Inflammatory response to pyrogens determined by a novel ELISA method using human whole blood

Abstract

Presence of pyrogens on implants, medical devices, drugs and biological materials compromise on the biosafety and poses a major health hazard in therapeutics. Detection of pyrogenic contamination has so far been done with either in vivo rabbit pyrogen assay or Limulus Amoebocyte Lysate (LAL) methods, each of which having their distinct advantages and disadvantages. An indigenously developed ELISA method quantifying the pro-inflammatory response triggered by pyrogens on human whole blood is demonstrated for its versatility to detect the pyrogenic response to gram-negative, gram-positive bacteria, chemical and biological pyrogens. The method was used to test and quantitate the pyrogen levels in polymeric biomaterials. Unlike the existing pyrogen test procedures, this assay is adapted to detect all pyrogens, besides yielding faster, sensitive and quantifiable data, thereby reduce/replace animal experimentation. The method also provided insight into the possible correlation between variable blood profile among individuals and their role in determining inflammatory response to different pyrogenic stimuli.