Male multi-tumor marker detection protein chip and kit thereof
A tumor marker and protein chip technology, applied in the field of male multi-tumor marker detection protein chip and its kit, can solve the problems of insignificant clinical tests, false positives or false negatives.
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Embodiment 1
[0028] Preparation of a protein chip for detection of multiple tumor markers in males
[0029] The main steps are: (1) Prepare the matrix nitrocellulose membrane; (2) Design the chip, determine the arrangement of the lattice and the spotting position; (3) Draw the antibody from the 384-well plate and spray it on the nitrocellulose (4) assemble a nitrocellulose membrane, and carry out sealing, drying, packaging, and preservation.
[0030] The preparation method of the above-mentioned protein chip for detecting multiple tumor marker antibodies in males specifically includes the following steps:
[0031] 1. Determination of antigen and antibody for tumor marker detection
[0032] Determination of detection index antigen:
[0033] The tumor marker CEA is composed of six peptides with high similarity, so the A1+B1 segment was selected for genetic engineering expression; PSA, NSE, SCC-ag and CYFRA21-1 are full-length genetic engineering expression; while AFP and CA19-9 are purifi...
Embodiment 2
[0041] Kit preparation
[0042] A kit for male multi-tumor marker detection protein chip, composed of male multi-tumor marker detection protein chip, standard product, quality control product, sample diluent, enzyme-labeled working solution, concentrated washing solution, and detection solution; each part For bottled packaging, use a paper template to fix in the box (such as figure 2 ), not limited to the placement method, it is used together with the test. Here's how the parts are prepared:
[0043]1. The male multi-tumor marker detection protein chip is a 6×8-well plate, with nitrocellulose membrane as the base, and the assembly method is shown in ZL032298900. After the completion of the appearance, see figure 1 .
[0044] 2. The standard product provides 6 bottles, each bottle is 0.25ml.
[0045] With 3% BSA and 0.05% preservatives (Proclin TM 300) of phosphate buffer (pH 7.4) configuration, wherein the final concentrations of AFP, CEA, CA19-9, tPSA, NSE, SCC-ag and...
Embodiment 3
[0063] Application and detection of protein chips and kits
[0064] The application of the protein chip for detecting multiple tumor marker antibodies in males according to the present invention mainly adopts the antibody-antigen-enzyme-labeled secondary antibody method to detect tumor marker antibodies in human blood, and the method is to add human serum to the surface of the chip , the antigens in the sample are bound to the corresponding antibodies immobilized on the chip, after drying, add the enzyme-labeled secondary antibody to react, wash away the unbound secondary antibody, scan and collect the signal 1 minute after adding the detection solution, and the sample point The intensity of the signal is proportional to the concentration of the antigen in the serum.
[0065] In the above-mentioned application of the protein chip kit for detection of multiple tumor marker antibodies in males, the specific methods of antigen-antibody reaction and detection are as follows:
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