Optimization of large beaded cellulose as a chromatographic support
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The amount of immobilized antibody was varied and immunosorptive capacity of 1 mm cellulose beads was found to be equivalent to that of 0.1 mm cross~linked agarose beads. The immobilization of antibodies to these supports was studied by photomicroscopy of cross-sectioned beads containing immobilized fluorescent labeled antibodies. While 75% of the antibody was immobilized within 0.07 mm of the cellulose bead surface at an antibody density of 1 mg antibody per ml of beads, an appreciable amounts of antibody immobilized deeper into the bead may have been utilized in order to yield capacities equivalent to the smaller agarose beads. The beaded cellulose supports derivatized to form either immunoaffinity or anion exchange matrices exhibited very low non-specific binding. Thus, the particle size, solids content, and extent of derivatization of cellulose matrices can be engineered so as to create matrices that provide high flow rates with low pressure drops while also having desirable adsorptive capacity for proteins.
- Doctoral Dissertations