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Microarrays are high-throughput and relatively inexpensive assays that can be automated to analyze large quantities of data at a time. They are used in genome-wide studies to compare gene or protein expression under two varied conditions, such as healthy and diseased states. Microarrays consist of glass or silica slides on which probe molecules are covalently attached through surface functionalization. Most commonly, the slides are prepared through the chemisorption of silanes to silica...
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Polymer Microarrays for High Throughput Discovery of Biomaterials
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Synthesis of Clickable Coating Polymers by Postpolymerization Modification: Applications in Microarray Technology.

Laura Sola1, Francesco Damin1, Paola Gagni1

  • 1Istituto di Chimica del Riconoscimento Molecolare, CNR , Via Mario Bianco 9, 20131 Milano, Italy.

Langmuir : the ACS Journal of Surfaces and Colloids
|September 16, 2016
PubMed
Summary
This summary is machine-generated.

This study introduces a new polymer modification technique using click chemistry to create stable surfaces for biomolecule attachment, overcoming limitations of previous methods for microarray applications.

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Area of Science:

  • Polymer Chemistry
  • Surface Science
  • Bioconjugation

Background:

  • N,N-dimethylacrylamide (DMA), 3-(trimethoxysilyl)propyl methacrylate (MAPS), and N-acryloyloxysuccinimide (NAS) copolymers are used for microarray surfaces.
  • The N-hydroxysuccinimide (NHS) ester in NAS is prone to hydrolysis, limiting its application in certain biomolecule immobilization scenarios.
  • Limitations include challenges in microchannel derivatization and micro-/nanoparticle functionalization due to NHS ester instability.

Purpose of the Study:

  • To develop a novel polymer coating with enhanced stability and functionality for biomolecule immobilization.
  • To overcome the limitations associated with the hydrolysis of NHS esters in existing polymer coatings.
  • To enable efficient bioprobe attachment using click chemistry reactions for advanced microarray applications.

Main Methods:

  • Postpolymerization modification (PPM) of a parent copolymer (copoly(DMA-NAS-MAPS)) was performed.
  • The active ester groups were reacted with a bifunctional amine to introduce new functionalities.
  • Click chemistry reactions, specifically azide/alkyne or thiol/maleimide couplings, were utilized for bioprobe immobilization.

Main Results:

  • A new family of polymers was synthesized via PPM, featuring stable click functionalities.
  • The modified polymers exhibit improved coating stability and enable direct attachment of biomolecules from solution.
  • Quantitative yields and pH insensitivity were observed for the click chemistry reactions.
  • The click surfaces were successfully applied in solid-phase PCR for KRAS mutation genotyping.

Conclusions:

  • Postpolymerization modification offers a robust strategy to enhance polymer coating stability and introduce versatile functionalities.
  • Click chemistry provides an efficient and reliable method for bioprobe immobilization, overcoming limitations of traditional NHS ester chemistry.
  • The developed click-functionalized surfaces demonstrate significant potential for various microarray applications, including diagnostics and high-throughput screening.