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Specific Detection of Infected Cells using Optical Drag-Tags in Laser Force Cytology

Use a generic, label-free approach to assay cultured cells in-process during viral vaccine production for extent of infection based on laser-force cytology (LFC).
Categories
Assays
Project status
100% Completed

Solution

Performance Period: 10/1/2020 to 1/31/2022

This proposal aims to use a generic, label-free approach to assay cultured cells in-process during viral vaccine production for extent of infection based on laser-force cytology (LFC). LFC has no requirement for the use of specific probes, giving a general test of viral infectivity, and has been applied to viral infectivity and in-process measurements in vaccine production. This particular application presents challenges; one is that the cells need to be isolated from microcarriers for LFC analysis and another is that any infection by adventitious agent (AA) needs to be discriminated from infection by the production viral carrier (VC).

We will address these by implementing a microfluidic device to isolate cells from microcarriers and by quantifying levels of AA and VC in spiked bioprocesses using high-affinity nucleic acid probes in a rapid, high-sensitivity form of electrophoresis (micelle-tagging electrophoresis, MTE). This information will be fed into a mathematical model of infection progress to better predict the concentration of infected and non-infected cells at time points and maximize the sensitivity of LFC. We will also demonstrate the effectiveness of LFC for non-viral biomanufacturing processes, and to establish MTE as a rapid and quantitative means to assay viral capsids in bioprocesses.

Impacts

General test for viral infectivity in cultured cells without requiring the use of specific probes

Reduce the amount of time required by an assay to detect virus in cultured cells thus lowering the cost of a contamination event and resulting in less drug supply lost

Cultured cells analyzed for infectivity in real time (hundreds of cells analyzed in less than 5 minutes)

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Project Lead

Carnegie Mellon University

Carnegie Mellon University

Participating Organizations

LumaCyte

LumaCyte

Merck Sharp & Dohme LLC

Merck Sharp & Dohme LLC

Rensselaer Polytechnic Institute

Rensselaer Polytechnic Institute