Galactosylated multimodular lipoplexes for specific gene transfer into primary hepatocytes.

Background: Numerous synthetic cationic vectors have been synthesized and are successfully used for in vitro gene transfer but an excess of positive charges can lead to cytotoxicity and does not enable specific transfection.

Methods: We decided to develop alternative molecular systems consisting of neutral, colloidally stable bioassemblies equipped with ligands for specific cell targeting. Consequently, we directed our efforts toward the development of a multimodular non-viral gene delivery system consisting of a condensed core of DNA with cationic liposomes of bis(guanidinium)-tren-cholesterol and an external corona of poly(ethylene oxide) stretches harbored by the steric stabilizers used to stabilize lipoplexes colloidally.

New methods for chemo-enzymatic galactosidation of 2S rapeseed protein.

Two chemo-enzymatic methodologies to synthesize neoglycoproteins from rapeseed 2S protein (napin) were developed. In the first approach, glycosidases were used to catalyse 1-O-glycosylation of serine residues, whereas in the second one, 6-N-galactosylation was examined using an amino-reduction reaction between the epsilon-NH2 of lysine residues and 6-oxogalactosides (readily available by means of the oxidation reaction of the corresponding galactosides mediated by galactose oxidase). Our results indicated that glycosidases were unable to glycosylate native proteins.

Alpha-galactosyl fluoride in transfer reactions mediated by the green coffee beans alpha-galactosidase in ice.

We show that the yields in saccharide synthesis by tranglycosylation with alpha-galactosidase from green coffee beans can be greatly enhanced when working in ice. Thus, methyl alpha-D-galactopyranosyl-(1-->3)-alpha-D-galactopyranoside (3a) produced by reaction of alpha-D-galactopyranosyl fluoride 1 with methyl alpha-D-galactopyranoside (2) is obtained with 51% yield in ice while only 29% is synthesized at 37 degrees C. This result, already previously found by others with proteases and by us with a beta-galactosidase appears to be a general property of hydrolases.