APPLICATION OF SAMPLE DISPLACEMENT TECHNIQUES TO THE PURIFICATION OF SYNTHETIC OLIGONUCLEOTIDES AND NUCLEIC-ACIDS - A MINIREVIEW WITH EXPERIMENTAL RESULTS

Clinical development of antisense drugs is rapidly increasing requirem ents for large quantities of oligonucleotides. As part of the manufact uring process appropriately large scale separation techniques must be developed. This article reviews applications of sample self displaceme nt techniques to the purification of oligonucleotides and nucleic acid s, wherein sample components act as displacers and enhance purities. A pplications for other solutes are also reviewed where similar methods may be of use in oligonucleotide purification. Experimental data is pr esented for sample self displacement anion-exchange chromatography of the clinically important phosphorothioate oligonucleotides. The techni que can enhance full-length purity of the product and offers maximal u tilization of the chromatographic media. Full-length purities of 95% a nd higher are achieved by this approach and recovery yields are up to 70% depending upon starting feed purity. Recycling and real time proce ss monitoring can further improve recovery. Techniques for column satu ration with multiple linked columns are presented and their applicatio ns to oligonucleotide purification are discussed. (C) 1998 Elsevier Sc ience B.V.