Method for modifying titanium dioxide nanofilm on target plate and application thereof

A nano-film, titanium dioxide technology, applied in the field of MALDI target plate, can solve the problems of inability to guarantee consistency, limited firmness of enrichment material and target plate, ion source pollution, etc., to achieve simple and time-saving enrichment process and long storage time. , good repeatability

Inactive Publication Date: 2017-06-23
FUDAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

The disadvantage of these methods is that the consistency of the thickness of titanium dioxide at each point cannot be guaranteed, that is, the repeatability between points, and the degree of firmness of the enrichment material and th

Method used

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  • Method for modifying titanium dioxide nanofilm on target plate and application thereof
  • Method for modifying titanium dioxide nanofilm on target plate and application thereof
  • Method for modifying titanium dioxide nanofilm on target plate and application thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0029] Example 1: Preparation of a target plate for modifying titanium dioxide nano film.

[0030] (1) Wipe the MALDI target plate to be modified with a dust-free paper dipped in an appropriate amount of detergent, and then ultrasonically clean it with 1% formic acid solution, ethanol solution and deionized water for 10 minutes each, and then clean it again with deionized water. Rinse and dry;

[0031] (2) Put the target plate obtained in step (1) into an atomic layer deposition system, using tetrakis(dimethylamino)titanium (TDMAT) and deionized water as titanium source and oxidation source, respectively. In the reaction chamber, the MALDI target plate was alternately exposed to TDMAT and deionized water, and high-purity argon was used as the carrier gas and purge gas. For all samples, one cycle during deposition consisted of a 0.2 s TDMAT pulse, a 10 s rinse, a 0.2 s deionized water pulse, and a 10 s rinse. The desired film thickness is achieved by varying the number of cyc...

Embodiment 2

[0034] Example 2: Application of titanium dioxide modified target plate on the target to purify phosphorylated peptides in β-casein enzymatic hydrolysis solution.

[0035] (1) Preparation of standard protein enzymatic hydrolysis solution: Accurately weigh standard protein β-casein and BSA and dissolve them in 25 mM ammonium bicarbonate buffer solution respectively, so that the protein concentration is 1 mg / mL, boil for 10 minutes, after cooling, follow the enzymatic Add an appropriate amount of trypsin (trypsin) at a ratio of 1:40 to the protein, and digest at 37°C for 16 hours.

[0036] (2) Purification of phosphorylated peptides on the target and MALDI analysis: Rinse and dip the modified target plate with enrichment buffer (50% acetonitrile aqueous solution containing 0.1% trifluoroacetic acid) for 1 minute for activation and then dry it. Then dilute the β-casein hydrolysis solution with enrichment buffer to an appropriate concentration, and pipette 0.8 μL onto the modified...

Embodiment 3

[0038] Example 3: Investigation of the selectivity of purified phosphorylated peptides on the target.

[0039] (1) Preparation of mixed protein enzymolysis solution: prepare a mixture of β-casein and BSA enzymolysis solution with a molar ratio of 1:200, dilute to an appropriate concentration with enrichment buffer (make the concentration of β-casein 400 fmol / uL ), used to investigate the selectivity of purified phosphorylated peptides from the target plate.

[0040] (2) Investigation of the selectivity of purified phosphorylated peptides on the target: Rinse and dip the modified target plate with enrichment buffer (50% acetonitrile aqueous solution containing 0.1% trifluoroacetic acid) for 1 minute for activation and then dry it. Use a pipette gun to pipette 1 μL of the mixture onto the modified target plate, and then place the target plate in a wet box with a lid for static incubation for about 10-30 minutes. Afterwards, the target plate was quickly and fully washed several ...

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Abstract

The invention belongs to the field of advanced nanomaterials and nanotechnology, and specifically provides a preparing method for modifying a titanium dioxide nanofilm on a target and application thereof. The method applies an atomic layer deposition technology, a layer of titanium dioxide nanofilm is modified on a stainless steel MALDI target plate, and the layer of film and the target plate are combined very firmly. Afterwards, the modified target plate is applied to purification and MALDI analysis of phosphorylated peptides on a target. The target plate in the invention displays excellent purification performance, has well repeatability and has broad application prospect in the field of purification of the phosphorylated peptides.

Description

technical field [0001] The invention belongs to the field of advanced nanomaterials and nanotechnology, and in particular relates to a method for preparing a MALDI target plate modified with a titanium dioxide film and an application thereof for enriching phosphorylated peptides. Background technique [0002] Protein phosphorylation is one of the most important protein post-translational modifications, involved in many cellular life processes, such as cell proliferation, differentiation, apoptosis and other cellular activities. Since abnormal phosphorylation will lead to various diseases and cancers, phosphoproteomics of living systems has been extensively and intensively studied in order to provide more information for the clinicopathology and diagnosis of these diseases. Nowadays, due to the high-throughput and rapidity of mass spectrometry detection, most phosphoproteomics research is based on mass spectrometry strategies, especially MALDI-TOF mass spectrometry (matrix-as...

Claims

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Application Information

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IPC IPC(8): C23C16/40C23C16/455
CPCC23C16/405C23C16/45534
Inventor 邓春晖林海珠袁凯平
Owner FUDAN UNIV
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