The need for high-purity oligodeoxyribonucleotides for various applications has resulted in the development of novel synthesis, purification, and analytical techniques. A diversity of methods, including polyacrylamide slab gel electrophoresis, capillary gel electrophoresis, as well as high-performance liquid chromatography (HPLC), have been successfully used to aid in the characterization and isolation of these synthetic compounds. The information contained in this review article primarily details both the theoretical and practical aspects related to the use of HPLC for the analysis and purification of synthetic DNA. In addition, a variety of postsynthesis sample preparation protocols, commonly employed prior to and after HPLC, are described.

The use of a rigid silica-based packing material with large particle and pore size, 37–55 μm and 500 Å pore, for affinity chromatography makes it possible to combine high selectivity with short analysis times. Both large and small molecules have been covalently bonded to the Protein-PakTM Affinity Epoxy-Activated bulk packing for purification of glycoproteins, immunoglobulins, enzymes, lectins and other proteins. Recombinant protein A, GammaBindTM G, heparin, Cibacron Blue F3G-A, sulfanilamide, N-acetyl-D-glucosamine, concanavalin A and aminophenylboronic acid were covalently attached to the affinity packing for selective purification of proteins.

ABSTRACT: We compared oxygen-related prostaglandin synthesis in fetal lamb ductus arteriosus (DA) pulmonary artery (PA) and aorta endothelial and smooth muscle cells. We measured basal synthesis of 6-keto-PGF1α and PGE2, the response to calcium ionophore (A23187), a nonspecific stimulus of prostaglandin production, as well as the response to oxygen, a perinatal stimulus, monitoring both the effects of hyperoxia (95% O2) and hypoxia (2% O2). In addition, we established whether differences observed in fetal lamb PA cells related to oxygen tension were also observed in newborn central and microvessel PA cells. Our results indicate that DA endothelial cells increase 6-keto- PGF1α in response to ionophore (p<0.05). With hyperoxia, DA endothelial cells increase PGE2 synthesis and DA smooth muscle cells increase 6-keto-PGFla (p<0.05 and 0.02, respectively). Aorta smooth muscle cells increase 6-keto-PGFia in response to ionophore and hyperoxia (p < 0.003 and 0.05, respectively). PA endothelial and smooth muscle cells have higher levels of basal prostaglandin synthesis when compared with DA and aorta. In response to ionophore, increased 6-keto-PGF]a is observed in both PA endothelial and smooth muscle cells (p<0.02 and 0.0004, respectively), and PGE2 is increased in PA smooth muscle cells (p<0.003). Hypoxia, however, decreases PA smooth muscle production of both 6-keto-PGFia and PGE2 (p<0.02 and 0.01, respectively). Similar observations were made in newborn lamb central and microvessel PA cells. Thus, the hyperoxia-induced increase in prostaglandin synthesis in DA (endothelium and smooth muscle) and aorta (smooth muscle) and the hypoxia-induced decrease in prostaglandin synthesis in PA (smooth muscle) seem to be phenotypic properties of the cells in culture.