Carrier for in-situ analysis of intracellular protein compound as well as preparation method and application of carrier

A technology for in situ analysis of protein complexes, applied in the field of carrier for in situ analysis of intracellular protein complexes and its preparation and application, can solve the difficulties of multiple side reactions and in situ targeted analysis of intracellular protein complexes , strong activity of cross-linking agent and other issues, to achieve low cytotoxicity, solve the difficulty of in situ targeted analysis, and solve the effect of strong activity

Active Publication Date: 2020-12-15
DALIAN INST OF CHEM PHYSICS CHINESE ACAD OF SCI
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  • Abstract
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Problems solved by technology

[0008] The purpose of the present invention is to provide a carrier for in situ analysis of intracellular protein complexes and its preparation method and application, through the carrier of the present invention for in sit

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  • Carrier for in-situ analysis of intracellular protein compound as well as preparation method and application of carrier

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Example Embodiment

[0028] Example 1

[0029] 2 parts by mass of castor oil polyoxyethylene ether, 15 parts by mass of poly (lactide-glycolide) with molecular weight of 10,000, 1 part by mass of didodecyldimethylammonium bromide and 4 parts by mass of cross-linking agent (bissuccinimide octanoate) were dissolved in 500 parts by mass of dichloromethane. The above mixture was poured into 1% polyvinyl alcohol aqueous solution, and the carrier was cured with 1% polyvinyl alcohol aqueous solution after ultrasonic treatment for 2 minutes. TEM photos of the carrier prepared in this example are as follows Figure 1 Shown. The particle size and surface potential of the prepared materials are shown in Table 1.

[0030] Table 1.DLS test data (1, 2, 3 are three DLS tests of carrier)

[0031] 1 2 3 Particle size / nm 274.6 268.8 273.7 PDI 0.237 0.181 0.135 Zeta potential / mv 38 39.9 41

[0032] by Figure 1 TEM characterization shows that the synthesized empty PLGA nanoparticles...

Example Embodiment

[0034] Example 2

[0035] 3 parts by mass of castor oil polyoxyethylene ether, 15 parts by mass of polylactide-glycolide with molecular weight of 20,000, 1 part by mass of didodecyldimethylammonium bromide, and 5 parts by mass of cross-linking agent (bissuccinimide octanoate) were dissolved in 400 parts by mass of dichloromethane. The above mixture was poured into 1% aqueous solution of polyvinyl alcohol, and the carrier was further cured with 1% aqueous solution of polyvinyl alcohol. The particle size of the prepared carrier is 126.6nm±0.75nm, and the potential (Z-potential: 46.6 0.379 mV) is in line with the conditions of its transmembrane transport carrier.

[0036] In the research of cross-linking agent carrier used for in-situ analysis of protein complex, the carrier and cells were co-incubated in 1640 culture medium, and after incubation for 3 hours, the cells were collected. Then, protein and protein complex was extracted from the cells by ionic liquid, and the extracted pr...

Example Embodiment

[0037] Example 3

[0038] 5 parts by mass of castor oil polyoxyethylene ether, 15 parts by mass of polylactide-glycolide with molecular weight of 80,000, 2 parts by mass of didodecyldimethylammonium bromide, and 8 parts by mass of cross-linking agent (bissuccinimide octanoate) were dissolved in 700 parts by mass of dichloromethane. The above mixture was poured into 0.5% aqueous solution of polyvinyl alcohol, and the carrier was further cured with 2% aqueous solution of polyvinyl alcohol. The particle size of the prepared carrier is 236.6nm±0.92nm, and the potential (Z-potential: 49 0.379 mV) is in line with the conditions of its transmembrane transport carrier.

[0039] In the research of cross-linking agent carrier used for in-situ analysis of protein complex, the carrier and cells were co-incubated in 1640 culture medium, and after incubation for 6 hours, the cells were collected. Then, protein and protein complex was extracted from the cells by ionic liquid, and the extracted p...

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Abstract

The invention relates to a carrier for in-situ analysis of an intracellular protein compound as well as a preparation method and application of the carrier. The carrier for in-situ analysis of the intracellular protein complex is used for conveying a chemical cross-linking agent for protein complex analysis to a specific part in a cell in a transmembrane delivery manner and releasing the chemicalcross-linking agent, so that the protein complex is subjected to in-situ cross-linking. And analyzing the spatial-temporal dynamic change rule of the intracellular protein compound by combining a massspectrometry technology. The method has great significance in understanding the life process, revealing the disease mechanism, screening biomarkers and searching drug targets.

Description

technical field [0001] The invention relates to a carrier for in-situ analysis of intracellular protein complexes, a preparation method and application thereof. The invention belongs to functional biological material and its preparation and application in in situ analysis of protein complex. Background technique [0002] Protein complexes are important building blocks of protein machinery. High-dimensional signal transduction networks dynamically assembled by protein complexes run through different subcellular organelles and regulate almost all life processes. Analyzing the temporal and spatial dynamic changes of protein complexes in complex systems and drawing interaction networks are of great significance for understanding life processes, revealing disease mechanisms, screening biomarkers, and finding drug targets. [0003] Chemical cross-linking combined with mass spectrometry (referred to as cross-linking mass spectrometry or CXMS) is a new method developed in recent y...

Claims

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Application Information

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IPC IPC(8): G01N30/02G01N30/06
CPCG01N30/02G01N30/06G01N2030/067
Inventor 张丽华陈玉宛杨开广周雯裴家琪梁振张玉奎
Owner DALIAN INST OF CHEM PHYSICS CHINESE ACAD OF SCI
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