Site-specific conjugation of the quencher on peptide's N-terminal for the synthesis of a targeted non-spreading activatable optical probe

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DOIResolve DOI: http://doi.org/10.1002/psc.2884
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TypeArticle
Journal titleJournal of Peptide Science
ISSN1075-2617
Volume22
Issue6
Pages415420
SubjectN-terminal; Site-specific; Conjugation; Quencher; Fluorescence; Targeted; Smart probe; Activatable
AbstractOptical imaging offers high sensitivity and portability at low cost. The design of ‘smart’ or ‘activatable’ probes can decrease the background noise and increase the specificity of the signal. By conjugating a fluorescent dye and a compatible quencher on each side of an enzyme's substrate, the signal remains in its ‘off ’ state until it reaches the area where a specific enzyme is expressed. However, the signal can leak from that area unless the dye is attached to a molecule able to bind to a specific target also presented in that area. The aim of this study was to (i) specifically conjugate the quencher on the α-amino group of the peptide's N-terminus, (ii) conjugate the dye on the ε-amino group of a lysine in C-terminus, and (iii) conjugate the carboxyl group of the peptide's C-terminus to an amino group present on an antibody, using carbodiimide chemistry. The use of protecting groups, such as Boc or Fmoc, to allow site-specific conjugation, presents several drawbacks including ‘on beads labeling’, additional steps required for deprotection and removal from the resin, decreased yield, and dye degradation. A method of preferential labeling of α-amino N-terminal group in slightly acidic solution, proposed by Selo et al. (1996) has partially solved the problem. The present study reports improvements of the method allowing to (i) avoid the homo-bilabeling, (ii) increase the yield of the N-terminal labeling by two folds, and (iii) decrease the cost by 44-fold.
Publication date
PublisherWiley
LanguageEnglish
AffiliationHuman Health Therapeutics; National Research Council Canada
Peer reviewedYes
NPARC number23000285
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Record identifierf14b18a6-e1df-4d95-b2a3-89d99dcc8560
Record created2016-07-04
Record modified2016-07-04
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