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Micro-nanofluidic chip and method for achieving rapid fluorescent labeling of proteins

A fluorescent labeling and micro-nanofluidic technology, applied in the field of protein, can solve problems such as complex structure and additional purification process, and achieve the effect of simple operation, high-efficiency enrichment ability, and accelerated speed

Active Publication Date: 2014-06-18
NANJING UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The purpose of the present invention is to provide a micro-nanofluidic chip for fast fluorescent labeling of proteins, which solves the problem that the current fluorescent labeling of proteins requires an additional purification process of the labeled product and has a complex structure

Method used

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  • Micro-nanofluidic chip and method for achieving rapid fluorescent labeling of proteins
  • Micro-nanofluidic chip and method for achieving rapid fluorescent labeling of proteins
  • Micro-nanofluidic chip and method for achieving rapid fluorescent labeling of proteins

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0048] Example 1 Fluorescein isothiocyanate (FITC)-labeled bovine serum albumin (BSA)

[0049] (1) Fabrication of micro-nanofluidic chips. The overall configuration of the micro-nanofluidic chip is as follows: figure 1 As shown, including the preparation of single nanopipes; the preparation of micropipes; the reversible bonding of micropipes and nanopipes. The fabrication of single nanopipes adopts the ultraviolet stripping method (see: C. Wang, J. Ouyang, H. L. Gao, H. W. Chen, J. J. Xu, X. H. Xia, H. Y. Chen, Talanta , 2011, 85, 298-303), the depth of nanotubes is regulated by the time and intensity of ultraviolet light; the template method is used for the fabrication of PDMS microchannels (see: C. Wang, J. Ouyang, H. L. Gao, H. W. Chen, J. J. Xu, X. H. Xia, H. Y. Chen, Talanta , 2011, 85, 298-303; and C. Wang, S. J. Li, Z. Q. Wu, J. J. Xu, H. Y. Chen, X. H. Xia, Lab Chip , 2010, 10, 639-646); Finally, the PDMS cover sheet with microchannels and the substrate with...

Embodiment 2

[0051] Example 2 Fluorescein isothiocyanate (FITC)-labeled immunoglobulin IgG

[0052]The preparation method of this example is the same as that of Example 1, wherein the protein in step 2 is replaced by immunoglobulin IgG from bovine serum albumin, and the fluorescently labeled product FITC-IgG is also obtained under other conditions unchanged. Such as Figure 9 , Figure 9 It is the FITC fluorescent labeling spectrogram of immunoglobulin on the micro-nanofluidic chip. From bottom to top, it is the purified product FITC-IgG, FITC, and unpurified reaction product (including FITC-IgG and unreacted FITC). It can be seen that this method of labeling proteins on micro / nanochips can be successfully used to label other proteins.

Embodiment 3

[0053] Example 3 Rhodamine isothiocyanate (RBITC) labeled bovine serum albumin (BSA)

[0054] The preparation method of this example is the same as that of Example 1, wherein the fluorescent probe in step 2 is replaced with positively charged rhodamine isothiocyanate (RBITC), and the fluorescently labeled product RBITC-BSA is also obtained under other conditions unchanged. Such as Figure 10 , Figure 10 It is the fluorescent photo of bovine serum albumin labeled with rhodamine isothiocyanate on the micro-nanofluidic chip, a is the photo before purification, and b is the photo after purification. It can be seen that the micro-nanofluidic chip has no effect on bovine serum albumin (BSA) marked better.

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Abstract

The invention relates to a micro-nanofluidic chip for achieving rapid fluorescent labeling of proteins, which is designed according to different sizes of the proteins and a fluorescent probe. A nano-structure is led in a micro-nanofluidic system to construct a micro-nanofluidic chip system. The proteins are gathered through a nano channel, and the labeled proteins are gathered through a micro-channel. The invention further provides a method for achieving teh rapid fluorescent labeling of the proteins by using the micro-nanofluidic chip. The micro-nanofluidic chip and the method for achieving the rapid fluorescent labeling of proteins can achieve the rapid fluorescent labeling of the proteins, are particularly suitable for microminiaturization labeling of microgram-level and nanogram-level proteins, nucleic acid and other precious bio-molecules, do not need additional purification processes and chromatographic separation to remove excess unreacted dye molecules, and are simple and convenient to operate.

Description

technical field [0001] The invention belongs to the field of proteins, and relates to a micro-nano fluidic chip and a method for realizing rapid fluorescent labeling of proteins, in particular to a micro-nano fluidic chip and a method thereof that do not require an additional purification process of labeled products and have a simple structure. Background technique [0002] Fluorescence detection is extremely sensitive and can achieve single-molecule level analysis, so it is widely used in life sciences, environmental sciences, medicine, pharmacy, and in exploring the intrinsic functions of biomolecules, cells, and organisms. However, only a small number of biomolecules in nature have fluorescence properties, and most biomolecules cannot be directly detected by fluorescence analysis. Therefore, only after the biomolecules (antibodies, proteins, amino acids, polypeptides) are labeled with fluorescent dyes can ultra-sensitive analysis and detection of biomolecules be achieved...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/68G01N21/64B01L3/00
Inventor 夏兴华王琛
Owner NANJING UNIV
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