Selective separation and enrichment of peptides for MS analysis using the microspheres composed of Fe3O4@nSiO2 core and perpendicularly aligned mesoporous SiO2 shell.
In this work, we report the development of a novel enrichment protocol for peptides by using the microspheres composed of Fe(3)O(4)@nSiO(2) Core and perpendicularly aligned mesoporous SiO(2) shell (designated Fe(3)O(4)@nSiO(2)@mSiO(2)). The Fe(3)O(4)@nSiO(2)@mSiO(2) microspheres possess useful magnetic responsivity which makes the process of enrichment fast and convenient. The highly ordered nanoscale pores (2 nm) and high-surface areas of the microspheres were demonstrated to have good size-exclusion effect for the adsorption of peptides. An increase of S/N ratio over 100 times could be achieved by using the microspheres to enrich a standard peptide, and the application of the microspheres to enrich universal peptides was performed by using myoglobin tryptic digest solution. The enrichment efficiency of re-used Fe(3)O(4)@nSiO(2)@mSiO(2) microspheres was also studied. Large-scale enrichment of endogenous peptides in rat brain extract was achieved by the microspheres. Automated nano-LC-ESI-MS/MS was applied to analyze the sample after enrichment, and 60 unique peptides were identified in total. The facile and low-cost synthesis as well as the convenient and efficient enrichment process of the novel Fe(3)O(4)@nSiO(2)@mSiO(2) microspheres makes it a promising candidate for selectively isolation and enrichment of endogenous peptides from complex biological samples.
Chen
Hemei
H
Department of Chemistry and Institutes of Biomedical Sciences, Fudan University, Shanghai, P R China.
Liu
Shasha
S
Yang
Huili
H
Mao
Yu
Y
Deng
Chunhui
C
Zhang
Xiangmin
X
Yang
Pengyuan
P
D000595:Amino Acid Sequence; D000804:Angiotensin II / Q000737:chemistry; D000818:Animals; D002417:Cattle; D045304:Cytochromes c / Q000378:metabolism; D052203:Ferrosoferric Oxide / Q000138:chemical synthesis / Q000737:chemistry*; D008863:Microspheres; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010455:Peptides / Q000737:chemistry / Q000302:isolation & purification*; D016062:Porosity; D051381:Rats; D012710:Serum Albumin, Bovine / Q000737:chemistry / Q000378:metabolism; D012822:Silicon Dioxide / Q000737:chemistry*; D012996:Solutions; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*
D010455:Peptides; D012996:Solutions; D000804:Angiotensin II; D012710:Serum Albumin, Bovine; D012822:Silicon Dioxide; D045304:Cytochromes c; D052203:Ferrosoferric Oxide