A NOVEL METHOD OF ISOLATION OF TLR4 FROM CELL LYSATES OF MONOCULAR CELLS

摘要

A novel method of isolation of TLR4 from cell lysates of mononuclear cells comprising: collecting bovine adult filarial parasites (Setaria digitata); preparing aqueous extracts of setaria digitata (FAg) to produce affinity purified filarial glycoprotein (AgW); isolating human PBMCs from heparinised venous blood samples; subjecting the isolated human PBMCs to the step of incubation to produce PBMC lysates; coupling the said AgW with CNBR activated sepharose; loading the AgW coupled with sepharose and PBMC lysates to an affinity purification column to isolate the receptor recognizing AgW and detecting the reactivity of anti-receptor antibodies and anti-human TLR4 antibodies to the affinity purified receptor using peroxidase conjugated anti-rabbit IgG (in solidphase assay).