Optimized conditions to couple two water-soluble biomolecules through alkylamine thiolation and thioetherification

Citation
L. Meunier et al., Optimized conditions to couple two water-soluble biomolecules through alkylamine thiolation and thioetherification, BIOCONJ CHE, 10(2), 1999, pp. 206-212
Citations number
22
Categorie Soggetti
Chemistry & Analysis
Journal title
BIOCONJUGATE CHEMISTRY
ISSN journal
10431802 → ACNP
Volume
10
Issue
2
Year of publication
1999
Pages
206 - 212
Database
ISI
SICI code
1043-1802(199903/04)10:2<206:OCTCTW>2.0.ZU;2-Q
Abstract
A simple method far introducing, in buffered saline, a reactive sulfhydryl group on water-soluble molecules bearing an alkyl-amino group is described. This method is based on the use of two water-soluble reagents: 2-iminothio lane and 6,6'-dithiodinicotinic acid. The first one is open upon reaction w ith an amino group, and the generated thiol group is immediately protected by action of the second reagent. The optimal conditions were determined by taking into account the stability and the reactivity of both reagents with regards to pH and temperature. This method was validated through two applic ations, the substitution of bovine serum albumin with a bromoacetyl peptide and the substitution of an amino link at the 5' end of an oligonucleotide by reaction with either a fluorescent tag, iodoacetamidofluorescein, or a b romoacetyl peptide, upon reduction of the protected disulfide bridge with a third water-soluble reagent, namely tris(2-carboxyethyl)phosphine.