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Related Concept Videos

¹H NMR: Complex Splitting01:13

¹H NMR: Complex Splitting

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Related Experiment Video

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Flow Cytometric Bead Sandwich Assay Based on a Split Aptamer.

Luyao Shen1,2, Tao Bing1,2, Xiangjun Liu1,2

  • 1Beijing National Laboratory for Molecular Sciences, Key Laboratory of Analytical Chemistry for Living Biosystems, CAS Research/Education Center for Excellence in Molecular Sciences, Institute of Chemistry, Chinese Academy of Sciences , Beijing 100190, China.

ACS Applied Materials & Interfaces
|December 26, 2017
PubMed
Summary
This summary is machine-generated.

Researchers designed a novel split aptamer for detecting soluble l-selectin. This split aptamer-based cytometric bead assay (SACBA) offers a sensitive and selective alternative to traditional antibody-based methods.

Keywords:
flow cytometric bead assayl-selectinprotein detectionsandwich assaysplit aptamer

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Area of Science:

  • Biotechnology
  • Molecular Biology
  • Assay Development

Background:

  • Split aptamers offer potential for developing novel biosensors.
  • Limited knowledge of aptamer binding structures hinders split aptamer generation.
  • Antibody-based assays are common but can be improved.

Purpose of the Study:

  • To design a new split aptamer for l-selectin detection.
  • To develop a split-aptamer-based cytometric bead assay (SACBA).
  • To demonstrate SACBA's utility in detecting soluble l-selectin in human serum.

Main Methods:

  • Determined target-binding and structure-stabilizing moieties of the l-selectin aptamer Sgc-3b using DMS footprinting and mutation assays.
  • Generated a split aptamer by separating the duplex strand in the structure-stabilizing moiety.
  • Optimized the split aptamer sequence to minimize non-specific binding.
  • Developed and validated a split-aptamer-based cytometric bead assay (SACBA).

Main Results:

  • Successfully designed a functional split aptamer targeting l-selectin.
  • Developed SACBA, demonstrating good sensitivity and selectivity for soluble l-selectin.
  • Validated SACBA for detecting spiked l-selectin in human serum samples.
  • The developed strategies for split aptamer generation and assay design are efficient and practical.

Conclusions:

  • The study presents a novel method for generating split aptamers by understanding parent aptamer structure.
  • A simple, efficient, and practical split-aptamer-based assay (SACBA) was developed for soluble l-selectin detection.
  • SACBA shows promise as a viable alternative to antibody-based assays in diagnostics and aptamer engineering.