An Automated Nanoparticle-Based Homogeneous Immunoassay for Determining Docetaxel Concentrations in Plasma

Abstract

This immunoassay is suitable for quantifying DTX in plasma with advantages of small sample size, no sample pretreatment, and the ability to be applied to a wide range of clinical analyzers. With the validation of this method, the application of DTX testing in clinical practice may gain wider acceptance for individualizing patient DTX dosing.

A Critical Review of Properties and Analytical Methods for the Determination of Docetaxel in Biological and Pharmaceutical Matrices

“…[65,66] Several analytical methods were developed for analysis of DTX, with the purpose of identification and quantification in biological samples (Table 1) and also in diverse matrices ( Table 2), such as delivery systems, that were presented in the previous section of this review. These methods include, mostly, immunoassays, [13] CE assays, [15] and chromatographic assays.…”

“…The immunoassay methods consist of a competitive reaction using selective anti-DTX monoclonal antibodies-coated nanoparticles followed by the agglutination quantification reaction measured by the absorbance at 660 nm. [13,14] The technique was compared with LC-MS/MS for determination of the DTX concentration in human plasma by Geng et al [14] The separation of DTX from plasma by LC-MS/MS utilized a C 18 Diamonsil column (150 mm £ 4.6 mm i.d. 5.0 mm particle size) with a mobile phase composed of 0.1% formic acid:acetonitrile (ACN) (40:60, v/v) and the mass spectrometry (MS) was performed in the positive ion multiple reaction monitoring (MRM) mode, with an ion transition of m/z 830.5>550.4, resulting in a retention time at 7.5 min.…”

“…5.0 mm particle size) with a mobile phase composed of 0.1% formic acid:acetonitrile (ACN) (40:60, v/v) and the mass spectrometry (MS) was performed in the positive ion multiple reaction monitoring (MRM) mode, with an ion transition of m/z 830.5>550.4, resulting in a retention time at 7.5 min. The immunoassay method was carried out according to the protocol presented by Cline et al [13] and using the DTX reagent kit (Saladax MyDocetaxel assay reagent kit). The absorbance was determined by the difference between the reaction at cycle 27 and cycle 13 without subtraction of a blank reagent.…”

“…Overall, those studies reported differences in the parameters of wavelength, mobile phase, stationary phase, and others, which will be addressed herein. [3,11,12] Moreover, other analytical methods reported for DTX quantification include immunoassay, [13,14] capillary electrophoresis (CE) [15] and ultraperformance liquid chromatography (UPLC), which presents many advantages, including the faster analytical run and the use of smaller volumes of solvents. [16][17][18] Another important issue regarding DTX analytical methods is the analytical matrix, which should not interfere in drug quantification.…”

Comparison of the docetaxel concentration in human plasma measured with liquid chromatography-tandem mass spectrometry (LC-MS/MS) and a nanoparticle immunoassay and clinical applications of that assay

“…These techniques are more specific and sensitive, but the expensive equipment and the complicated protocol make them ill-suited to routine measurement of docetaxel, and these drawbacks may hinder clinical TDM of docetaxel. A nanoparticle immunoassay based on turbidimetry and monoclonal antibodies that compete with docetaxel has been developed and preliminarily verified to be suitable for clinical TDM of docetaxel (16). Therefore, an alternate immunoassay that is simple, rapid, and costeffective would allow routine monitoring of docetaxel.…”