Antibody chip reagent kit for detecting hepatoma marker
A technology of antibody chips and markers, applied in the biological field, can solve the problems of low tumor sensitivity and specificity, and achieve the effects of less specimen consumption, saving sample volume, consistency and reliability time
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Embodiment 1
[0040] Example 1: Preparation of the kit.
[0041] 1. Antibody screening and source
[0042] Taking AFP antibody screening as an example, this antibody is used to detect 6 clinical samples (the detection value of the clinical sample is known and measured by electrochemiluminescence method), and the detection value of the clinical sample is compared with the detection value of this kit to draw a scatter diagram , and its correlation is as follows: the detection result of electrochemiluminescence is basically equivalent to the detection result of this kit.
[0043] Taking the AFP antibody as an example, this antibody was used to detect 6 clinical samples (the detection value of the clinical sample is known, measured by electrochemiluminescence method, which is the light absorption peak value converted into a value by software), and the detection value of the clinical sample was compared with this reagent. The detection value of the box is compared and drawn as a scatter plot (s...
Embodiment 2
[0079] Example 2: The kit described in the present invention is used in the experiment of detecting liver cancer tumor markers.
[0080] 1. Complete drying of the slide chip
[0081] Take out the antibody chip from the liver cancer marker detection antibody chip kit of the present invention.
[0082] There are two types of frame for the antibody chip. If one antibody is spotted with 4 wells (ie 4 repetitions), a 2×8 chip frame can be used; if an antibody is spotted with 2 wells, a 4×16 chip frame can be used.
[0083] After equilibrating the slide chip at room temperature for 20-30 minutes, open the packaging bag, peel off the sealing strip, and then place the chip in a vacuum desiccator or dry at room temperature for 1-2 hours.
[0084] 2. Chip operation process
[0085] Add 50-100 μl of sample to each well, and the sample volume of different samples is different: plasma and serum are diluted 1:1 with sample diluent before use; cell supernatant can be stock solution; cell o...
Embodiment 3
[0104] Example 3: The detection sensitivity of the antibody used in the kit of the present invention and the cross-reaction result of the antibody
[0105] The detection sensitivity of the antibody used in the test kit of the present invention
[0106] The specific operation refers to Example 2, and the minimum detection limit and linear range of the kit are obtained by serially diluting each antigen.
[0107] Table 2: The detection sensitivity of 8 kinds of antibodies in the kit of the present invention
[0108]
[0109] Two, the cross reaction result of the antibody used in the kit of the present invention
[0110] The specific operation refers to Example 2, and the detection antibodies are anti-AFP antibody, anti-CK19 antibody, anti-VEGF antibody, anti-DCP antibody, anti-SCCA antibody and the mixture of these five antibodies. The cross-reactivity test results of the antibody pairs are as follows: Figure 9 shown.
[0111] It can be seen from the results that each ant...
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