Capillary monolithic bioreactor of immobilized snake venom phosphodiesterase for mass spectrometry based oligodeoxynucleotide sequencing.
A capillary monolithic bioreactor of snake venom phosphodiesterase (SVP) was constructed to generate different single-nucleotide mass ladders of oligodeoxynucleotides for mass spectrometry (MS)-based sequencing by immobilization. The immobilization of SVP in the porous silica monolith significantly enhances its stability for prolonged and repeated applications. The constructed capillary bioreactor has the advantages of handling (sub)microliter DNA samples and having good permeability. Benefiting from its good permeability, DNA solutions can be directly injected into the sequential digestion bioreactor simply by hand pushing or a low-pressure microinjection pump. Moreover, the immobilization of SVP facilitates the elimination or repression of the metal adducts of oligodeoxynucleotides, improving the analytical performance of MS sequencing. By the application of capillary bioreactor of immobilized SVP, the sequence-specific modification of single-stranded oligodeoxynucleotide induced by a ubiquitous pollutant acrolein (Acr) was identified, demonstrating its promising applications in identification of sequence-specific damage, which may further our understanding of DNA damage caused mutagenesis.
State Key Laboratory of Environmental Chemistry and Ecotoxicology, Research Center for Eco-Environmental Sciences, Chinese Academy of Sciences, Beijing 100085, China.
Chao
C
Zhao
Ruichuan
R
Yin
Junfa
J
Yin
Dapeng
D
Zhang
Hailin
H
Wang
D019149:Bioreactors; D019075:Electrophoresis, Capillary; D004800:Enzymes, Immobilized / Q000737:chemistry*; D009841:Oligonucleotides / Q000737:chemistry*; D043264:Phosphodiesterase I / Q000737:chemistry*; D017422:Sequence Analysis, DNA / Q000379:methods*; D012910:Snake Venoms / Q000201:enzymology; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*
D004800:Enzymes, Immobilized; D009841:Oligonucleotides; D012910:Snake Venoms; D043264:Phosphodiesterase I; C470790:phosphodiesterase I, snake venom