Functional protein and cyanine dye molecule compound and preparation method and application thereof

A cyanine dye, protein technology, applied in material excitation analysis, preparations for in vivo experiments, material analysis by optical means, etc., can solve the problems of easy diffusion in vivo, poor in vivo stability, short imaging window, etc. Signal-to-noise ratio and penetration depth, slower dispersion, and less background scattering

Active Publication Date: 2019-09-06
SHANGHAI THERANOSTICS BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, cyanine dyes have relatively low luminous efficiency in the second near-infrared region, are prone to photobleaching, have poor stability in vivo, and are easily diffused in vivo, resulting in a short imaging window.

Method used

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  • Functional protein and cyanine dye molecule compound and preparation method and application thereof
  • Functional protein and cyanine dye molecule compound and preparation method and application thereof
  • Functional protein and cyanine dye molecule compound and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0058] Select three typical cyanine dyes ICG, IR-783 and IR-12N3 respectively figure 1 The 4 schemes provided are coated with albumin, and the specific schemes are as follows:

[0059] Scheme 1: Add 500 microliters of PBS to 500 microliters of albumin solution, add 11 microliters of cyanine dye, mix well, react in a shaking box at 37 degrees Celsius for 24 hours, wash five times with a 30k centrifugal filter membrane, and post-process The temperature is 60 degrees Celsius for 10 minutes.

[0060] Scheme 2: Add 500 microliters of PBS to 500 microliters of albumin solution, add 60 microliters of glutathione solution and mix well; add 11 microliters of cyanine dye and mix well, react in a shaking box at 37 degrees Celsius for 24 hours, use 30k The centrifugal filter membrane was washed five times, and the post-treatment temperature was 60 degrees Celsius for 10 minutes.

[0061] Scheme 3: Add 500 microliters of PBS to 500 microliters of albumin solution, add 60 microliters of ...

Embodiment 2

[0065] Example 2. Preparation of complexes of IR-783 and albumin by simple blending

[0066] Preparation scheme such as figure 1 As shown in "Scheme 1", it specifically includes: adding IR-783 solution to 40mg / ml (602μM) BSA solution, controlling the molar ratio of albumin and IR-783 to 1:1, mixing, and controlling albumin The reaction concentration is 20 mg / ml, reacted in a shaking box at 37 degrees Celsius for 24 hours, washed five times with a 30k centrifugal filter membrane, and the post-treatment temperature was 60 degrees Celsius for 10 minutes. The reaction process and the obtained complex structure can be found in figure 1 , 2 Schematic of , nanoparticle-like under the transmission electron microscope, see Figure 8 Shown in the leftmost figure. The resulting complex was designated as "IR-783@BSA".

Embodiment 3

[0067] Example 3. Preparation of complexes of IR-783 and albumin by reduction of glutathione after blending

[0068] Preparation scheme such as figure 1 As shown in "Scheme 2", it specifically includes: adding 60 microliters of 0.25M glutathione solution containing 15% DMSO to 40mg / ml (602μM) BSA solution, and then adding IR-783 solution to control the interaction between albumin and IR-783 reaches a molar feed ratio of 1:1, mixes well; and controls the albumin reaction concentration at 20mg / ml, reacts in a shaking box at 37 degrees Celsius for 24 hours, washes five times with a 30k centrifugal filter membrane, and the post-treatment temperature is 60 degrees Celsius for 10 minutes, the reaction process and the structure of the resulting complex can be found in figure 1 , 2 Schematic of , nanoparticle-like under the transmission electron microscope, see Figure 8 Shown in the second picture from the left. The resulting complex was designated as "IR-783@BSA-GSH".

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Abstract

The invention provides a functional protein and cyanine dye molecule compound and further provides a preparation method of the compound and application of the compound as a fluorescence molecular imaging developer in a second near-infrared region. The functional protein and cyanine dye molecule compound is a compound formed by controllable self-assembly of cyanine dye molecules and functional protein. The outer layer of the compound is provided with the functional protein which wraps the cyanine dye molecules internally. Compared with an existing second near-infrared region probe, the compoundhas advantages that luminescence and in-vivo behaviors are evidently improved, imaging effects in the second near-infrared region are remarkably improved, and the compound can be applied to in-vivo fluorescence imaging and molecular imaging.

Description

technical field [0001] The invention belongs to the technical field of near-infrared imaging probes, and specifically relates to a novel compound based on functional proteins and cyanine dyes, a preparation method thereof, and an application as a near-infrared region two complex probe. Background technique [0002] At present, the development of biological imaging technology has greatly enriched the means for researchers to explore biological processes and mechanisms. Fluorescence imaging has the advantages of fast imaging speed, high sensitivity, and targeted design, and is widely used in cells and tissues. and in vivo imaging. Biological imaging mainly uses specific fluorescent probes and selects appropriate imaging equipment (including wide-field or microscopic equipment sensitive from the visible region to the near-infrared region) to study biological processes and information. The key to fluorescence imaging technology is to use sufficient signal contrast to distinguis...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K49/00G01N21/64
CPCA61K49/0034A61K49/0032A61K49/0056A61K49/0058A61K49/0093G01N21/6486G01N21/6456
Inventor 陈小元田蕊朱守俊
Owner SHANGHAI THERANOSTICS BIOTECH CO LTD
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