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Method for preparing prostate cancer antigen immunochromatography test paper based on hydrophobin high-efficiency fixed low-concentration antibody

An immunochromatographic test strip and hydrophobin technology, which is applied in the field of clinical medical detection, can solve the problems of reduced utilization of epitope epitopes, reduced ability to capture antigens, and unsatisfactory detection effects, and can improve the ability to capture antigens and reduce antibodies. The effect of using the content and reducing the cost of testing

Inactive Publication Date: 2017-05-31
TIANJIN UNIV
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  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

During this process, the unsatisfactory detection effect is not due to the denaturation and inactivation of the antibody itself, but the chaotic and unoriented arrangement of the antibodies at the detection line, which leads to the insufficient exposure of the "Fab" end where the antigenic determinant is located, and the use of the epitope of the antigenic determinant of the antibody Therefore, how to improve the detection sensitivity, enhance the antibody immobilization efficiency, and reduce the detection cost has become a major challenge for the current test strip technology.

Method used

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  • Method for preparing prostate cancer antigen immunochromatography test paper based on hydrophobin high-efficiency fixed low-concentration antibody
  • Method for preparing prostate cancer antigen immunochromatography test paper based on hydrophobin high-efficiency fixed low-concentration antibody
  • Method for preparing prostate cancer antigen immunochromatography test paper based on hydrophobin high-efficiency fixed low-concentration antibody

Examples

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preparation example Construction

[0032] Preparation of quantum dot immunoprobe: dissolve water-soluble quantum dots, labeled antibody for prostate cancer, EDC (1-ethyl-(3-dimethylaminopropyl) carbodiimide hydrochloride) in boric acid buffer solution, suspended at room temperature, activated the carboxyl group of the quantum dots, centrifuged, added the same amount of antibody to repeat the above process, and finally the quantum dots coupled with the antibody were blocked overnight with BSA.

[0033] The assembly of the novel test strips with the help of hydrophobin-immobilized antibodies: common test strips are composed of sample pads, binding pads, nitrocellulose membranes, and water-absorbing pads. The test strips of the present invention are closely connected from left to right. It is a sample pad and a binding pad stacked together, in order to fully combine the antigen and the quantum dot-labeled antibody, such as figure 1 As shown, the number of sample pads in this technical invention is two. Different f...

Embodiment example 1

[0046] Preparation method of prostate cancer immunochromatographic test strips based on hydrophobin efficiently immobilizing low-concentration antibodies; 50 μg / ml hydrophobin is sprayed on the detection line position of nitrocellulose membrane, and the mass fraction ratio of quantum dots and EDC consumption is: 1:4000, the mass fraction ratio of quantum dots and antibodies is 1:16, the amount of BSA blocking solution is 1%, and the coating antibody concentration of PSA diluted with 0.01M PBS buffer with pH=7.4 is 0.2mg / ml Spray on the hydrophobin layer, and their overlapping area is used as the detection line area, drop 45 μl of standard antigen with a concentration of 5 ng / ml into the sample pool, do three repetitions, react for 15 minutes, and read the detection with a quantum dot immunofluorescence analyzer The indications of the line and the quality control line are the T value and the C value, and the average value of the T / C intensity of the detection signal obtained is ...

Embodiment example 2

[0048] Preparation method of prostate cancer immunochromatographic test strips based on hydrophobin efficiently immobilizing low-concentration antibodies; 50 μg / ml hydrophobin is sprayed on the detection line position of nitrocellulose membrane, and the mass fraction ratio of quantum dots and EDC consumption is: 1:4000, the mass fraction ratio of quantum dots and antibodies is 1:16, the amount of BSA blocking solution is 1%, and the coating antibody concentration of PSA diluted with 0.01M PBS buffer with pH=8 is 0.2mg / ml Spray on the hydrophobin layer, and their overlapping area is used as the detection line area, drop 45 μl of standard antigen with a concentration of 5 ng / ml into the sample pool, do three repetitions, react for 15 minutes, and read the detection with a quantum dot immunofluorescence analyzer The indications of the line and the control line are the T value and the C value, and the average value of the T / C intensity of the detection signal obtained is 0.165.

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Abstract

The invention relates to a method for preparing prostate cancer antigen immunochromatography test paper based on a hydrophobin high-efficiency fixed low-concentration antibody. As exogenous biomolecules can be subjected to oriented immobilization of type-II hydrophobin, antibodies sprayed to a test paper nitrocellulose membrane can be arranged in an oriented manner, the 'Fab' ends of the antibodies are sufficiently exposed, the utilization rate of an antibody active site can be increased, the detection sensitivity can be improved, the amount of the antibodies can be reduced, the detection cost can be lowered, the nitrocellulose membrane is modified by using the hydrophobin, novel immunochromatography test paper based on the hydrophobin high-efficiency fixed low-concentration antibody can be prepared, the test paper comprises a sample pad and two conjugate pads, a detection line, the nitrocellulose membrane with the detection line and a quality control line antibody, an absorption pad and a bottom plate are simultaneously modified with the hydrophobin, and the method is a novel technique applicable to tumor marker detection.

Description

technical field [0001] The present invention relates to the field of clinical medical detection, and more specifically relates to a method for preparing a novel quantum dot immunochromatography test strip that efficiently fixes the antibody sprayed on the detection line of the test strip by type II hydrophobin and reduces the amount of antibody used in the detection line . Background technique [0002] With economic development, social progress, improvement of people's living standards, changes in behavior and lifestyle, and population aging, people's cause of death spectrum has undergone great changes. Among them, the annual death toll caused by malignant tumors is All are growing, and gradually become the main cause of death of residents, therefore, early diagnosis and treatment of tumors has become a research hotspot in recent years. [0003] Test strip technology is an immunological detection technology that is easy to operate, does not require professional training, is...

Claims

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Application Information

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IPC IPC(8): G01N33/543G01N33/558G01N33/574
CPCG01N33/57434G01N33/54353G01N33/558
Inventor 常津张博宫晓群姚颖异武玉东张健
Owner TIANJIN UNIV
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