Fast detection test paper tape of lead ion aurosol immune layer as well as preparation method and application thereof
A detection test paper, immunochromatography technology, applied in the field of immunology, to achieve the effect of low cost, simple operation, and guaranteed benefits
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Embodiment 1
[0046] (1) Preparation and purification of anti-lead ion monoclonal antibody: use bifunctional diethylenetriaminepentaacetic acid (p-SCN-Bn-DTPA) chelating agent to couple lead ions to bovine serum albumin (BSA), mix BALB / c mice were immunized with incomplete adjuvant, the splenocytes of the immunized mice were fused with SP2 / 0 myeloma cells, and positive cell lines stably secreting anti-lead ion monoclonal antibody (MAb-Pb) were screened out and expanded for culture. Inject cells into mice to induce ascites; take 1 volume of ascites, add 2 volumes of 0.06mol / L sodium acetate buffer solution with pH5.0 (59ml sodium acetate solution with a concentration of 0.06mol / L + 41ml with a concentration of 0.06mol / L L of glacial acetic acid mixed), then adjust the pH to 4.5 with 1mol / L hydrochloric acid; slowly add octanoic acid with a volume ratio of 33ul / ml dropwise within 30min under stirring at room temperature, let stand at 4°C for 2h, then 4°C Centrifuge at 10000rpm / min for 30min t...
Embodiment 2
[0055] To verify the detection specificity test of the present invention:
[0056] The lead ion colloidal gold immunochromatographic rapid detection test strip that following heavy metal utilization embodiment makes carries out specificity cross reaction test, and the result is as follows:
[0057] Table 1 Cross-reactivity of other heavy metal lead
[0058]
[0059] The results showed that the antibody except for Pd 2+ Besides, there is almost no cross-reaction to several other metals, so this colloidal gold test strip can be used to detect Pd 2+ .
Embodiment 3
[0061] Verify reliability test of the present invention: (see step b and c image 3 shown, step d see Figure 4 shown)
[0062] a. Prepare Pd first 2+ A series of gradient standard solutions (10, 20, 50, 80, 100 ng / mL) each 50 μL, and then add 50 μL 1mmol / L DTPA solution as a chelating agent, mix well, so that lead ions and DTPA are fully chelated into Pd-DTPA, as The sample solution to be tested.
[0063] b. Use a dropper to draw 80-100 μL of the sample solution to be tested obtained in step a, drop it on the sample pad of the test strip, and start timing after dropping the sample;
[0064] c. Read the result 3 to 5 minutes after adding the sample. When reading, place the test strip vertically in front of the observer with one end of the sample pad facing down;
[0065] d. Judgment of results: C line is displayed as a red line. When the T line develops color, the result is negative, and the lead ion concentration in the sample to be tested is less than 50ng / ml; when the T...
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