Highly bright avidin-based affinity probes carrying multiple lanthanide chelates

Document Type

Article

Publication Date

11-5-2012

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. © 2012 Elsevier B.V. All rights reserved.

Identifier

84865478975 (Scopus)

Publication Title

Journal of Photochemistry and Photobiology B Biology

External Full Text Location

https://doi.org/10.1016/j.jphotobiol.2012.07.001

e-ISSN

18732682

ISSN

10111344

PubMed ID

23018156

First Page

22

Last Page

29

Volume

116

Grant

RO1 GM-307-17-21

Fund Ref

National Institutes of Health

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