Immunity probe based on polymer luminous point and preparing method and application thereof
A polymer, luminescent point technology, applied in the field of detection, can solve problems such as heavy metal pollution, and achieve the effect of reliable method, high sensitivity and elimination of interference
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Embodiment 1
[0071] Example 1 Preparation of an immunoprobe based on polymer luminescent dots
[0072] (1) Preparation of imidophos polyclonal antibody
[0073] According to the literature (Xu Ying. Research on immunoassay methods of organophosphorus pesticides dichlorvos and imophos [D]. Huazhong Agricultural University, 2009.) Mannich method was used to couple Phosemet to cationic bovine serum albumin (cBSA) to construct Phosemet complete antigen. According to the literature (Song Yang. Research on the ELISA method of imidophos pesticide residues [D]. Tianjin University of Science and Technology, 2009), the rabbit was immunized with the imothion antigen immunization scheme, and the antiserum titer reached the highest after 66 days. Blood was collected from the heart, rabbit serum was extracted, centrifuged to obtain imidophos antibody, and the concentration range of the antibody obtained by ultraviolet spectrum measurement was 0.64-0.78mg / mL, and stored at -20°C for later use.
[0074]...
Embodiment 2
[0097] Example 2 Stability and Specific Detection of Immunoprobes Based on Polymer Luminescent Dots
[0098] (1) According to the optimal reaction conditions of Example 1, the immune probe based on the polymer luminescent point was prepared. The optimal reaction conditions were as follows: the reaction mass ratio of PDs to antibodies was 4:5, the reaction temperature was 37°C, and the reaction time was 2h. The pH value of the system is 7.3. The prepared immune probe (PDs-Ab solution) was tested in the following steps.
[0099] (2) Detection of stability: store the immunoprobe based on the polymer luminescent point at 4°C. During the first week of storage, there was little change in fluorescence intensity. After that, it was measured once every other week, and the fluorescence intensity decreased slightly with the increase of time, and after 30 days of storage, the fluorescence intensity decreased by 10%. The above results show that the fluorescent properties of the probe ar...
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