Protein conjugates for trypsin mediated pegylation by transamidation and methods

Herein is reported a method for producing of a polypeptide conjugated to one poly (ethylene glycol) comprising a) providing a nucleic acid encoding an expression construct comprising in 5′ to 3′ direction a nucleic acid encoding a polypeptide, and a nucleic acid encoding a trypsin site of SEQ ID NO:...

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Hauptverfasser: HESSE, FRIEDERIKE, MUELLER, STEPHANIE, TROST-GROSS, EVA MARIA, HOESS, EVA
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MUELLER, STEPHANIE
TROST-GROSS, EVA MARIA
HOESS, EVA
description Herein is reported a method for producing of a polypeptide conjugated to one poly (ethylene glycol) comprising a) providing a nucleic acid encoding an expression construct comprising in 5′ to 3′ direction a nucleic acid encoding a polypeptide, and a nucleic acid encoding a trypsin site of SEQ ID NO: 01, b) expressing the nucleic acid of a) in a cell and recovering the expression construct from the cell and/or the cultivation medium, c) providing a target peptide with an amino acid sequence of SEQ ID NO: 02 covalently conjugated to a poly (ethylene glycol) at the C-terminal lysine residue, d) incubating the expression construct and the target peptide with the trypsin mutant D189K, K60E, N143H, E151H, and e) recovering and thereby producing the polypeptide conjugated to one poly (ethylene glycol) from the incubation mixture.
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language eng ; fre ; ger
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subjects BEER
BIOCHEMISTRY
CHEMISTRY
ENZYMOLOGY
FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIREDCHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERSFROM A RACEMIC MIXTURE
HUMAN NECESSITIES
HYGIENE
MEDICAL OR VETERINARY SCIENCE
METALLURGY
MICROBIOLOGY
MUTATION OR GENETIC ENGINEERING
ORGANIC CHEMISTRY
PEPTIDES
PREPARATIONS FOR MEDICAL, DENTAL, OR TOILET PURPOSES
SPIRITS
VINEGAR
WINE
title Protein conjugates for trypsin mediated pegylation by transamidation and methods
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