A kind of zip-tips for capturing thiol-containing peptides and its preparation method

A peptide and sulfhydryl technology, applied in the field of Zip-Tips (separation tips) and its preparation, can solve the problems of long time, separation speed and separation efficiency that do not meet the needs of rapid development, multiple experimental steps, etc., and achieve a wide range of applications. The prospect, the production method is simple and novel, and the cost is low

Inactive Publication Date: 2015-07-29
FUDAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the current limited separation speed and separation efficiency do not meet the needs of rapid development in the future, so there is an urgent need to develop a simpler and more efficient separation technology
For example, using related chromatographic techniques, sulfhydryl-containing peptides can be captured by disulfide bond exchange or derivatization methods, but these methods generally require many experimental steps and take a long time

Method used

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  • A kind of zip-tips for capturing thiol-containing peptides and its preparation method
  • A kind of zip-tips for capturing thiol-containing peptides and its preparation method
  • A kind of zip-tips for capturing thiol-containing peptides and its preparation method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] Example 1. Preparation of novel Zip-Tips for capturing sulfhydryl peptides.

[0021] (1) Pretreatment of the tip: Heat the tweezers with a candle, and then clamp the end of the ordinary tip.

[0022] (2) Preparation of gold modified polymer: Dissolve 10.02 mg epoxy-based material and 1.59 mg glutathione in 1 mL pH=9-10 borax buffer solution, shake at 37 ℃ for 24 hours, and then centrifuge to obtain The precipitate was reacted with the synthesized 1 mL gold gel at 37°C for another 1 hour, and centrifuged.

[0023] (3) Preparation of the new Zip-Tips: The prepared gold-modified polymer is filled into the pretreated pipette tip by pressure with a pipette. At this time, the liquid flows out, and the filler remains in the pipette tip due to the large particle size .

Embodiment 2

[0024] Example 2. Zip-Tips are used to capture peptides containing sulfhydryl groups and combine with MALDI-TOF-MS for efficient and rapid analysis.

[0025] (1) Sample injection: Use a pipette gun to inject the sample from the upper end of the Zip-Tips, and apply pressure through the pipette gun to make the sample slowly flow through the packing.

[0026] (2) Elution: use NH first 4 HCO 3 (10mM)+5% acetonitrile (volume ratio) to wash away the weakly bound interfering substances, and then use 70% acetonitrile and 0.1% TFA (volume ratio) to elute the peptides containing sulfhydryl groups from the Zip-Tips.

[0027] (3) Point target: Pipette 1uL analyte on the MALDI target plate and place it in the air to dry naturally at room temperature; drop 0.5 uL CHCA solution on the analyte layer and place it in the air to dry naturally at room temperature.

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Abstract

The invention belongs to the technical field of isolation analysis, and particularly relates to a Zip-Tips (isolation gun) for trapping a sulfhydryl-containing peptide segment and a preparation method thereof. The method provided by the invention comprises the following steps: oscillating a polymer material containing epoxide group and glutathione in an alkaline buffer solution, centrifuging, carrying out oscillating reaction on the precipitate and synthesized gold size, centrifuging to obtain a gold-modified polymer material, and finally, filling the gold-modified polymer material in the gun; and flattening the tail end of the gun with heated tweezers while ensuring that only liquid flows out but the filler can not spill over, thereby obtaining the novel Zip-Tips for sensitively trapping a sulfhydryl peptide segment. The method provided by the invention has the advantages of simple operation steps and very low cost, can be used repeatedly, and can be used for quickly and sensitively analyzing the sulfhydryl-containing peptide segment when being in combination with high-flux MALDI-TOF-MS (matrix-assisted laser desorption / ionization time-of-flight mass spectrometry).

Description

Technical field [0001] The invention belongs to the technical field of separation and analysis, and specifically relates to a Zip-Tips (separation tip) used in the pretreatment step of proteomics samples and a preparation method thereof. Background technique [0002] In the fields of biochemistry and biomedicine, mass spectrometry has helped discover and detect many important peptides. Peptides and materials are captured and separated by a specific force, and then mass spectrometry is used for rapid and efficient analysis. Among them, MALDI-TOF-MS (Matrix-Assisted Laser Desorption / Ionization Time-of-Flight Mass Spectrometry) has become a very popular analytical technique due to its advantages of rapidity, sensitivity, and high throughput. But before using MALDI-TOF-MS to detect the peptide mixture, the peptide mixture needs to be separated and enriched for better analysis. This means that separation and enrichment methods play an important role in MALDI-TOF-MS analysis. However...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N1/28G01N1/34
Inventor 高明霞张祥民万丹陈奇
Owner FUDAN UNIV
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