Highly bright avidin-based affinity probes carrying multiple lanthanide chelates

Laura Wirpsza, Shyamala Pillai, Mona Batish, Salvatore Marras, Lev Krasnoperov, Arkady Mustaev

Research output: Contribution to journalArticlepeer-review

6 Scopus citations

Abstract

Lanthanide ion luminescence has a long lifetime enabling highly sensitive detection in time-gated mode. The sensitivity can be further increased by using multiple luminescent labels attached to a carrier molecule, which can be conjugated to an object of interest. We found that up to 30 lanthanide chelates can be attached to avidin creating highly bright constructs. These constructs with Eu3+ chelates display synergistic effect that enhance the brightness of heavily modified samples, while the opposite effect was observed for Tb3+ chelates thereby significantly reducing their light emission. This undesirable quenching of Tb3+ luminophores was completely suppressed by the introduction of an aromatic spacer between the chelate and the protein attachment site. The estimated detection limit for the conjugates is in the 10-14-10-15 M range. We demonstrated a high sensitivity for the new probes by using them to label living cells of bacterial and mammalian origin.

Original languageEnglish (US)
Pages (from-to)22-29
Number of pages8
JournalJournal of Photochemistry and Photobiology B: Biology
Volume116
DOIs
StatePublished - Nov 5 2012

All Science Journal Classification (ASJC) codes

  • Radiation
  • Radiological and Ultrasound Technology
  • Biophysics
  • Radiology Nuclear Medicine and imaging

Keywords

  • Chelates
  • Labeling
  • Lanthanide
  • Luminescense
  • Multiple
  • Synthesis

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