Future potential assay methods

Information gained from assays and future potential The application of esterases assays have provided the first direct evidence that cell lysis (as distinguished from loss terms such as grazing and sedimentation) may be an important factor in marine systems (see Kirchman, 1999). While the meaning of cell lysis, its incidence and ecological importance remain unclear (see Franklin et al, 2006), and there are concerns about the assay, we currently lack objective, independent methods with which to measure lysis (cf, Agusti and Duarte, 2002). In future, it may be possible to identify particular esterases that are truly found only within cells and choose substrates that are more specific for them. 

Information gained from assays and future potential So far, few field measurements have been made of caspase-like activities (Berman-Frank et al., 2004 Vardi et al., 1999), but the assay methods appear to be sensitive enough to allow use in natural communities. As work using cultures proceeds, and our understanding of cell death processes improves, assays of capase-like activity may offer an important means to distinguish different forms of cell mortality. Aside from bulk in vitro assays, the availability of ceU-permeable substrates, coupled with flow cytometry will provide improved resolution and specificity (e.g. Bidle and Bender, 2008). 

As mentioned earlier, the immunoassay can be done with a number of procedural modifications and in this instance one must substitute the isotope with another molecule (fluorescent dye, magnetic particle, enzyme) which can be measured and therefore serve as the source of tracer. For our initial studies we have chosen to use the enzyme immunoassay (EIA) system. At the present time the EIA is still in its infancy and although a number of successful EIA s have been developed the method cannot be considered a panacea (34). The future of this assay appears to be very bright and exciting, and there is considerable interest in the application of the EIA to problems in both microbiology and clinical medicine (34). Many of the procedures and protocols are derived from RIA procedures and the EIA, like the RIA, has the potential to be performed in a multitude of procedural variations but, for the purpose of this manuscript we will describe only the system we have chosen for our use. 

Abstract This chapter reviews the basic theory and applications of piezoelectric immunosensors. The immunosensor assay formats most often used are introduced as well as a brief explanation of the typical methods of measurement. Immobilisation is discussed, the importance of each characteristic, the basic techniques employed and a comparison of their performance as investigated by many researchers. The main historical developments of piezoelectric sensors and how these have led to early piezoelectric immunosensors are reviewed. Immunosensor applications and a comparison of sensor performance, for various analytes are summarised. The potential future of this field is also discussed. 

The recently elaborated multivitamin methods for B vitamins assay using LC with MS/MS detection, micellar LC, LC-DAD/MS, LC/ESI-MS or LC-IDMS systems offer better sensitivity and selectivity, and can be potentially used for thiamine determination in fortified foods and infant formulas. These methods have potential to improve performance in the future. 

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