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
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Embodiment 1
[0048] Example 1 Fluorescein isothiocyanate (FITC) labeled bovine serum albumin (BSA)
[0049] (1) Fabrication of micro-nano fluidic chips. The overall configuration of the micro-nano fluidic chip is as figure 1 As shown, it includes the preparation of single nanotubes; the preparation of microtubes; the reversible bonding of micro and nanotubes. The production of single nanotubes adopts the ultraviolet peeling 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 the nanotube is controlled by the time and intensity of ultraviolet light; the PDMS microtube manufacturing method uses the template method (see: C. Wang, J. Ouyang, HL Gao, HW Chen, JJ Xu, XH Xia, HY 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 nanochannels are reversibly bonded t...
Embodiment 2
[0051] Example 2 Fluorescein isothiocyanate (FITC) labeled immunoglobulin IgG
[0052] The preparation method of this embodiment is the same as that of embodiment 1, wherein in step 2, the protein is changed from bovine serum albumin to immunoglobulin IgG, and under other conditions unchanged, the fluorescently labeled product FITC-IgG is also obtained. Such as Picture 9 , Picture 9 It is the FITC fluorescence labeling spectrum of immunoglobulin on the micro-nano fluid control chip. From bottom to top, it is the purified products FITC-IgG, FITC, and unpurified reaction products (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 embodiment is the same as that of embodiment 1, wherein the fluorescent probe in step 2 is replaced with positively charged rhodamine isothiocyanate (RBITC), and under other conditions unchanged, the fluorescent labeled product RBITC-BSA is also obtained. Such as Picture 10 , Picture 10 It is the fluorescence photo of bovine serum albumin labeled with rhodamine isothiocyanate on the micro-nano fluid control chip. Picture a is the photo before purification, and b is the picture after purification. It can be seen that the micro-nano fluid control chip has an effect on bovine serum albumin. (BSA) marking effect is better.
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