Fluorescence polarization immunoassay FPIA

Fluorescence microscopy, 16 483 Fluorescence polarization (FP), 14 149-150 Fluorescence polarization immunoassay (FPIA), 12 97... 

Figure 2.3 Relationship between the concentration of teicoplanin in the serum of patients as determined using high-performance liquid chromatography (HPLC) versus fluorescence polarization immunoassay (FPIA) (r = 0.974). (From McCann et al., 2002.)...

Because of the requirement to use radioactive substances, RIAs are frequently being replaced by other immunologic assays, such as ELISA and fluorescence polarization immunoassays (FPIA) (Niemann et al. 1985). These have similar degrees of sensitivity. FPIAs are highly quantitative, as are RIAs, and ELISAs can be designed to be quantitative. 

Niemann, A., Oellerich, M., Schumann, G., Sybrecht, G.W. (1985). Determination of theophylline in saliva, using fluorescence polarization immunoassay (FPIA). J. Clin. Chem. Clin. Biochem., 23(11), 725-732. 

In studies performed by Arnold " and Sachs and Arnold, hair samples were treated with a solution of 1 M NaOH and boiled until the hair disintegrated. The hydrolyzed sample was neutralized with 1 M HCl. The same approach was employed by Kintz and Mangin. Hair digests were analyzed directly by the enzyme multiplied immunoassay technique (Emit , Syva Corporation, Palo Alto, CA) and fluorescence polarization immunoassay (FPIA, Abbott Laboratories, Abbott Park, IL). 

TDj = Abbott fluorescence polarization immunoassay (FPIA). RIA = radioimmunoassay. 

Measurement of Plasma Homocysteine by Fluorescence Polarization Immunoassay (FPIA) Methodology6... 

Martinus J, Klein L, Fino J, Shipchandler M, Wang P. A fluorescence polarization immunoassay (FPIA) for the quantitation of cortisol. Clin Chem 1985 31 956. 

Sample analysis requires specific techniques, such as fluorescence polarization immunoassay (FPIA) and high-performance liquid chromatography (HPLC). These techniques are not always available in the pharmacy and so sample analysis is not always done in Spanish hospital pharmacy services but in laboratories. However, it is a pharmacist who interprets results, makes recommendations, and follows up on patients, all as part of clinical activities to pursue better patient care. In all hospitals with such a pharmaceutical service, doctors and other members of the health team welcome the contribution of pharmacists, with their pharmacokinetic knowledge, to the rational use of drugs. 

Enzyme activities may also be measured in urine, cerebrospinal fluid, bone marrow cells or fluid, amniotic cells or fluid, red blood cells, leukocytes, and tissue cells. Cytochemical localization is possible in leukocytes and biopsy specimens (e.g., from liver and muscle). Under ideal conditions, both the concentration of the enzyme and its activity would be measured. Radioimmunoassay (RIA) and its alternative modes such as fluorescence immunoassay (FIA), fluorescence polarization immunoassay (FPIA), and chemiluminescence immunoassay (CLIA) (discussed later), can be used to measure enzyme concentration as well as other clinically important parameters. 

There have been several new developments in fluorescence polarization immunoassay (FPIA). Lackowicz and Terpetschnig reviewed the use of long-lifetime metal-ligand complexes in fluorescence polarization assays [154, 155]. New complexes with Re(l) and Ru(II) were described for the highly sensitive detection of high-molecular weight analytes by FPIA. Laser-induced fluorescence polarisation detection has been used by Yatscoff and coworkers in capillary electrophoresis detection (CE-LIFP) [156]. For the analyte cyclosporin picomolar detection limits were attained. 

The FPIA method can be potentially used either independently or as a complementary method to enzyme-linked immunosorbent assay (ELISA) and fluorescence polarization immunoassays (FPIA) technique. FCIA does not need polarization equipment and is not markedly influenced by light scattering effects. The FCIA technique can be expanded for analysis of enzymes and receptors including adaptation to fibro-optic techniques. 

Fig. 8.18 Principle of Fluorescence Polarization Immunoassay (FPIA). The rotational correlation time (0) is different for the free [(Re-L) -HSA and bound [(Re-L) -HSA]-anti-HSA-antibody fraction of the tracer.

These elegant techniques do not require separation of the bound and unbound fractions prior to measurement of response. Such assays offer advantages by simplifying and eliminating a step, but are more prone to matrix effects. An example of a homogeneous assay is fluorescence polarization immunoassay (FPIA, Figure 4.28). ... 

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