pr070132s_si_001.pdf (62.78 kB)
Immobilization of Trypsin on Superparamagnetic Nanoparticles for Rapid and Effective Proteolysis
journal contribution
posted on 2020-04-02, 15:50 authored by Yan Li, Xiuqing Xu, Chunhui Deng, Pengyuan Yang, Xiangmin ZhangIn this work, a novel and facile route was developed for the immobilization of enzyme on nanosized
magnetic particles, and its application to fast protein
digestion via a direct MALDI-TOF mass spectrometry
analysis was demonstrated. At first, amine-functionalized
magnetic particles with high magnetic responsivity and
excellent dispersibility were prepared through a facile
one-pot strategy. Then, magnetic nanoparticles were
functionalized with numerous aldehyde(−CHO) groups by
treating the as-synthesized, amine-functionalized magnetic nanoparticles with glutaraldehyde. Finally, immobilization of trypsin onto the aldehyde-functionalized
magnetic nanoparticles was achieved through reaction of
the aldehyde groups with amine groups of trypsin. The
obtained trypsin-immobilized magnetic nanoparticles were
conveniently applied for protein digestion. The digestion
efficiency was demonstrated with peptide mapping analysis of three model proteins. The process of digestion is
very facile due to the easy manipulation of magnetic
nanoparticles. Complete protein digestion was achieved
in a short time (5 min), without any complicated reduction
and alkylation procedures. These results are expected to
open up a new possibility for the proteolysis analysis as
well as a new application of magnetic nanoparticles.
Additionally, it is worth noting that, since the preparation
and surface functionality of magnetic nanoparticles is low-cost and reproducible, the preparation method and application approach of the magnetic nanoparticles may find
much potential in proteome research.
Keywords: amine-functionalized magnetic particles • immobilized trypsin • digestion on magnetic nanoparticles