In Vivo Replacement

NGS as an ethical alternative to animals in biosafety testing and characterization.

What is in vivo replacement and why is it important?

The use of animal models in the characterization and biosafety testing of biologic medicines goes back several decades. Despite the well understood limitations, in vivo testing has remained a core part of most biologic cell line and viral vector characterization packages. This is because other more sensitive methods such as PCR or even ELISA have needed some knowledge of the contaminating species. Alternatively, agnostic cell culture-based methods will only detect a narrow range of adventitious pathogens dependent on cell type. Therefore, despite the limited ability for animal models to sensitively detect a broad range of virus types, in vivo testing has remained a complementary method – until now.

NGS approaches have long been used for the agnostic detection of unknown pathogens both clinically and in commercial biopharmacutical manufacture, especially in contamination investigation. However, the adoption of NGS as a replacement to animal models has been slow due to concerns about regulatory acceptance, as well as the sensitivity of NGS.

In collaboration with Charles River Laboratories, PathoQuest is the first GMP CRO to conclusively show the superiority of NGS over conventional in vivo biosafety testing and characterization methods. This, combined with the commitment from regulators to facilitate the removal of animal models from most submissions, means now is the ideal time to begin switching away from in vivo testing.

What we do

Modalities Tested:

  • mAbs and recombinant biologics
  • Viral vectors
  • Cell therapies
  • Vaccines
  • RNA based therapeutics
  • Cultured meat

Benefits of NGS

GMP validated – in vivo testing mostly GLP
Fast results – days vs weeks
Much more reliable and robust than animal models
No need to generate neutralizing antibodies
Specific identification of contamination
Meet corporate 3Rs objectives

Advantages of NGS over Animal Testing

WHY THIS APPROACH

ICH: Draft Guidline for Consultation Q5A (R2) viral safety evaluation of biotechnology products derived from cell lines of human or animal origin

FIND OUT MORE

Publication: Historical evaluation of the in vivo adventitious virus test and its potential for replacement with next generation sequencing (NGS)

FIND OUT MORE

Publication: Systematic evaluation of in vitro and in vivo adventitious virus assays for the detection of viral contamination of cell banks and biological products

FIND OUT MORE

PathoQuest in vivo Brochure

FIND OUT MORE

Our comparison data

COMPARISON OF LIMITS OF DETECTION FOR IN VIVO AND NGS ASSAYS FOR MODEL ADVENTITIOUS VIRAL AGENTS.

In vivo assays were performed using suckling mice, adult mice, and embryonated hens’ eggs. For ethical purposes, we chose not to perform all in vivo tests. For each virus category (defined in this publication), only the most sensitive reported ones were used.

Challenges solved

  • Difficulty in generating a neutralizing antibody
  • Incompatibility of test material with in vivo methods
  • Limited sample volume for in vivo characterization
  • Meeting corporate objectives to reduce animal use

“Due to the nature of our therapeutic it was not possible to generate a neutralizing antibody. The use of in vivo assays would therefore have been challenging as we anticipated adverse effects within the tests. PathoQuest was able to provide us an NGS based alternative characterization strategy which we were able to present to the regulators as a viable alternative to traditional in vivo methods.”

OTHER SERVICES

Adventitious Virus Testing

Detection of viral contamination within the manufacturing process and beyond.

READ MORE

nCats Integration Site Analysis

Characterisation of genetic modifications for clone selection, genetic stability and lot release

READ MORE

Identity Confirmation

Genetic characterization of viral and plasmid products for release.

READ MORE

Cell Line Characterization

Biosafety screening and stability testing of manufacturing cells.

READ MORE

HLA Genotyping

Characterizing and screening for novel and emerging cell therapies.

READ MORE

Raw Material Testing

Screening of high-risk inputs such as animal products and media.

READ MORE

Contact us

U.S.

+1 484 212 9360

466 Devon Park Dr
Wayne, PA 19087
United States

E: contact@pathoquest.com

Sign up for our latest news

France

+33 (0)1 70 82 17 90

Biopark -Bâtiment B,
11, rue Watt
75013 Paris, France

How can PathoQuest help?

Name(Required)
This field is for validation purposes and should be left unchanged.

U.S.

+1 484 212 9360

466 Devon Park Dr
Wayne, PA 19087
United States

France

+33 (0)1 70 82 17 90

Biopark -Bâtiment B,
11, rue Watt
75013 Paris, France

E: contact@pathoquest.com

How can PathoQuest help?

Name(Required)
This field is for validation purposes and should be left unchanged.

Sign up for our latest news