Adventitious Agents
When conducting tests to confirm the safety of a biological product, using multiple assays to ensure identification of contaminants is needed to guarantee patient safety. When it comes to adventitious agents, several assays used in combination ensure patient health and safety and are required by regulatory bodies including the European Medicines Agency (EMA) and the US FDA.
Two main types of assays used to identify adventitious agents are in vitro assays when mammalian cell lines are used for the production of biopharmaceutical products and in vivo assays that supplement in vitro assays to detect viruses that cannot be grown in cell culture.
Sartorius offers a wide range of in vitro and in vivo assays for detection of adventitious agents and supports manufacturers in selecting the optimal combination of tests for their safety strategy:
In vitro testing
- Detection of adventitious agents in three indicator cell lines
- Pre-studies to determine potential sample cytotoxicity
In vivo testing
- Viral safety evaluation of products derived from cell lines of human origin
- Assay for extraneous agents in viral vaccines for human use
- Assay for extraneous agents in cell substrates
- Custom in vivo assays for adventitious agents in viral vaccines
- Pre-studies to determine potential sample toxicity
Testing for adventitious agents varies by project. Our experts design a testing plan around your product development and manufacturing needs
Ready-to-use, quality-assured testing plans and prevalidated cGMP tests allow immediate access to specialized testing and faster results
Our cGMP-compliant assays align with European Pharmacopoeia (EP) and United States Pharmacopeia (USP) guidelines
Assay Types
Adventitious agent in vitro testing with a turnaround time of 14 or 28 days depending on production cell line.
Testing of:
- Master cell banks
- Master virus seed stock
- Unpurified bulk harvest
- End of production cell banks / control cells
Cell-based cytotoxicity assessment
Testing of:
- Unpurified Bulk harvest where cytotoxicity is expected
- Master virus seed stock
- Working virus seed stock
- Unpurified bulk harvest
- (Virus)-post neutralization by antiserum
- Non-infectious viruses that may still cause toxicity
Suite of in vivo assays for adventitious virus testing in accordance with the applicable EU and US regulations
Testing of:
- Master Cell Banks
- Master Virus Seed Stock
- Unpurified bulk harvest
- End of production cell banks / control cells
In vivo based toxicity assessment
Testing of:
- Unpurified Bulk harvest where cytotoxicity is expected.
- Master virus seed stock
- Working virus seed stock
- Unpurified bulk harvest
- (Virus)-post neutralization by antiserum.
- Non-infectious viruses that may still cause toxicity
In vitro assays for the detection of adventitious viruses are performed when mammalian cell lines are used to produce biopharmaceutical and biotechnology products. ICH guidelines require that samples from the master cell bank, master virus seed stock, end of production cell bank and unpurified bulk harvest be inoculated into cell cultures susceptible to human and animal viruses.
The detector cell lines identified in the guidelines include human and primate cells and cells that are representative of the species and tissue type of the production cell line.
Sartorius offers in vitro testing on a range of different detector cell lines and supports developers in selecting the tests and cell lines best suited for their product. Additional cell lines may be needed if there is a recognized risk of a particular virus which is not readily detected using the standard detector cells.
Sartorius’ in vitro assays typically have a 14-day turnaround time for non-primate production cell system and 28-day for human and primate production cell systems. The health of the cells is monitored by light microscopy and cell monolayers are tested at the end of the incubation period for hemadsorbing viruses.
Validated Detector Cell Lines
Cell Line | Species | Cell Line | Species |
|---|---|---|---|
Vero | Simian | A9 | Mouse |
MRC-5 | Human | MDBK | Bovine |
CHO-K1 | Hamster | MDCK | Canine |
324-K | Human | SP2/0 | Mouse hybridoma |
HeLa | Human | BHK21 | Hamster |
MA104 | Simian | HEK-293 | Human |
At critical points of the production process, manufacturers of biologics are required to perform in vivo assays for adventitious viruses. ICH guidelines require samples from the master cell bank, master virus seed stock and end of production cell bank to be inoculated into animals (adult and suckling mice and embryonated eggs) to reveal viruses that cannot grow in cell cultures.
The animal’s health is monitored and any abnormality is investigated. Other animal species, such as guinea pigs or rabbits may be tested if an identified risk might not be detected in the mice or eggs.
Sartorius offers complete in vivo testing services to our customers and provides consultancy and support including viral risk assessments to help manufacturers decide whether additional testing is required. In vivo testing normally has a turnaround time of 28 days.
