Methods for detecting contaminants in solutions containing glucose polymers

摘要

The invention relates to a method for detecting contaminants of glucose polymers, said contaminants being capable of acting in synergy with one another so as to trigger an inflammatory reaction, characterized in that it comprises an in vitro inflammatory response test using modified cell lines.

主权项

1. A method for detecting pro-inflammatory contaminants in a composition comprising glucose polymers, the method comprising:
contacting a plurality of cell lines with the composition comprising glucose polymers, each of the plurality of cell lines expressing one or more Toll-Like Receptors (TLRs) or Nucleotide-binding Oligomerization Domain-containing protein-like receptors (NOD), wherein the plurality of cell lines comprises:
a) a cell line expressing TLR2 and containing a reporter gene encoding a secreted form of alkaline phosphatase under the direct control of the TLR2 signaling pathway;b) a cell line expressing NOD2 and containing a reporter gene encoding a secreted form of alkaline phosphatase under the direct control of the NOD2 signaling pathway;c) a cell line expressing TLR4 and containing a reporter gene encoding a secreted form of alkaline phosphatase under the direct control of the TLR4 signaling pathway;d) a cell line expressing TLR2, TLR4 and NOD2 and containing a reporter gene encoding a secreted form of alkaline phosphatase under the direct control of the TLR2, TLR4 or NOD2 signaling pathway; ande) a negative control cell line; measuring activity of the secreted form of alkaline phosphatase; and identifying, based on the higher alkaline phosphatase activity in any of the cell lines a) to d) compared to the negative control, the presence, in the composition, of proinflammatory contaminants capable of activating one or more innate immunity receptors and triggering an inflammatory reaction.