Detection of pyrogenicity on medical grade polymer materials using rabbit pyrogen, LAL and ELISA method

The objective of the study is to detect the pyrogenicity of five medical grade gelatinous polymer materials, intended for the manufacturing of capsule for pharmaceutical applications, by an indigenously developed ELISA, LAL and rabbit pyrogen assays. The ELISA methodology includes the incubation of the sample extract with blood from a healthy donor at 37 °C. Any pyrogen present in the extract induces the IL-1β which can be determined by ELISA. The rabbit pyrogen and LAL assays were performed as per standards. The result of the ELISA method indicated that all the materials extract induced high level of IL-1β as a marker for pyrogenicity. The rise in temperature of rabbit pyrogen was above 0.5 °C in all materials extract. LAL assay induced an endotoxin level above 0.5 EU. All the five polymer materials were found pyrogenic in all the assays. The ELISA method is very sensitive because the lowest limit of detection was 10 pg/ml endotoxin. Hence it can be concluded that the ELISA method will be an added advantage for the quality control release of a batch of medical products and improving the existing methodologies in the context of reduction and replacement in the use of animal models.