The Enzyme-Linked Immunosorbent Assay (ELISA) is a highly sensitive and highly specific immunoassay technique widely used in in vitro diagnostics (IVD) and life science research.
Its core principle is based on the specific binding of an antibody to its target antigen. This binding event is detected using an enzyme conjugate (e.g., Horseradish Peroxidase/HRP or Alkaline Phosphatase/ALP), which catalyzes a chromogenic substrate (e.g., TMB or pNPP) to produce a measurable color signal. This allows for the qualitative or quantitative detection of target molecules like proteins, antibodies, or hormones.
Common ELISA formats include:
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Direct ELISA
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Indirect ELISA
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Sandwich ELISA
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Competitive ELISA
Among these, the Sandwich ELISA is considered the gold standard for detecting antigens (e.g., viral proteins, cytokines) due to its superior sensitivity and specificity.
The success of ELISA depends on several key components:
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High-quality coating antibodies/antigens
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Detection antibodies with high specificity and affinity
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Enzyme conjugates with high activity and stability
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Chromogenic substrates with high sensitivity and low background
As a robust and well-established platform, ELISA remains an indispensable core technology in clinical diagnostics, quality control, basic research, and drug development, valued for its relative operational simplicity, high throughput, and reliable results.
艳 李
