Remicade TNF-Alpha Neutralisation Assays

The Remicade (infliximab) TNF-Alpha neutralization bioassay measures activity of the Fab binding affinity against TNF-Alpha, reporting the inhibition of TNF-Alpha mediated signal, commensurate to the concentration of Remicade (infliximab). BioOutsource are industry experts in the development, optimization and qualification/validation of bioassays and have applied this expertise to Remicade (infliximab) biosimilars and biobetters to offer a wide range of TNF-Alpha neutralization assays to our customers. We have provided services to support clone selection, characterization and biosimilarity activities with assays performed using our existing methodology, adapted to incorporate the customer’s reference standard of choice.

The primary mechanism of action of Remicade is the neutralization of circulating TNF-Alpha. TNF-Alpha itself is involved in an array of different signalling pathways and through binding to and neutralization of TNF-Alpha, Remicade has the capacity to neutralize the wide range of different biological activities that are mediated by TNF-Alpha. Historically, anti-TNF-Alpha therapeutic antibody products have used a murine cell line (L929 or WeHi) with a cell death endpoint as the primary potency bioassay. Increasingly, however Remicade biosimilar manufacturers are looking to perform more extensive characterization that is aligned with originator filings to study multiple different mechanisms of action of TNF-Alpha neutralization. BioOutsource currently offers six different versions of the Remicade TNF-Alpha neutralization assay:

  • Remicade TNF-Alpha Neutralisation Assay using L929 cells (Measurement of cell death/viability)
  • Remicade TNF-Alpha Neutralisation Assay using U937 cells (Measurement of cell death/viability)
  • Remicade TNF-Alpha Neutralisation Assay using U937 cells (Measurement of apoptosis)
  • Remicade TNF-Alpha Neutralisation Assay using HUVEC cells (Measurement of ELAM-1 Expression)
  • Remicade TNF-Alpha Neutralisation Assay using HUVEC cells (Measurement of ICAM-1 Expression)
  • Remicade TNF-Alpha Neutralisation Assay using HUVEC cells (Measurement of VCAM-1 Expression)

The principle of the Remicade (infliximab) neutralization bioassay is that the relevant cell line is prepared in cell plates. A dilution series of Remicade (infliximab) is then prepared and added to an optimized concentration of TNF-Alpha and incubated to allow neutralization to occur, prior to addition to the cell plate . The cell plate is then incubated for a further period of time before the assay is completed using the relevant detection methodology. Sample results can be reported as a per cent of inhibition of cell death or as a relative potency measurement against the reference standard.