Virginia Tech
Delipidation Treatments for Large-Scale Protein Purification Processing
Abstract
dc:description.abstractTriglycerides are the majority lipid component of most biochemical mixtures and are virtually water insoluble. Lipid removal is desired prior to protein purification processing to decrease nonspecific fouling of downstream chromatographic matrices. Transgenic pig milk was used as a model system to study delipidation from therapeutic protein sources. The majority of triglycerides was extracted from stable lipid micelles and removed with a method that can be incorporated in downstream protein purification processing without denaturing the target protein. An efficient delipidation treatment used TNBP, a non-polar solvent, to extract lipid micelles and then phase transfer milk lipids into a TNBP-swelled dextran particulate. A batch incubation of a whey/TNBP mixture with pre-swollen Sephadex LH-20 or hydroxyalkoxypropyl dextran (HAPD) beads at 4 C for 24 hours removed 67 + 2 % (0.645 mg triglycerides/ml Sephadex LH-20) and 71 o + 1 % (0.628 mg triglycerides/ml HAPD) of the triglycerides present in the skimmed transgenic whey, respectively. Fully swollen beads removed 20% more triglycerides than beads which were wetted but not swollen in TNBP, indicating that a larger phase volume and internal adsorption of the lipids onto the Sephadex matrix dominates over surface adsorption. Polyclonal ELISAs indicated that 89 + 6% of the recombinant human Protein C was still present in the transgenic whey after this delipidation treatment, indicating this treatment did not denature or harm the target protein.
Degree
thesis:*- Name thesis:degree_name
- Master of Science
- Level thesis:degree_level
- masters
- Discipline thesis:degree_discipline
- Chemical Engineering
- Department dc:contributor.department
- Chemical Engineering
- Grantor dc:publisher
- Virginia Tech
- Year dc:date.issued
- 1998
Author and committee
dc:creator, dc:contributor.*- Author dc:creator
-
- Gardner, Tara Conti
- Chair dc:contributor.committeechair
-
- Velander, William H.
- Committee members dc:contributor.committeemember
-
- Davis, Richey M.
- Conger, William L.
Subjects
dc:subject × 6Rights
dc:rights- Statement dc:rights
-
- In Copyright
- Licence dc:rights.uri
Identifiers
dc:identifier.*- Dc Identifier Other
- etd-1454132679612381
- OAI identifier oai:identifier
- oai:vtechworks.lib.vt.edu:10919/36512