title: Enzyme-linked bridging assay method for the quantification of oligonucleotide-based drugs in biological matrices.
abstract: With ongoing efforts to develop oligonucleotide-based (ODN-based) therapeutics, there is a need for a sensitive, high-throughput method of quantification of ODN-based drugs in biological matrices. To overcome the insufficient sensitivity and time-consuming sample extraction procedures involved in conventional capillary gel electrophoresis (CGE) and high-performance liquid chromatography (HPLC), we developed a nucleic acid hybridization-based enzyme-linked bridging assay (ELBA), which shows significant advantages over CGE methods in evaluating ODN-based drugs in plasma and tissue: (1) It has higher sensitivity; (2) it involves easier sample extraction procedures; (3) it is suitable for many ODN-based drugs, even those with different secondary structures and modifications, including phosphorothioate oligonucleotide (PSODN), mixed backbones with 2'-O-Me (MBO), locked nucleic acid (LNA) modifications, and B- and C-type CpG sequences; and (4) it is highly selective, even during simultaneous quantification, with regard to intact ODNs and their 3'-metabolites. This universal design produces a rapid, sensitive, specific assay with minimal method development time. It is well suited to high-throughput analysis of various ODN-based drugs.
authors: Department of Pharmacology and Toxicology, Beijing Institute of Radiation Medicine, Beijing, People's Republic of China.
authors: Jie
authors: J
authors: Fu
authors: Yunjuan
authors: Y
authors: Sun
authors: Shaoyou
authors: S
authors: Xia
authors: Lihou
authors: L
authors: Dong
authors: Qingqing
authors: Q
authors: Wang
authors: Lun
authors: L
authors: Ou
authors: Xianfei
authors: X
authors: Shen
authors: Zhiguo
authors: Z
authors: Lv
authors: Haifeng
authors: H
authors: Song
mesh_terms: D000818:Animals; D057075:Enzyme Assays / Q000295:instrumentation* / Q000379:methods; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D005260:Female; D000882:Haplorhini; D057166:High-Throughput Screening Assays / Q000379:methods; D008297:Male; D009690:Nucleic Acid Conformation; D009693:Nucleic Acid Hybridization / Q000379:methods*; D009841:Oligonucleotides / Q000032:analysis* / Q000097:blood / Q000737:chemistry; D004364:Pharmaceutical Preparations / Q000032:analysis*; D051381:Rats; D017208:Rats, Wistar; D012680:Sensitivity and Specificity
chemical_list: D009841:Oligonucleotides; D004364:Pharmaceutical Preparations