Rapid test paper or paper card for zinc ion colloidal gold chromatography and manufacturing method
A technology for detecting test strips and zinc ions, applied in the fields of immunology and hygiene inspection, can solve the problems of long detection time, difficult adaptation and high cost, and achieve the effects of strong timeliness, easy operation and control, and mild reaction process.
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Embodiment 1
[0049] Example 1. Zn 2+ Synthesis of artificial antigen
[0050] It is called 20mg klh, which is dissolved in N-(2-hydroxye ethyl) -N ’-2-ethylene sulfonic acid buffer (HBS) with a 1ml concentration of 10mmol / L, PH9.0, to form a KLH solution.
[0051] Called 10mg metal octon agent for amino benzhenyl-titthhamine and pentthylene (P-NH2-BN-DTPa), and dissolved in 1ml of two-sectaria (DMSO)The coordinated solution is added to the KLH solution of 1ml under gently stirring, and the pH to 9.0 is adjusted with a 10 mol / L NaOH solution, and the reaction of the room temperature is 24h;Ultrafilled, super filter pipes were washed 3 times with 10 mmol / L, PH9.0 HBS, and then washed 2 times with 10 mmol / L, PH7.4 HBS buffer to remove the unresponsive small molecular metal metal co -agent.
[0052] Take 16 μL concentration to 733.7mmol / L zinc solution drip to the response carrier protein solution, use NaOH solution to adjust its pH value to 9.0, room temperature shake the bed for 24 hours, and ...
Embodiment 2
[0055] Example 2. ZN 2+ Preparation of monoclonal antibodies
[0056] (1) Animal immunity.Immune BALB / C mice, use ZN 2+ -DTPA-KLH is immune to 6 weeks of age female Balb / C mice, with a dose of 20 μg / 0.2ml / piece.Shou -free, dilute Zn with sterilization PBS 2+ -DTPA-KLH, mixed with the equivalent CFA emulsification; strengthening immunity, diluted Zn with sterilization PBS 2+ -DTPA-KLH, mixed with the equivalent IFA, and emulsified for the first 3 weeks after the first free exemption. After that, it is immune once every 2 weeks.30 min, set up overnight at 4 ° C, 4000 R / min 4 ° C centrifugal 5 min, take it to clear, and save for later use.
[0057] (2) Cell fusion spare mouse selection.Indirect ELISA detection 2+ Cheese PAB cost, block ELISA detection Zn 2+ -Dtpa Pab to Zn 2+ -DTPA IC 50 , The highest selection is the highest, IC 50 The lowest mice are used for cell fusion after super exemption.
[0058] (3) Screening of positive hybrid tumor cells.Cell fusion, preheat the PEG-1500...
Embodiment 3
[0061] Example 3. Anti -Zn 2+MAB's identification
[0062] (1) Determination of ascites.Mouse injection of mice after 10 d of paradise for abdominal cavity injection cloning cell strain 10 7 For a cell, take ascites after 10 d, the saturated ammonium sulfate salt analysis method is pure, indirect ELISA measures Zn 2+ MAB's ascites are 1: 1.2 × 10 6 It
[0063] (2) Sensitivity identification.Use block ELISA to measure Zn 2+ MAB to different concentrations zn 2+ -Edta standard liquid inhibitory rate, inhibitory rate B / B 0 For vertical coordinates, use different concentrations ZN 2+ -Edta's pairs of values are horizontal coordinates, draw standard inhibitory curves, perform relevant regression analysis, calculate Zn 2+ mab to Zn 2+ -Edta IC 50 EssenceZn 2+ MAB's linear regression equation is y = -33.827 x + 99.512, R 2 = 0.9891, IC 50 It is 29.08 μg / L.
[0064] (3) Special identification.Adopting cross -reaction tests, selection of mercury, lead, cadmium, copper, chromium, cobalt, ir...
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