Postsynthetic conjugation of biopolymers with high molecular mass poly(ethylene glycol): Optimization of a solution process tested on synthetic oligonucleotides

The reaction of oligonucleotides with high molecular weight monomethoxy poly(ethylene glycol)s (MPEGs) has been tested to set up a convenient procedure for the postsynthetic conjugation in solution of biopolymers. A first oligonucleotide was previously modified in 5', using a liquid-phase procedure, with a linker carrying a terminal primary amino group to enhance its nucleophilic reactivity. Two procedures commonly utilized for the activation of the terminal OH groups of the MPEG were evaluated, that is, the reaction with pNO(2)-phenyl chloroformate and with N,N-disuccinimidyl carbonate. Both water as well as organic solution conditions were employed and compared. In a second test, a X-amino modified, commercial 20-mer was also conjugated in a microscale condition to verify the effect of size and concentration of MPEG on the postsynthetic conjugation of these biopolymers under troublesome synthetic conditions.