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Related Concept Videos

Enzyme-Linked Immunosorbent Assay01:33

Enzyme-Linked Immunosorbent Assay

In 1971, Peter Perlman and Eva Engvall developed an Enzyme-linked immunosorbent assay (ELISA or EIA). ELISA differs from western blot in that the assays are conducted in microtiter plates or in vivo rather than on an absorbent membrane.
There are many different types of ELISAs, but they all involve an antibody molecule whose constant region binds an enzyme, leaving the variable region free to bind its specific antigen.  Enzyme-substrate reaction allows the antigen to be visualized or quantified.

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Related Experiment Video

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Aptamer-Based Target Detection Facilitated by a 3-Stage G-Quadruplex Isothermal Exponential Amplification Reaction
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A self-assemble aptamer fragment/target complex based high-throughput colorimetric aptasensor using enzyme linked

Ji Nie1, Yan Deng, Qin-Pei Deng

  • 1Beijing National Laboratory for Molecular Sciences, Key Laboratory of Biochemistry and Molecular Engineering, College of Chemistry, Peking University, Beijing 100871, China.

Talanta
|April 20, 2013
PubMed
Summary

This study introduces a novel, high-throughput colorimetric aptasensor for detecting cocaine using a single aptamer sequence. This enzyme-linked aptamer assay (ELAA) offers a sensitive and specific method for molecular detection.

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Area of Science:

  • Biotechnology
  • Analytical Chemistry
  • Biosensors

Background:

  • Enzyme-linked aptamer assays (ELAA) utilize aptamers as recognition elements and enzymes for signal readout in aptasensor development.
  • Existing methods may require multiple aptamer sequences or complex procedures for target detection.

Purpose of the Study:

  • To develop a high-throughput, single-aptamer-based colorimetric aptasensor for cocaine detection using ELAA.
  • To demonstrate the versatility of this approach for detecting both small molecules and biomacromolecules.

Main Methods:

  • An anti-cocaine aptamer was fragmented, with one part immobilized on a 96-well plate and the other labeled with biotin.
  • Target binding facilitated the formation of aptamer fragment/target complexes, enabling detection via streptavidin-horseradish peroxidase (SA-HRP).

Main Results:

  • The aptasensor achieved a linear response for cocaine in the 5–200 μM range with a detection limit of 2.8 μM.
  • Specificity was validated, and the assay demonstrated successful application in real samples.
  • Thrombin detection confirmed the method's applicability to biomacromolecules.

Conclusions:

  • This single-aptamer ELAA provides a high-throughput, user-friendly, and specific colorimetric assay.
  • The approach offers a novel strategy for detecting diverse targets through aptamer fragment interactions.