Multi-antibody immunomic mass spectrum kit for liver cancer marker
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A kit and marker technology, applied in the multi-antibody immunoassay mass spectrometry kit and its detection field, can solve the problems of high price and unfavorable promotion, and achieve the effects of low cost, simplified preparation process, and enhanced enrichment specificity.
Active Publication Date: 2012-06-20
BEIJING C & N INT SCI TECH +1
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However, this system has the same disadvantages as expensive and unfavorable for promotion as SELDI-TOF-MS
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Embodiment 1
[0032] Example 1 Preparation method of anti-synthetic peptide 6 polyclonal antibody
[0033] 1) Using synthetic polypeptide 6 coupled with carrier protein KLH as an immunogen to immunize white rabbits; its full-length sequence is: NLGHGHKHDRDHGHGHQ (SEQ ID No.1).
[0034] 1) Using synthetic polypeptide 6 coupled with carrier protein KLH as an immunogen, combined with Freund's complete adjuvant and incomplete adjuvant to immunize big-eared white rabbits;
[0035] 2) After 3-4 weeks, take blood from the ear vein to test the titer, and the titer is above 1:50000 to meet the standard of blood collection;
[0036] 3) Take blood from the carotid artery to obtain polyclonal antibody serum;
[0037] 4) The polyclonal antibody is purified by affinity chromatography.
Embodiment 2
[0038]Example 2 Preparation method of polyantibody immunoassay kit
[0039] Take 20 μl of protein A-coated agarose particles (Protein A Agarose, Santa Cruz), put it in a 0.2 mL Eppendorf tube, add 7.5 μg of the polyclonal antibody prepared in Example 1, rotate at 4°C (speed 5r / min), and mix for 1 After one hour, let stand for 2 minutes, remove the supernatant, and wash the obtained solid-phase carrier precipitate with 100 μL of PBS buffer solution (0.01 mol / l, pH 7.4) for 3 times.
Embodiment 3
[0040] Example 3 Preparation method of polyantibody immunoassay kit
[0041] Take 25 μl of protein A-coated agarose particles (Protein A Agarose, Santa Cruz), put it in a 0.2mL Eppendorf tube, add 24 μg of the polyclonal antibody prepared in Example 1, rotate at 4°C (speed 5r / min), and mix for 2 hours , stand for 3 minutes, remove the supernatant, and wash the obtained solid-phase carrier precipitate 5 times with 100 μL of PBS buffer solution (0.01 mol / l, pH 7.4).
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Abstract
The invention relates to a multi-antibody immunomic mass spectrum kit for a liver cancer marker. The kit comprises serum polypeptide polyclonal antibodies and a buffer solution; the polyclonal antibodies are fixed on solid-phase carriers which are protein A or protein G agarose particles; the serum polypeptide polyclonal antibodies are anti synthetic peptide 6 polyclonal antibodies; and the full-length sequence of synthetic peptide 6 is shown as SEQ ID No.1, namely NLGHG HKHDR DHGHG HQ. Multiple kinds of polypeptide markers of liver cancer are selected; multiple kinds of polyclonal antibodies are prepared correspondingly; through detection and analysis and data statistics, the detection specificity and sensitivity of one kind of polyclonal antibodies are higher than those of other kinds of the polyclonal antibodies, so the marker has high diagnostic capability of liver diseases; and therefore, the immunomic mass spectrum kit containing multiple kinds of the polyclonal antibodies, and a detection method are provided.
Description
technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a polyantibody immune mass spectrometry kit and a detection method thereof. Background technique [0002] Primary liver cancer is one of the cancers with the highest incidence in the world. The five-year survival rate of malignant tumors diagnosed early can reach 70-95%, while the five-year survival rate of advanced tumors is only 20-30%. Under the premise that the treatment methods cannot be improved quickly, improving the early diagnosis rate of malignant tumors to achieve early detection and early treatment of malignant tumors is an effective means to improve the overall survival rate of malignant tumor patients, improve the quality of life, and reduce medical expenses. Currently, commonly used clinical diagnostic methods include physical examination, imaging examination, serum tumor marker detection and pathological examination. Due to the small size of the cancer in...
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