NGF and proNGF Reciprocal Interference in Immunoassays: Open Questions, Criticalities, and Ways Forward

摘要: The homeostasis between mature neurotrophin NGF and its precursor proNGF is thought to be crucial in physiology pathological states. Therefore, the measurement of relative amounts could serve as a footprint for identification disease states, diagnostic purposes. Since part proNGF, their selective with anti-NGF antibodies not straightforward. Currently, many immunoassays are available, while assays few validated by published information. question arises, whether commercially available able distinguish two forms. Also, since biological samples forms coexist, measurements one species affected presence other? We describe experiments addressing these questions. For first time, were measured together tested different immunoassays. Unexpectedly, found reciprocally interfere experimental outcome. interference also calls into widely used ELISA methods, applied where coexist. an immunoassay, needed. propose possible ways forward, towards development assay. In particular, use well αD11 antibody alphaLISA assay optimized incubation times would solution avoid mixed sample containing proNGF. Furthermore, we explored possibility measuring sample. But commercial kit detection does allow mouse brain tissues. SPR approach Our help understanding technical limits NGF/proNGF ratio samples, concrete solutions this problem.