| Type of Document |
Dissertation |
| Author |
Degener, Jr., Arthur W.
|
| Author's Email Address |
adegener@vt.edu |
| URN |
etd-042399-164339 |
| Title |
Downstream processing of recombinant and endogenous proteins from livestock milk
|
| Degree |
PhD |
| Department |
Chemical Engineering |
| Advisory Committee |
| Advisor Name |
Title |
| Dr. William Velander |
Committee Chair |
| Dr. Kimberly Foresten |
Committee Member |
| Dr. Richey Davis |
Committee Member |
| Dr. William Conger |
Committee Member |
| Dr. William Drohan |
Committee Member |
|
| Keywords |
- endogenous milk proteins
- recombinant proteins
- free flow iso-electric focuing
- Free flow electrophoresis
- Expanded bed adsorption chromatography
|
| Date of Defense |
1999-04-08 |
| Availability |
unrestricted |
Abstract
With the increased demands of therapeutic proteins, there is going to be a need for new purification technologies which have high throughput, high yield and high resolution. Three purification technologies were explored as potential new technology to isolate recombinant and endogenous milk proteins: Expanded bed adsorption chromatography(EBAC) combined with hydrophobic interaction chromatography(HIC), Recycle continuous flow electrophoresis(RCFE) and Free flow isoelectric focusing(FFIEF). 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. The second process(RCFE) was able to isolate the active sub-populations of rhPC from major milk contaminants( - and -pig casein) as wells as from the inactive sub-populations of rhPC. RCFE was able to process 1.5g total protein per hour on a small scale and is currently being researched to process 1kg total protein per hour. The third and final purification process(FFIEF) sub-fractionated 100mg of immuno-purified rhPC into 50 fractions. The FFIEF was able to produce a linear pH gradient over the range of 3-10 using 2% ampholytes. The fractionated rhPC showed differing degrees of activity that resulted from the -carboxylated glutamic acids and the sialic acids.
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| Files |
| Filename |
Size |
Approximate Download Time
(Hours:Minutes:Seconds)
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| 28.8 Modem |
56K Modem |
ISDN (64 Kb) |
ISDN (128 Kb) |
Higher-speed Access |
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thesis.PDF |
2.07 Mb |
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