RAPID ANALYSIS OF BIOPOLYMERS ON MODIFIED NONPOROUS POLYSTYRENE - DIVINYLBENZENE PARTICLES

The use of short columns packed with alkylated non-porous 2.1 mum polystyrene - divinylbenzene particles has enabled the efficient separation of proteins, oligonucleotides, and DNA fragments in less than 60 seconds. An increase in the flow-rate resulted in only a minor reduction in the resolution of the proteins. Careful temperature control and small gradient dead volume were essential if reproducible quantitative results were to be achieved. Analysis at elevated temperatures not only reduced the viscosity of the eluents and, hence, column back-pressure but also resulted in higher column efficiency. The best resolution of proteins was achieved by performing the separation at 80-degrees-C whereas the optimum temperatures for the separation of oligonucleotides and DNA fragments ranged from 40-50-degrees-C. High speed analysis was used both to control the purity of oligonucleotides following automated solid-phase synthesis and to evaluate the expression of multidrug resistance genes in patients suffering from chronic lymphatic leukemia.