Validation Procedure for Multiplex Antibiotic Immunoassays Using Flow-Based Chemiluminescence Microarrays.

Area of Science:

• Analytical Chemistry
• Biotechnology

Background:

• Small molecules, such as antibiotics, are often detected using indirect competitive immunoassays.
• Multiplex quantification of these molecules can be achieved with chemiluminescence microarray immunoassays.
• Traditional confirmatory methods like HPLC-MS are effective but time-consuming and costly.

Purpose of the Study:

• To validate multiplex immunoassays as confirmatory methods for detecting small molecules in food samples.
• To present a validation procedure for flow-through chemiluminescence microarrays according to EU guidelines.
• To demonstrate the simultaneous detection of 13 antibiotics in milk using a small molecule microarray.

Main Methods:

• Development and validation of a multiplex immunoassay using flow-through chemiluminescence microarrays.
• Application of the assay for simultaneous detection of 13 antibiotics in milk samples.
• Implementation of an internal calibration using a regenerable microarray chip.

Main Results:

• Successful validation of the multiplex immunoassay as a confirmatory method.
• Demonstration of simultaneous detection of 13 antibiotics in milk with high precision.
• Achieved low cost per analyte per sample, significantly less than HPLC-MS.

Conclusions:

• Multiplex immunoassays, specifically chemiluminescence microarrays, can be accepted as valid confirmatory methods.
• The developed method offers advantages of speed, automation, and minimal sample pretreatment.
• The regenerable microarray chip with internal calibration ensures precise analytical results at a reduced cost.