Agmatine: Development of a clinical assay for therapeutic drug monitoring and neurological testing.

摘要

The aim of this work was to develop an Enzyme-Linked ImmunoSorbent Assay (ELISA) to be used for agmatine Therapeutic Drug Monitoring (TDM) when administrated to patients for neuropathic pain or in neurological research to understand the role of agmatine as a neurotransmitter in the central nervous system.; The ELISA was developed and improved in a series of six trials. In the first, we attempted to prove that an ELISA for agmatine was feasible for the range found in human plasma, as well as optimize the concentrations of reagents to work more accurately in that range. In the second, third, fourth and fifth trials we tried to improve the ELISA, making it shorter, easier, more user friendly and less expensive. Some of the intended improvements were achieved; some were not. When failure resulted, an explanation was given, as well as possible solutions for future developments, if the cost benefits justified overcoming the problems.; The last facet of our research was a clinical trial requiring the participation of patients and two medical doctors from MedCentral Health System to investigate agmatine levels in patients with stroke and epilepsy. Human samples were used to correlate the values obtained with the ELISA method and the previously developed GC-MS method. The clinical trial is still taking place, and it will require more time in order for us to recruit enough patients to obtain significant data, which will prove whether or not there is any relation between the value of agmatine and the incidence or progression of stroke or epilepsy. However, with the 69 patients' samples, we are able to confirm that we have a working ELISA technique for agmatine determination in human fluids, which is specific, sensitive, simple and reproducible.