Abstract

Luminescent lanthanide chelates have been used to label antibodies in time-gated luminescence (TGL) bioimaging. However, it is a challenging task to label directly an antibody with lanthanide-binding ligands and achieve control of the target ligand/protein ratios whilst ensuring that affinity and avidity of the antibody remain uncompromised. We report the development of a new indirect detection reagent to label antibodies with detectable luminescence that circumvents this problem by labelling available lysine residues in the linker portion of the recombinant fusion protein Linker-Protein G (LPG). Succinimide-activated lanthanide chelating ligands were attached to lysine residues in LPG and Protein G (without Linker) and the resulting Luminescence-Activating (LA-) conjugates were compared for total incorporation and conjugation efficiency. A higher and more efficient incorporation of ligands at three different molar ratios was observed for LPG and this effect was attributed to the presence of eight readily available lysine residues in the linker region of LPG. These Luminescence-Activating (LA-) complexes were subsequently shown to impart luminescence (upon formation of europium(III) complexes) to cell-specific antibodies within seconds and without the need for any complicated bioconjugation procedures. The potential of this technology was demonstrated by direct labelling of Giardia cysts and Cryptosporidium oocysts in TGL bioimaging.

LanguageEnglish
Article number27564
Pages1-9
Number of pages9
JournalScientific Reports
Volume6
DOIs
Publication statusPublished - 10 Jun 2016

Fingerprint

Luminescence
Lanthanoid Series Elements
Lysine
Ligands
Antibody Affinity
Antibodies
Proteins
Europium
Recombinant Fusion Proteins
Giardia
Cryptosporidium
Oocysts
GTP-Binding Proteins
Cysts
Technology

Bibliographical note

Version archived for private and non-commercial use with the permission of the author/s and according to publisher conditions. For further rights please contact the publisher.

Cite this

Sayyadi, Nima ; Care, Andrew ; Connally, Russell E. ; Try, Andrew C. ; Bergquist, Peter L. ; Sunna, Anwar. / A novel universal detection agent for time-gated luminescence bioimaging. In: Scientific Reports. 2016 ; Vol. 6. pp. 1-9.
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A novel universal detection agent for time-gated luminescence bioimaging. / Sayyadi, Nima; Care, Andrew; Connally, Russell E.; Try, Andrew C.; Bergquist, Peter L.; Sunna, Anwar.

In: Scientific Reports, Vol. 6, 27564, 10.06.2016, p. 1-9.

Research output: Contribution to journalArticleResearchpeer-review

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AU - Bergquist,Peter L.

AU - Sunna,Anwar

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