Development and validation of a capillary electrophoresis method for the characterization of herpes simplex virus type 1 (HSV-1) thymidine kinase substrates and inhibitors

A highly sensitive capillary electrophoresis method using p-nitrophenyl 5′-thymidine monophosphate as a substrate for the monitoring of nucleotide pyrophosphatase/phosphodiesterase activities

“…Thus, the buffer combination phosphate-HEPES was chosen for the subsequent assay development. The ionic strength of the running buffer is also an important factor to control the separation efficiency and migration times [27,43]. Initially, a 50 mM concentration of running phosphate buffer was selected; however the internal standard did not separate from pnitrophenol under these conditions.…”

Identification and characterization of acyclovir-resistant clinical HSV-1 isolates from children

“…Genotypic tests were mainly performed through direct sequencing of tk and DNA pol to determine viral susceptibility based on databases of mutations related to ACV resistance. 5,7,8,[12][13][14][15][16] On the other hand, phenotypic evaluations, including plaque reduction assay (PRA), 17 dye uptake assay, and virus yield reduction assay, [18][19][20][21][22] were largely conducted on HSV infections that induced the cytopathic effect. These assays are time consuming, laborious, and hence inconvenient for routine clinical screening.…”

A capillary electrophoresis method with dynamic pH junction stacking for the monitoring of cerebroside sulfotransferase

“…Our group has long-standing experience in the quantification of nucleotides by CE, and especially in enzyme assays involving the formation or transformation of nucleotides [15][16][17][18][19][20][21][22]. Thus we decided to quantify the nucleotide PAP which is formed from the co-substrate PAPS during the CST reaction.…”