Rapid Adventitious Agent Testing for Fail-Fast Process Decision-Making

The aim of this project is to implement a nonPCR probe-based method for rapid adventitious agent testing as part of a fail-fast screening strategy of various process components to inform rapid process decisions thus reducing financial and supply risk
Categories
Drug substance
Assays
Project status
100% Completed

Industry Need

Current biomanufacturing includes a long turnaround time between sampling for adventitious agent testing, AAT, and availability of actionable assay results.

Solution

Rapid and frequent screening for likely AAs so that contaminated batches can be dealt with in the earliest stages. 

Outputs/Deliverables

  • Constructed a panel of MTE probes by a BLAST search to determine 10-15 nt sequences that appear in viral genomes and not in host
  • Developed a heating/organic modifier protocol that removed secondary structure of ssDNA/ssRNA targets to yield a single peak; stable γPNA binding allowed probes to remain bound
  • Demonstrated an MTE protocol that gives runtime less than 10 mins. with 10-100 pM detection level
  • Developed an isotachophoresis (ITP) + MTE protocol that gives detection of well under 1 fM viral genome in a 20-30 min run

Impacts

Rapid (< 1 hour) adventitious agent testing pathway for contaminated working cell bank cultures, raw materials, and bulk culture harvest lots.

Publications

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Project Lead

Carnegie Mellon University

Carnegie Mellon University

Participating Organizations

Bristol-Myers Squibb

Bristol-Myers Squibb