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Micro-cavity array mass spectrum target plate as well as manufacturing method and application thereof

A microcavity and array technology, applied in screening applications, in the field of microcavity array mass spectrometry target plates, can solve the problems of limited screening capacity, low screening efficiency, and high cost, and achieve the effect of high-throughput screening and detection

Active Publication Date: 2014-06-18
THE NAT CENT FOR NANOSCI & TECH NCNST OF CHINA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are still deficiencies in these microarray chip platforms, mainly in that the throughput of peptide library screening based on microarray chips generally does not exceed 10 4 , the screening capacity is limited; the microarray peptide library preparation and screening process requires special reagents such as electrosensitivity and light sensitivity, and extremely complex software and hardware platforms, which are expensive
However, the current microfluidic chip can only complete the high-throughput peptide library screening itself online, while the more important sequencing and identification must be completed offline, and the screening efficiency is low

Method used

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  • Micro-cavity array mass spectrum target plate as well as manufacturing method and application thereof
  • Micro-cavity array mass spectrum target plate as well as manufacturing method and application thereof
  • Micro-cavity array mass spectrum target plate as well as manufacturing method and application thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0052] Example 1: Fabrication of Microcavity Array Mass Spectrometry Target Plate

[0053] Such as figure 1 As shown, the microcavity array mass spectrometer target plate 1 provided in this embodiment includes a substrate 2, an adhesive layer 3 formed on the substrate 2, and a conductive layer 4 formed on the adhesive layer 3, so The side of the microcavity array mass spectrometry target plate 1 with the conductive layer 4 is provided with a microcavity area ( figure 1 The area where the array is located), the microcavity 5 is etched on the substrate 2 at the position of the microcavity area, and the microcavity 5 is opened in the conductive layer 4 and arranged in an array. For calibration needs, a standard sample calibration microcavity 6 may also be included.

[0054] Such as figure 2 As shown, the microcavity 5 is designed as a cube (the top view of the microcavity 5 is shown in the figure), the side length of which is 250 μm, and the distance between adjacent microcav...

Embodiment 2

[0069] Example 2: Using the microcavity array mass spectrometry target plate made in Example 1 to screen the polypeptide library

[0070] (1) Construction of peptide library

[0071] The nonapeptide fragment HA of the hemagglutinin protein was selected as the model polypeptide (sequence YPYDVPDYA). The HA antibody AHA was selected as the receptor protein that could specifically bind to it, and a model octapeptide library containing HA was established. The peptide library construction method uses amino-modified TentaGel (swellable particle size can be 200 microns) resin as a solid phase carrier, and before the first amino acid is synthesized, methionine is first coupled at the C-terminal. The Fmoc synthesis strategy was used for mixed and evenly divided synthesis, and the four amino acids Y, P, D, and V constituting the HA nonapeptide were selected as reaction units. After six rounds of synthesis, the capacity of the library was constructed to be 4 6 The peptide library, the ...

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Abstract

The invention relates to a micro-cavity array mass spectrum target plate as well as a manufacturing method and application thereof. The micro-cavity array mass spectrum target plate comprises a substrate, an adhesion layer formed on the substrate and a conducting layer formed on the adhesion layer, wherein the side with the conducting layer of the micro-cavity array mass spectrum target plate is provided with a micro-cavity region consistent with a surface plasma resonance imaging detection region; micro-cavities are etched in the substrate in the position of the micro-cavity region, are opened on the conducting layer and are arranged into an array. The micro-cavity array mass spectrum target plate provided by the invention can be used for screening and enriching anode targets in a one-bead one-compound peptide library and directly carrying out in-situ substrate-assisted laser desorption ionization and flight time mass spectrometric detection on the anode targets; in addition, the micro-cavity array mass spectrum target plate can be combined with surface plasma resonance imaging to carry out in-situ affinity identification, so that the efficient and high-throughput screening and detection on the peptide library can be realized.

Description

technical field [0001] The invention relates to the technical field of microarray chips, in particular to a microcavity array mass spectrometry target plate, a manufacturing method thereof and an application in screening of a combinatorial chemical polypeptide library. Background technique [0002] Lab on a chip has developed into one of the most cutting-edge technological fields in the world today (Sens.Actutors, B, 1990, 1, 244-248.). Microchip technology integrates or basically integrates the basic operation units involved in the fields of chemistry and biology, such as pretreatment, sample addition, reaction, separation, and analysis, etc. functional technology platform. [0003] There are two methods for the synthesis of combinatorial compound libraries: liquid-phase combinatorial synthesis and solid-phase combinatorial synthesis. Since the products obtained by solid-phase synthesis are easier to purify, excess reagents can be used, and automation is easier, it has be...

Claims

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Application Information

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IPC IPC(8): G01N27/64G01N21/552
Inventor 王蔚芝李梦林魏泽文胡志远
Owner THE NAT CENT FOR NANOSCI & TECH NCNST OF CHINA
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