Mineral oil-, glycerol-, and Vaseline-coated plates as matrix-assisted laser desorption/ionization sample supports for high-throughput peptide analysis.
A novel protocol for rapid and high-quality sample preparation prior to matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) has been developed by coating bare stainless steel plates with one of three adhesives: mineral oil, glycerol, or Vaseline. The advantages of these three adhesive coats are that they take little time to both prepare and wipe away, hold the matrices to prevent them from flying from the support, reduce the background matrix, and affect neither the resolution of the peptide peaks nor the accuracy of their determined molecular masses. Consequently, the signal intensity, detection limit, and tolerance of the analytes to contaminants on the three adhesive-coated plates are improved. In the two strategies of on-plate desalting and concentration of the peptide mixture, all three adhesives reduced the loss of peptides, especially in the case of larger molecular mass peptides. The microscope and stereomicroscope images of the deposited droplets showed that after dropping onto the adhesive coats, the droplets formed a reduced spot size, were more homogeneous, and showed sticky crystallization. Therefore, this is an easy-to-use, reproducible, highly sensitive, tolerant (to salts), and high-throughput method of peptide sample preparation for MALDI-TOF MS analysis.
Zhuo
Hui-Qin
HQ
Key Laboratory of MOE for Cell Biology and Tumor Cell Engineering, School of Life Sciences, Xiamen University, Xiamen 361005, China.
Huang
Lin
L
Feng
Li-Jian
LJ
Huang
He-Qing
HQ
D000268:Adhesiveness / Q000187:drug effects; D000269:Adhesives; D002853:Chromatography, Liquid; D003911:Dextrans; D005990:Glycerol / Q000737:chemistry*; D006801:Humans; D008853:Microscopy; D008899:Mineral Oil / Q000737:chemistry*; D008970:Molecular Weight; D010455:Peptides / Q000032:analysis* / Q000097:blood / Q000737:chemistry; D010577:Petrolatum / Q000737:chemistry*; D012492:Salts / Q000494:pharmacology; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*; D013193:Stainless Steel
D000269:Adhesives; D003911:Dextrans; D010455:Peptides; D012492:Salts; D013193:Stainless Steel; D010577:Petrolatum; D008899:Mineral Oil; C025614:sephadex; D005990:Glycerol