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
Recommended Citation
Wirpsza, Laura; Pillai, Shyamala; Batish, Mona; Marras, Salvatore; Krasnoperov, Lev; and Mustaev, Arkady, "Highly bright avidin-based affinity probes carrying multiple lanthanide chelates" (2012). Faculty Publications. 18035.
https://digitalcommons.njit.edu/fac_pubs/18035
