Rapid protein analysis and detection device based on whole microfluidic chip closing system
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A microfluidic chip and rapid analysis technology, applied to the analysis of materials, instruments, etc., can solve the problems of complex operation and poor accuracy, and achieve the effects of high sensitivity, high detection, and reduced instrument volume
Active Publication Date: 2015-03-18
DALIAN INST OF CHEM PHYSICS CHINESE ACAD OF SCI
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At present, commonly used detection methods such as chemical test paper method, ELISA, SDS-PAGE and other methods have problems such as poor ac
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[0025] Example 1
[0026] Such as figure 1 Construct a rapid protein analysis and detection device based on a microfluidic chip, which integrates a microfluidic control system, a sampling system, a microfluidic chip system, a miniature laser induced fluorescence detection system, and a data acquisition system. The size of the entire instrument is 30cm×15cm× 50cm.
[0027] Such as figure 2 As shown, a microfluidic chip for protein enrichment-elution-on-line fluorescent labeling-detection is constructed. Among them, the enrichment column is an albumin antibody column, and the particles in the antibody column are Agilent's packing material that supports albumin antibody. The fluorescent labeling reagent is 2% Nano Orange, and 0.5M NaHCO is used. 3 As a derivatization buffer solution. Buffer A and Buffer B are commercial loading and elution solutions of Agilent. The excitation / emission wavelength of laser-induced fluorescence detection is 473 / 570nm.
[0028] The specific operation pr...
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[0033] Example 2
[0034] The constructed rapid protein analysis and detection device based on microfluidic chip is used to quantitatively analyze the urine samples of patients.
[0035] Use 0.01mg / mL, 0.03mg / mL, 0.05mg / mL, 0.07mg / mL, 0.1mg / mL as standard samples for analysis, calculate the detected peak area, and make a standard curve (such as Figure 4 As shown), the linear relationship between concentration (x) and peak area (y) is y=57.454x-0.5632, R 2 =0.9625.
[0036] Take the patient’s urine sample by centrifugation at 3000g at 4℃ for 5min, filter with 0.22um membrane, and precipitate with an equal volume of acetone, then centrifuge at 12000g at 4℃ for 5min, remove the supernatant, and redissolve the precipitate 1mL buffer A (Agilent), diluted 500 times, for analysis (such as Figure 5 Shown). The peak area is 0.6518. According to the linear relationship between concentration (x) and peak area (y), y=57.454x-0.5632, the calculated concentration is 0.2115 mg / mL, and the album...
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Abstract
The invention relates to a micro device for performing on-line enrichment, elution, fluorescence labeling and detection on protein, and in particular provides a rapid protein analysis and detection device based on whole microfluidic chip closing system. The device is composed of a micro fluidic controlling system, a miniature laser induced fluorescence detection system, a microfluidic chip, an injection valve, a connecting pipeline, an interface and a data acquisition and processing part. A protein sample is enriched by an enriching column in a chip passage, is subjected to on-line fluorescence labeling operation and elution operation at the same time, and is induced by laser to be subjected to fluorescence detection. The rapid protein analysis and detection device has the advantages that the sample pre-processing process, fluorescence labeling and detection are integrated so as to enable cumbersome steps of an off-line analysis method to be omitted, the sample processing time is greatly shortened and the high sensitivity detection is achieved; the whole device is small in size and low in energy consumption; the microfluidic chip and a micro injection pump are connected through the interface with a special design; a whole fluid system runs in the closing system, the detection requirement under a special condition (such as non-gravity state) can be met. Thereby, the rapid protein analysis and detection device has the very good practical application value.
Description
technical field [0001] The invention relates to a protein detection device, and in particular provides a rapid protein analysis and detection device based on a microfluidic chip. Background technique [0002] Micro Total Analysis Systems (μ-TAS) is a new interdisciplinary field, the ultimate goal of which is to maximize the transfer of analytical laboratory functions to portable analytical instruments through the miniaturization and integration of chemical equipment. In the equipment (such as various types of chips), the "miniaturization" of the analysis equipment is realized. Its characteristic is that it can realize the sampling, dilution, addition of reagents, enrichment, extraction, mixing, reaction, separation, detection, etc. of the target analyte. Integration of steps. [0003] There are trace amounts of protein in body fluid, which is one of the important indicators to reveal the health of the human body, and is very important for evaluating its health status. Curr...
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IPC IPC(8): G01N35/00
Inventor 张丽华戴忠鹏梁玉杨开广张玉奎
Owner DALIAN INST OF CHEM PHYSICS CHINESE ACAD OF SCI
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