[Development of a method for methylated DNA enrichment with functionalized mesocellular silica foams immobilized with methyl CpG binding domain].
OBJECTIVE
To develop a method for enriching methylated DNA in clinical samples using mesocellular silica foams (MCFs) immobilized with methyl-CpG binding domain (MBD).


METHODS
MCFs with ultra-large pore size were synthesized, functionalized and immobilized with GST-MBD.


RESULTS
The large cage-like pore structures of MCF materials was retained after functionalization and immobilization, with pore diameter of 55 nm, window size of 30 nm, and a high pore volume of 1.0 cm(3)/g. The loading amount of MBD was as high as 53 wt%. Immobilized MBD showed high binding activity and stability. In a binding buffer with salt concentrations ranging 500-550 mmol/L, the MCF-MBD can selectively enrich methylated DNA from the mixed DNA solution.


CONCLUSIONS
The MCF-MBD method may offer a better choice for high-throughout DNA methylation screening, and has laid a foundation for clinical application, prenatal diagnosis and research on DNA methylation-related genetic diseases.
Key Laboratory of Molecular Medicine, Ministry of Education, Shanghai Medical College, Institute of Biomedical Science, Fudan University, Shanghai 200032, P. R. China.
Ya-ting
YT
Chen
Lu
L
Han
Dong-yuan
DY
Zhao
Bo
B
Tu
Duan
D
Ma
D000818:Animals; D018899:CpG Islands; D004247:DNA / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D019175:DNA Methylation; D004268:DNA-Binding Proteins / Q000737:chemistry*; D055767:Immobilized Proteins / Q000737:chemistry*; D017434:Protein Structure, Tertiary; D051381:Rats; D012822:Silicon Dioxide / Q000737:chemistry*
D004268:DNA-Binding Proteins; D055767:Immobilized Proteins; D012822:Silicon Dioxide; D004247:DNA