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High affinity ligands from in vitro selection: complex targets

K N Morris1, K B Jensen, C M Julin

  • 1Department of Molecular, Cellular, and Developmental Biology, University of Colorado, Boulder, CO 80309, USA.

Proceedings of the National Academy of Sciences of the United States of America
|April 18, 1998
PubMed
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Systematic evolution of ligands by exponential enrichment (SELEX) can generate high-affinity DNA ligands for multiple targets simultaneously, even within complex biological mixtures like cell membranes. This method aids in dissecting intricate biological systems.

Area of Science:

  • Biochemistry
  • Molecular Biology
  • Genomics

Background:

  • Systematic evolution of ligands by exponential enrichment (SELEX) is an in vitro method for isolating high-affinity oligonucleotides against specific targets.
  • SELEX is typically performed using purified targets, limiting its application in complex biological systems.

Purpose of the Study:

  • To determine if SELEX can be applied to complex biological mixtures, using human red blood cell membranes as a model system.
  • To investigate the simultaneous generation of ligands for multiple targets within a complex mixture.

Main Methods:

  • Human red blood cell membranes were used as a model system for SELEX.
  • A secondary selection scheme, deconvolution-SELEX, was developed to isolate specific ligands from the mixture.

Related Experiment Videos

Main Results:

  • SELEX successfully generated ligands that bind to multiple targets within the complex membrane mixture.
  • The binding affinities of these ligands were comparable to those obtained against purified targets.
  • Deconvolution-SELEX enabled the rapid isolation of ligands targeting specific proteins of interest.

Conclusions:

  • SELEX is effective for generating high-affinity ligands against multiple targets in complex biological samples.
  • This methodology offers a potential approach for dissecting complex biological systems and identifying novel molecular interactions.