Downstream Processing of Recombinant and EndogenousProteins from Livestock Milk

byArthur W. Degener, Jr.Faculty of the Virginia Polytechnic Institute and State University

Background

Introduction

Introduction

With the Increased demands of therapeutic proteins, there is going to be a need for new purification technologies which have

high throughputhigh yield

high resolution

Introduction

Three purification technologies were explored as potential new technology to isolate recombinant and endogenous milk proteins

1. Expanded bed adsorption chromatography(EBAC) combined with hydrophobic interaction chromatography(HIC)

2. Recycle continuous flow electrophoresis(RCFE) 3. Free flow isoelectric focusing(FFIEF).

Introduction

The first process(EBAC/HIC) used with Zn2+ as a selective precipitating agent, purified recombinant human protein C (rhPC) and IgG (contaminated with less than 1% IgA) from swine milk with high resolution and high yield while processing about 10-20 grams in a single operation.

1. Expanded bed adsorption chromatography(EBAC)

Dynamic light scattering (DLS) measurements

Dynamic light scattering (DLS) measurements

2. Hydrophobic interaction chromatography(HIC)

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