Trypsin-immobilized fiber core in syringe needle for highly efficient proteolysis.
In this report, a core-changeable needle enzymatic reactor was developed for highly efficient proteolysis. A piece of enzyme-immobilized fiber core was inserted into the needle of a syringe pump to form a flow-through bioreactor. The novel in-needle bioreactor could be regenerated by changing its fiber core. The feasibility and performance of the unique bioreactor were demonstrated by the tryptic digestion of BSA and lysozyme and the digestion time was significantly reduced to less than 5 s. The digests were identified by MALDI-TOF MS with sequence coverages comparable to those obtained by the conventional in-solution tryptic digestion. The present in-needle bioreactor provides a promising platform for the high-throughput protein identification.
Wang
Sheng
S
School of Pharmacy, Department of Chemistry, Fudan University, Shanghai, China.
Chen
Zhi
Z
Yang
Pengyuan
P
Chen
Gang
G
D000818:Animals; D019149:Bioreactors; D002241:Carbohydrates / Q000737:chemistry; D002417:Cattle; D046208:Microchip Analytical Procedures; D009113:Muramidase / Q000737:chemistry*; D009339:Needles; D010449:Peptide Mapping; D040901:Proteomics / Q000379:methods*; D012710:Serum Albumin, Bovine / Q000737:chemistry*; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*; D013594:Syringes; D013997:Time Factors; D014357:Trypsin / Q000737:chemistry*
D002241:Carbohydrates; D012710:Serum Albumin, Bovine; D009113:Muramidase; D014357:Trypsin