Naslov
Real-Time PCR Bioassay for Measuring Human Interferon-beta Bioactivity

Autori
Miller, Larisa ; Khan, Gulraiz ; Lerner, Michaela

Vrsta, podvrsta i kategorija rada
Sažeci sa skupova, sažetak, znanstveni

Skup
International Society for Interferon and Cytokine Research (ISICR) annual meeting

Mjesto i datum
Oxford, Ujedinjeno Kraljevstvo, 16.09.2007. - 19.09.2007

Vrsta sudjelovanja
Poster

Vrsta recenzije
Međunarodna recenzija

Ključne riječi
qPCR; MxA; cell-based assay; bioactivity; interferon-beta

Sažetak
Introduction: Interferon Beta (IFN-beta) therapeutics is used as first line therapy for Multiple Sclerosis (MS). IFN-beta exerts its biological effects by binding to specific receptors on the surface of human cells. This binding initiates a complex cascade of intracellular events that leads to the expression of numerous interferon-induced gene products and markers. These include 2', 5'-oligoadenylate synthetase (OAS), Myxovirus resistance protein (MxA), anti-viral protection, beta2-microglobulin, and neopterin. Specific activity of IFN-beta is currently determined by an in vitro cytopathic effect bioassay (CPE) that employs lung carcinoma (A549) cells and encephalomyocarditis (EMC) virus. This assay is cumbersome, technically challenging, time consuming and expensive to outsource. Objective: Developing an innovative cell-based bioassay that is quick, robust, safe, and easy to perform. A bioassay that will be replacing currently employed CPE cellbased assays. Method: The new bioassay is based on the real time polymerase chain reaction (PCR) measurement of mRNA that results from IFN- induction of the OAS family or MxA genes in A549 cells. Conclusions: The IFN- PCR based assay is sensitive, simple, time efficient and has significant advantages over the established CPE assay. The assay is easily transferable to a commercial testing laboratory and could be used for product release and in a clinical setting.

Izvorni jezik
Engleski

Znanstvena područja
Farmacija, Biotehnologija

POVEZANOST RADA