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Multi-analyte validation in heterogeneous solution by ELISA
Thangavel Lakshmipriya1, Subash C B Gopinath2, Uda Hashim1
1Institute of Nano Electronic Engineering, Universiti Malaysia Perlis, 01000 Kangar, Perlis, Malaysia.
International Journal of Biological Macromolecules
|July 23, 2017
Summary
Enzyme Linked Immunosorbent Assay (ELISA) demonstrates effective multi-analyte detection in various solutions. This validated immunoassay shows promise for identifying multiple targets, including tuberculosis protein, Factor IXa, and cancer antigen, even in complex samples like human serum.
Area of Science:
- Biochemistry
- Immunology
- Analytical Chemistry
Background:
- Enzyme Linked Immunosorbent Assay (ELISA) is a widely used technique for detecting specific analytes in solution.
- Advancements in ELISA have expanded its applicability for a diverse range of targets.
- The need for efficient multi-analyte detection assays is growing in clinical diagnostics.
Purpose of the Study:
- To provide evidence for the suitability of ELISA in detecting multiple analytes simultaneously.
- To demonstrate the adaptability of ELISA for both homogeneous and heterogeneous solutions.
- To validate ELISA's performance with clinically relevant targets in complex biological matrices.
Main Methods:
- Adapted conventional Enzyme Linked Immunosorbent Assay (ELISA) protocols.
- Selected three distinct clinically relevant analytes: 16kDa protein (Mycobacterium tuberculosis), human blood clotting Factor IXa, and Squamous Cell Carcinoma (SCC) antigen.
- Performed assays in both homogeneous and heterogeneous solution formats, including human serum.
Main Results:
- Achieved sensitivities of 2 nM for 16kDa protein, 8 nM for Factor IXa, and 1 nM for SCC antigen in homogeneous and heterogeneous solutions.
- Demonstrated comparable sensitivities for multi-analyte detection in the presence of human serum.
- Confirmed the robustness of the adapted ELISA method for simultaneous detection.
Conclusions:
- Enzyme Linked Immunosorbent Assay (ELISA) is a viable and sensitive method for multi-analyte detection in heterogeneous solutions.
- The validated ELISA approach can be applied to a broader range of target analytes.
- This study supports ELISA's utility in complex biological samples, such as human serum, for simultaneous diagnostic purposes.
