

  • 100 kU (0.4 mL): Available for HQ and GMP.
    Ideal for early development and small-scale research.
  • 500 kU (2.0 mL): Available for HQ and GMP.
    Suitable for mid-scale production and process optimization.
  • 5000 kU (20.0 mL): Available for GMP Grade only.
    Perfect for large-scale commercial manufacturing.

Can I request a sample for testing?

 Yes. Roche CustomBiotech offers free-of-charge test samples of EndoCleave HQ and GMP Grade (100 kU size) for preliminary technical evaluation and "head-to-head" comparisons.

Request your EndoCleave endonuclease sample here

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    Regulatory disclaimer, warranty limitations and legal notes are listed on respective product pages.