Simple and rapid protein detection device and method

Abstract

To provide a simple and rapid protein detection device.SOLUTION: A western blotting method used in conventional protein detection requires several days to obtain results and needs a special device such as an electrophoresis device. In addition, a mass spectrometry method and a flow cytometry method need expensive instruments. The present invention provides a device capable of simply and rapidly detecting and quantifying specific proteins, and includes the steps of: reacting proteins in a sample with antibodies; passing the protein-antibody complex between two plastic plates or glass plates; capturing specific proteins; and detecting the specific proteins.SELECTED DRAWING: Figure 1

Description

Technical Field

[0001] The present invention relates to an apparatus and a method capable of detecting and quantifying a specific protein.

Background Art

[0002] In the research of medical biology and pharmacy, the Western blotting method has been conventionally used for protein detection. This is a method in which a protein suspension is electrophoresed and then transferred to a membrane, and the protein adsorbed on the membrane is detected by an antibody labeled with a label.

Prior Art Documents

Patent Documents

[0003]

Patent Document 1

Non-Patent Documents

[0004]

Non-Patent Document 1

Non-Patent Document 2

Summary of the Invention

Problems to be Solved by the Invention

[0005] The Western blotting method involves frequent operations and requires a period of 1 to 1.5 days to obtain results. Also, the ELISA method is a one-day operation and requires a dedicated plate. Other protein detection methods such as mass spectrometry and flow cytometry also exist, but they require very expensive machines. In the Western blotting method, since electrophoresis is used, low-molecular-weight and high-molecular-weight proteins outside the gel range cannot be detected, and due to the presence of contaminants with the same molecular weight, they may not be detected by antibodies. There is a need for a simpler, shorter-time, and lower-cost protein detection and analysis method.

Means for Solving the Problems

[0006] The present invention includes the following aspects. [1] A method of reacting a sample containing a protein to be detected with a specific antibody [2] A method of passing a protein-antibody complex between two plastic plates or glass plates [3] A method of capturing a protein-antibody complex [4] A method of detecting and quantifying the captured protein

[0007] The apparatus according to the present invention does not require large-scale equipment such as an electrophoresis apparatus, a membrane for transfer, a mass spectrometer, or a flow cytometer, and is simple and low-cost. Detection and quantification can be performed within a significantly shortened working time and detection time.

Effects of the Invention

[0008] The present invention can provide a technique that does not require large-scale equipment, is simple and low-cost, and can quickly detect and quantify a specific protein.

Brief Description of the Drawings

[0009]

Figure 1

Embodiments for Carrying Out the Invention

[0010] Figure 1 shows an embodiment of the protein detection apparatus according to the present invention. Two different labeled antibodies are reacted with the specific protein to be detected in the sample. One is a fluorescently labeled antibody and the other is a biotinylated antibody.

[0011] A sample containing a protein-antibody complex is administered between two plastic plates or glass plates and is moved between the plastic plates by capillary action. A capillary may be used instead of the two plates. Avidin is immobilized at the detection site. Biotin has a strong binding force with avidin, and the protein-antibody complex is captured in the avidin immobilized region of the plastic plate. When there is a protein that binds to the two antibodies, the fluorescently labeled antibody remains in the avidin immobilized region via the protein. This region is detected by a fluorescence plate reader.

[0012] Instead of a fluorescence plate reader, it can also be detected by an In cell analyzer or a fluorescence microscope. In addition to fluorescence, it is also possible to label the antibody with a coloring substance or an enzyme that causes color development / light emission. In that case, a detector that can detect color development / light emission is used.

[0013] At the same time, proteins that are always expressed, such as actin and GAPDH, are simultaneously detected with other fluorescently labeled antibodies and used as a loading control. By doing so, quantitative data corrected for differences between devices can be obtained.

[0014] By using antibodies for different types of proteins for detection, protein-protein interactions in the sample can be detected.

[0015] By using a protein / peptide / compound instead of the antibody used for detection, interactions with a specific protein can be detected.

[0016] According to the present invention, it is possible to replace the Western blotting method and ELISA method performed in the laboratory and detect proteins more simply, quickly, and at low cost. It can also be used for clinical tests for measuring biomarkers.

Claims

1. The present invention provides an apparatus for easily and quickly detecting a specific protein in a sample.

2. Protein detection methods and clinical testing methods using this device, as well as antibodies, sample preparation reagents, and detection machines used in this device.

Images