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Multimodal Analytical Platform on a Multiplexed Surface Plasmon Resonance Imaging Chip for the Analysis of Extracellular Vesicle Subsets
Published on: March 17, 2023
Sequential analysis of multiple analytes using a surface plasmon resonance (SPR) biosensor
J W Chung1, R Bernhardt, J C Pyun
1Korea Institute for Science and Technology Europe (KIST Europe fGmbH), Stuhlsatzenhausweg 97, 66123 Saarbruecken, Germany.
Journal of Immunological Methods
|April 4, 2006
Summary
A novel sequential analysis method using a surface plasmon resonance (SPR) biosensor allows for the simultaneous detection of two analytes in a single sample. This method demonstrates high accuracy and no cross-reactivity, proving effective for analyzing biomarkers like human chorionic gonadotropin (hCG).
Area of Science:
- Analytical Chemistry
- Biochemistry
- Biosensing Technology
Background:
- Multiplexed analysis of biological samples is crucial for accurate disease diagnosis.
- Existing biosensing methods can face challenges with cross-reactivity and sample complexity.
- Surface Plasmon Resonance (SPR) biosensors offer high sensitivity but require optimized methods for analyzing multiple analytes.
Purpose of the Study:
- To develop and validate a sequential analysis method for detecting two analytes simultaneously using a single SPR biosensor.
- To establish two distinct detection models to accommodate different analyte compositions and labeling requirements.
- To optimize the immunoaffinity capacity for enhanced analyte binding and sensor response.
Main Methods:
- Development of a sequential analysis protocol for a single SPR sensing region.
- Design of two detection models: one with label-free and labeled detection, the other with dual labeled detection.
- Preparation of standard curves for each model and sequential analysis of anti-bovine serum albumin (anti-BSA) antibodies and horseradish peroxidase (HRP).
- Optimization of analyte binding capacity by adjusting molecular recognition element concentration ratios.
Main Results:
- The sequential analysis method achieved errors of less than 6% for both models, indicating high accuracy.
- No significant cross-reactivity was observed between participating antigens and antibodies in either model.
- Optimization of immunoaffinity (IA) capacity using sensor response (R(max)) was successfully demonstrated.
- Feasibility was confirmed by detecting human chorionic gonadotropin (hCG) and human albumin (hA) in human urine samples using Model 2.
Conclusions:
- The developed sequential SPR biosensor method provides a reliable and accurate approach for analyzing multiple analytes in a single sample.
- This method minimizes cross-reactivity and allows for optimization of binding capacity, enhancing analytical performance.
- The successful detection of pregnancy-related biomarkers (hCG and hA) in urine highlights its clinical diagnostic potential.
