Construction of a high-performance magnetic enzyme nanosystem for rapid tryptic digestion.
A magnetic enzyme nanosystem have been designed and constructed by a polydopamine (PDA)-modification strategy. The magnetic enzyme nanosystem has well defined core-shell structure and a relatively high saturation magnetization (Ms) value of 48.3 emu g(-1). The magnetic enzyme system can realize rapid, efficient and reusable tryptic digestion of proteins by taking advantage of its magnetic core and biofunctional shell. Various standard proteins (e.g. cytochrome C (Cyt-C), myoglobin (MYO) and bovine serum albumin (BSA)) have been used to evaluate the effectiveness of the magnetic enzyme nanosystem. The results show that the magnetic enzyme nanosystem can digest the proteins in 30 minutes, and the results are comparable to conventional 12 hours in-solution digestion. Furthermore, the magnetic enzyme nanosystem is also effective in the digestion of low-concentration proteins, even at as low as 5 ng μL(-1) substrate concentration. Importantly, the system can be reused several times, and has excellent stability for storage. Therefore, this work will be highly beneficial for the rapid digestion and identification of proteins in future proteomics.
Department of Biomedical engineering, The Pennsylvania State University, University Park, PA 16802, (USA).
Gong
G
Cheng
Department of Biomedical engineering, The Pennsylvania State University, University Park, PA 16802, (USA).
Si-Yang
SY
Zheng
D000818:Animals; D002417:Cattle; D045304:Cytochromes c / Q000737:chemistry; D004795:Enzyme Stability; D017743:Equipment Reuse; D052203:Ferrosoferric Oxide / Q000737:chemistry*; D007211:Indoles / Q000737:chemistry*; D059346:Magnets; D009211:Myoglobin / Q000737:chemistry; D049329:Nanostructures / Q000737:chemistry* / Q000648:ultrastructure; D011108:Polymers / Q000737:chemistry*; D059748:Proteolysis; D040901:Proteomics / Q000295:instrumentation* / Q000379:methods; D012710:Serum Albumin, Bovine / Q000737:chemistry; D014357:Trypsin / Q000737:chemistry*
D007211:Indoles; D009211:Myoglobin; D011108:Polymers; C568283:polydopamine; D012710:Serum Albumin, Bovine; D045304:Cytochromes c; D014357:Trypsin; D052203:Ferrosoferric Oxide