An investigation of the preparation of insoluble cellulose sulphates and their use for the isolation of lipoproteins from serum : a thesis presented in partial fulfilment of the requirements for the degree of Master of Science in Chemistry at Massey University, New Zealand
The aim of this thesis was to prepare an insoluble sulphated polysaccharide and explore its potential as a rapid column chromatographic method for the isolation and separation of lipoproteins directly from serum. Regenerated cellulose crosslinked with formaldehyde was found to be relatively inert towards esterification. A degree of sulphation of 0.5 milliequivalents per gram could be obtained but the product had poor hydraulic properties. Epichlorohydrin crosslinked regenerated cellulose was found to be readily sulphated so long as the matrix was highly crosslinked (50-100% v/w). The reactivity of the matrix was retained at low levels of crosslinking if hydroxypropyl groups were introduced with propylene oxide. The hydroxypropylated regenerated celluloses were readily sulphated up to four milliequivalents per gram with minimal degradation of the cellulose chain. HP-cellulose 8-50 sulphates proved to be very successful in the selective isolation of the beta lipoproteins (VLDL plus LDL) directly from serum over a wide range of conditions. The conditions necessary for the preparation of pure alpha lipoprotein (HDL) were found to be more critical. Employing conditions so that all of the alpha lipoprotein (HDL) was sequested completely from serum resulted in it being contaminated by other serum proteins. Under other conditions where the alpha lipoprotein (HDL) was obtained without contamination by other serum proteins, it appeared that a small sub-fraction of the alpha lipoprotein remained in the serum.