Novel method for coupling of poly(ethyleneglycol) to carboxylic acid moieties of proteins.
Lack of toxicity, excellent solubility and superb biocompatibility make polyethylene glycol (PEG) one of the most popular modifiers of biologicals. The most common method for attachment of PEG is based on modification of amino groups of proteins with methoxy- or succinimide-derivatives of PEG. In the case of proteins with amino groups important for biological activity, this modification can lead to inactivation of proteins. A new strategy for covalent attachment of PEG to carboxylic groups of proteins using O,O-bis-(2-aminopropyl)polyethylene glycol and carbodiimide/N-hydroxysuccinimide-mediated reaction was developed. The reaction is carried out under mild aqueous conditions. The attached PEG serves as a hydrophilic spacer for further bioconjungation with biomolecules and haptens. Lipase from Candida rugosa was used as a model protein. Characteristic data of the modified protein such as activity, isoelectric points and stability were compared with that of the unmodified protein.
Kynclova
E
E
Institut für Biochemie und Molekulare Zellbiologie, University of Vienna, Wien, Austria.
Elsner
E
E
Köpf
A
A
Hawa
G
G
Schalkhammer
T
T
Pittner
F
F
D001672:Biocompatible Materials / Q000737:chemistry / Q000378:metabolism*; D002175:Candida / Q000201:enzymology; D002264:Carboxylic Acids / Q000378:metabolism*; D005656:Fungal Proteins / Q000737:chemistry / Q000378:metabolism; D008049:Lipase / Q000737:chemistry / Q000378:metabolism; D011092:Polyethylene Glycols / Q000737:chemistry / Q000378:metabolism*; D011487:Protein Conformation; D011506:Proteins / Q000378:metabolism*
D001672:Biocompatible Materials; D002264:Carboxylic Acids; D005656:Fungal Proteins; D011506:Proteins; D011092:Polyethylene Glycols; D008049:Lipase