Preparation method of universal chip for detection of protein and nucleic acid by time-of-flight mass spectrometry

Abstract

The invention provides a preparation method of a general chip for flight time mass spectrometry detection of protein and nucleic acid. The method comprises the steps as follows: (1) a chip with tough texture and good flatness is selected as a substrate or chip body; (2) the chip surface is subjected to hydrophilic treatment; (3) hydrophilic sample application holes are closed with a photoresist; (4) the chip is treated with a silane coupling agent selected from vinylsilane, aminosilane and dimethyldichlorosilane; (5) a sample application area closed with the photoresist is recovered; (6) the chip and a metal chip adapter are matched and mounted, wherein the sample application holes are pretreated with a covering matrix solution in advance, and the matrix solution is selected from a matrix solution for protein mass spectrometry or selected from a matrix solution for nucleic acid mass spectrometry.

Description

technical field [0001] The invention relates to a dual-purpose chip for detecting protein and nucleic acid by time-of-flight mass spectrometry, which can detect protein and nucleic acid molecules by mass spectrometry, and belongs to the field of mass spectrometry detection. Background technique [0002] Proteomics is a new science in the era of functional genomics, which studies the composition, activity rules and protein-protein interactions of intracellular proteins from an overall level. Including the identification of protein expression, modified form, structure, function and interaction. According to the research purpose, proteomics can be divided into expression proteomics, structural proteomics and functional proteomics. Expression proteomics is used to quantitatively study the expression of protein samples in cells. The purpose of proteomics is to map out the structure of protein complexes or proteins present in a special organelle, called structural proteomics, wh...

AI Technical Summary

Problems solved by technology

[0017] Although MALDI-TOF time-of-flight mass spectrometry can be used to detect proteins or nucleic acid molecules, from the application point of view, the traditional MALDI chip has a single application field, such as the DE1580TA steel target plate produced by SHIMADZU, which is only used for the identification of protein microorganisms and is suitable for The MALDI-TOF AXIMA of SHIMADZU; the L24 SpectroCHIP chip produced by AGENA is only used for genetic testing and is suitable for AGENA's MALDI-TOF MASSARRY. At present, there is no chip that can cover both genomics testing and proteomics identification.

[0018] From the perspective of the material and surface characteristics of the mass spectrometer chip, the traditional metal chip is repeatedly used, has scratches, and the surface is not chemically modified, the crystal form is poor, the accuracy of the mass spectrometer detection sample peak is low, the signal-to-noise ratio is low, and the baseline high

And for nucleic acid samples, small nucleic acid residues will affect mass spectrometry results

Therefore, metal chips are mainly used for protein spectrum detection, and cannot detect nucleic acid samples across borders.

Nucleic acid chips are mainly made of silicon wafers, but they are expensive, and the matrix formula is monopolized by foreign countries, and the surface of the purchased imported chips has been directly covered with the matrix, which limits their use in protein profiling.

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