Channel and substrate zone two-dimensional resolution for chemiluminescent multiplex immunoassay

Area of Science:

• Biomedical Engineering
• Analytical Chemistry
• Immunotechnology

Background:

• Multiplex immunoassays are crucial for simultaneous detection of multiple biomarkers.
• Existing methods can be complex and time-consuming.
• Development of efficient and automated multiplex assay systems is needed.

Purpose of the Study:

• To develop a novel two-dimensional resolution system for multiplex chemiluminescent immunoassay.
• To design and implement a multichannel detection device for simultaneous analysis of cancer biomarkers.
• To evaluate the performance and clinical applicability of the developed system.

Main Methods:

• A multichannel chemiluminescent detection device with a single photomultiplier was designed.
• Capture antibodies for CA 125, CA 153, CA 199, and CEA were immobilized in separate channels.
• A sequential sandwich immunoassay format with on-line incubation and CL substrate delivery was employed.

Main Results:

• The system successfully performed multiplex immunoassay for CA 125, CA 153, CA 199, and CEA with sensitive detection limits.
• Assay ranges achieved were 0.50-80 U/mL (CA 125), 2.0-100 U/mL (CA 153), 5.0-150 U/mL (CA 199), and 1.0-70 ng/mL (CEA).
• The entire assay process, including regeneration, was completed within 37 minutes with good reproducibility and agreement with clinical serum tests.

Conclusions:

• The proposed two-dimensional resolution system offers a simple, automated, and near-simultaneous approach for multianalyte immunoassay.
• This technique presents a new strategy for efficient biomarker detection in clinical diagnostics.
• The developed system demonstrates potential for high-throughput and cost-effective biomarker screening.