Asunto(s)
Artritis/inmunología , Antígeno B7-1/metabolismo , Células Dendríticas/inmunología , Antígenos CD/metabolismo , Artritis/patología , Antígeno B7-1/genética , Antígeno B7-2 , Separación Celular , Células Dendríticas/patología , Humanos , Técnicas In Vitro , Activación de Linfocitos , Prueba de Cultivo Mixto de Linfocitos , Glicoproteínas de Membrana/metabolismo , Fenotipo , ARN Mensajero/genética , ARN Mensajero/metabolismo , Líquido Sinovial/citología , Linfocitos T/inmunologíaRESUMEN
Methodologies for establishing standard curves on automated immunoassay systems are dependent on the computational capability of the instrument, its throughput, and, perhaps, the technical experience of the user. Here, factors constraining this approach are discussed in general and the procedures used for the Serono SR1 analyzer are considered in detail. The SR1 is a moderate-throughput, fully automated immunoassay system capable of performing tests for (currently) 18 analytes. The associated user-generated standard curves are utilized not only for interpolation of results but also for evaluation of system performance. The choices of curve-fitting method, weighting of data, acceptance criteria, and outlier rejection are taken out of the hands of the user and are made a vital and integral part of the system. I justify this approach and demonstrate how a sensible mathematical approach must be tailored to the configuration of the instrument.
Asunto(s)
Autoanálisis/instrumentación , Inmunoensayo/instrumentación , Humanos , Inmunoensayo/normas , Inmunoensayo/estadística & datos numéricosRESUMEN
Optical immunosensors employing evanescent wave techniques have the potential to address the requirements of the 'alternative site' market; however, this potential has yet to be realised. The development of 'direct' sensors, such as those using surface plasmon resonance (SPR), has been hampered by problems of non-specific binding and poor sensitivity to small molecules. 'Indirect' sensors (for example, those employing a fluorescently labelled reagent) overcome many of the problems of direct sensors but require more sophisticated instrumentation because of the low light levels detected. In an attempt to combine the best features of the two techniques, an indirect SPR fluoroimmunoassay (SPRF) technique has been investigated. The surface field intensity enhancement produced by SPR is used to boost the emission from a fluorescently labelled immunoassay complex at a metal surface. The potential of the method is demonstrated by assaying for human Chorionic Gonadotrophin (hCG) in serum. Enhanced sensitivity over conventional total internal reflection fluorescence (TIRF) and SPR techniques was achieved.