A monodispersed metal-complexing peptide-based polymer for mass cytometry enabling spectral applications

N Biotechnol. 2024 Jul 25:81:33-42. doi: 10.1016/j.nbt.2024.03.002. Epub 2024 Mar 16.

Abstract

We report the synthesis of a novel class of metal-complexing peptide-based polymers, which we name HyperMAPs (Hyper-loaded MetAl-complexed Polymers). The controlled solid-phase synthesis of HyperMAPs' scaffold peptide provides our polymer with a well-defined molecular structure that allows for an accurate on-design assembly of a wide variety of metals. The peptide-scaffold features a handle for direct conjugation to antibodies or any other biomolecules by means of a thiol-maleimide-click or aldehyde-oxime reaction, a fluorogenic moiety for biomolecule conjugation tracking, and a well-defined number of functional groups for direct incorporation of metal-chelator complexes. Since metal-chelator complexes are prepared in a separate reaction prior to incorporation to the peptide scaffold, polymers can be designed to contain specific ratios of metal isotopes, providing each polymer with a unique CyTOF spectral fingerprint. We demonstrate the complexing of 21 different metals using two different chelators and provide evidence of the application of HyperMAPs on a 13 parameter CyTOF panel and compare its performance to monoisotopic metal-conjugated antibodies.

Keywords: Coordination chemistry; Mass cytometry; Polymer; Rare earth metals; Spectral cytometry.

MeSH terms

  • Antibodies
  • Chelating Agents / chemistry
  • Coordination Complexes*
  • Maleimides*
  • Metals / chemistry
  • Peptides / chemistry
  • Polymers* / chemistry
  • Sulfhydryl Compounds / chemistry

Substances

  • Polymers
  • thiol-maleimide
  • Sulfhydryl Compounds
  • Peptides
  • Coordination Complexes
  • Metals
  • Chelating Agents
  • Antibodies
  • Maleimides