Development of Standard Test Procedures for Quantifying Carry over from Fixed Pipetting Tips in Liquid-Handling Systems

Abstract

Robotic liquid-handling systems using fixed reusable pipetting tips are used not only in fully integrated in vitro clinical diagnostic analyzers, but also in open platform robotic liquid-handling systems. These are not confined to particular diagnostic assays, as customers adapt a large variety of assays on these platforms. One major problem with the use of fixed, reusable tips is the carry over (CO) of analyte from sample to sample. Despite widespread use of fixed tips in open platform systems, systematic studies on procedures to quantify CO for analytes other than nucleic acids are missing. In a consortium with three liquid-handling system suppliers and one coating specialist, we developed test procedures for the quantification of CO. The procedures were standardized and tested with the analytes fluorescein, immunoglobulin G, and hepatitis B surface antigen as model substances for small organic molecules, antibodies, and complex biomolecules. The test procedures allow the reproducible quantification of the CO with intra- and interassay precisions of less than 6% coefficient of variation. They were used to investigate the effect of different tip coatings on the CO of the three analytes. Fluoropolymers, inorganic-organic nanocomposites, sodium-silicate glass, titanium dioxide, and silicone resins, which are used in special applications, showed only small differences in CO. The CO test procedures can be easily transferred to different liquid-handling systems and used with different analytes.