A two-step strategy to visually identify molecularly imprinted polymers for tagged proteins

Alexander Brandis, Eran Partouche, Tamar Yechezkel, Yoseph Salitra, Vladimir Shkoulev, Avigdor Scherz, Flavio Grynszpan

Research output: Contribution to journalArticlepeer-review

Abstract

A practical and relatively simple method to identify molecularly imprinted polymers capable of binding proteins via the molecular tagging (epitope-like) approach has been developed. In our two-step method, we first challenge a previously obtained anti-tag molecularly imprinted polymer with a small molecule including the said tag of choice (a biotin derivative as shown here or other) connected to a linker bound to a second biotin moiety. An avidin molecule partially decorated with fluorescent labels is then allowed to bind the available biotin derivative associated with the polymer matrix. At the end of this simple process, and after washing off all the low-affinity binding molecules from the polymer matrix, only suitable molecularly imprinted polymers binding avidin through its previously acquired small molecule tag (or epitope-like probe, in a general case) will remain fluorescent. For confirmation, we tested the selective performance of the anti-biotin molecularly imprinted polymer binding it to biotinylated alkaline phosphatase. Residual chemical activity of the enzyme on the molecularly imprinted polymer solid support was observed. In all cases, the corresponding nonimprinted polymer controls were inactive.

Original languageEnglish
Pages (from-to)3358-3367
Number of pages10
JournalJournal of Separation Science
Volume40
Issue number16
DOIs
StatePublished - Aug 2017

Keywords

  • binding
  • biotin
  • molecularly imprinted polymers
  • protein
  • tag

All Science Journal Classification (ASJC) codes

  • Analytical Chemistry
  • Filtration and Separation

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