Merck Sharp + Dohme Corp.

Schering Corporation

Schering-Plough Corporation

Susan V Cannon-Carlson

Seoju Lee

David C Wylie

Gary J Vellekamp

Collette Marie Cutler

John C Tang

Marcio Voloch

Andres Frei

Brittany Charlotte Larkin

Roland Mengisen

Kimberly Margaret Louise May

Andres c/o Schering-Plough Corporation Frei

Interleukin-10 (IL-10) conjugated via a linker to one or more polyethylene glycol (PEG) molecules at a single amino acid residue of the IL-10, and a method for preparing the same, are provided. The method produces a stable mono-pegylated IL-10, which retains IL-10 activity, where pegylation is selective for the N-terminus on one subunit of IL-10 with little or no formation of monomeric IL-10. The method also provides a substantially homogenous population of mono-PEG-IL-10.

Pegylated interleukin-10

The present invention includes a method for reducing the appearance of false positive bands in SDS-PAGE analysis of proteolytic digestion, of a sample that comprises an immunoglobulin polypeptide, at pH 8.8.

Method for reducing the appearance of false positive bands in SDS-page analysis of proteolytic digestion of a sample

Methods for purifying adenoviruses from a contaminated sample using a hydroxyapatite medium are provided. Sodium chloride concentrations present in buffers used throughout the method are at least 150 mM to prevent the adenovirus from irreversibly binding to the hydroxyapatite. Levels of contaminants and empty adenovirus capsids from samples purified by conventional purification techniques are further reduced to provide a highly pure adenovirus preparation.

Method for purifying adenoviruses

The present invention provides methods for isolating adjunct isoforms of proteins and converting them to the desired protein. In preferred embodiments, the present invention contemplates the use of acid solutions or zinc solutions to cleave a chemical group from a desired protein. In further preferred embodiments, the present invention contemplates the oxidation of reduced sulfhydryl groups with cleavage of chemical groups to form a functional desired protein.

Methods for conversion of protein isoforms

Purification of bacterially expressed human interleukin-10

A method for purifying Interleukin-10 (Il-10) from a eukaryotic cell culture medium. The method is comprised of subjecting IL-10 containing cell culture medium to cation exchange chromatography, anion exchange chromatography, hydroxyapatite chromatography, and gel filtration chromatography. The present invention is also comprised of a process for separating different IL-10 dimers present in a protein fraction from each other by subjecting the protein fraction to hydroxyapatite chromatography. The present invention is also comprised of a process for separating variants of a protein differing in an N-terminal amino acid sequence present in a protein fraction from each other by subjecting the protein fraction to hydroxyapatite chromatography.

Purification of human interleukin-10 from a cell culture medium

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Wayne, NJ, United States of America

Susan V Cannon-Carlson