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Meso scale discovery and luminex comparative analysis of calbindin D28K
Samer Sourial1, Maritha Marcusson-Ståhl, Karin Cederbrant
1Molecular Toxicology, Safety Assessment, AstraZeneca R&D Södertälje, 151 85 Södertälje, Sweden. s.sourial@student.liverpool.ac.uk
Abstract:
The sensitivity of different renal regions to xenobiotics requires the development of a multiplex immunoassay for the simultaneous analysis of kidney biomarkers. Calbindin D28K is a distal tubule-specific protein that can be detected in urine under pathological conditions. In this study, a pair of anti-calbindin D28K antibodies was used in an immunoassay for the detection of calbindin D28K expression in rat and human kidney and urine. Comparative analysis of the immunoassay was performed on the Meso Scale Development (MSD) and Luminex platforms. Analysis on both platforms detected calbindin D28K concentrations between 100 ng/mL and 100 pg/mL. Luminex detected 10-fold the amount of calbindin D28K in samples analyzed as compared to MSD, whereas calbindin D28K level in rat and human urine was below detection limit in both platforms. The application of the immunoassays described herein may be useful in toxicological and pathological studies of distal tubular damage in rats and human.
Insights
This study developed a novel immunoassay to detect calbindin D28K, a kidney biomarker, in rat and human samples. The assay shows promise for studying distal tubular damage in toxicological research.
Area of Science:
- Nephrology
- Biomarker Discovery
- Toxicology
Background:
- Renal sensitivity to xenobiotics necessitates methods for simultaneous kidney biomarker analysis.
- Calbindin D28K is a distal tubule-specific protein indicative of kidney pathology.
- Multiplex immunoassays are crucial for comprehensive toxicological assessments.
Purpose of the Study:
- To develop and validate a multiplex immunoassay for detecting calbindin D28K expression.
- To compare the performance of Meso Scale Development (MSD) and Luminex platforms for calbindin D28K detection.
- To assess the utility of the immunoassay in rat and human kidney and urine samples.
Main Methods:
- Development of a specific immunoassay using anti-calbindin D28K antibodies.
- Comparative analysis of the immunoassay on MSD and Luminex platforms.
- Quantification of calbindin D28K in rat and human kidney tissue and urine samples.
Main Results:
- Both MSD and Luminex platforms detected calbindin D28K concentrations ranging from 100 ng/mL to 100 pg/mL.
- Luminex platform demonstrated 10-fold higher sensitivity for calbindin D28K detection compared to MSD.
- Calbindin D28K levels in rat and human urine samples were below the detection limit of both platforms.
Conclusions:
- The developed immunoassay effectively detects calbindin D28K in kidney samples.
- The Luminex platform offers superior sensitivity for calbindin D28K quantification.
- This immunoassay is a valuable tool for toxicological and pathological studies of distal tubular damage.
