IL-6 immunoturbidimetry detection kit prepared based on recombinant monoclonal antibody, and preparation method thereof
A technology for detection kits and monoclonal antibodies, applied in the fields of genetic engineering and immunological determination and analysis, can solve the problems of high cost, poor detection specificity, difficulty in popularization and application of chemiluminescence detection and time-resolved fluorescence chromatography, and achieve improved Detection sensitivity, good specificity, and the effect of improving application value
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Embodiment 1
[0043] Preparation of anti-IL-6 latex microsphere mother solution:
[0044] 1. Cleaning: Take latex microspheres with a particle size of 300nm into a 50ml centrifuge tube, add 10 times the volume of 20mMMES activation buffer, shake and mix, and centrifuge at 20000r / min for 30min.
[0045] 2. Activation: Add the same volume of activation buffer, blow and mix the microspheres with a Tip suction head, transfer to a 15ml centrifuge tube, and ultrasonically mix. Ultrasonic parameters are: power 60W, work for 1s, rest for 1s, total time 2min, 3 cycles of ultrasound for backup.
[0046] 3. Combine: Weigh 1.5 times the required amount of EDC, put it into a clean centrifuge tube, and dissolve it in 1ml of pure water. Add EDC to the sonicated microsphere suspension as needed, and shake for 30s. Place in a 37°C water bath and shake for 25 minutes. After activation, transfer the liquid to a 50ml centrifuge tube and centrifuge at 20000r / min for 30min. Add the same volume of binding buffe...
Embodiment 2
[0051] Preparation of IL-6 Immunoturbidimetric Kit Calibrator:
[0052] The IL-6 calibrator contains:
[0053]
Embodiment 3
[0054] Embodiment 3: Preparation of IL-6 immunoturbidimetric assay kit:
[0055] The IL-6 detection kit includes reagent R1 and reagent R2:
[0056] (1) Reagent R1 contains:
[0057]
[0058] (2) Reagent R2 contains:
[0059]
[0060] (3) Assay method of the kit:
[0061] Analysis method: two-point endpoint method;
[0062] Reaction direction: ascending reaction;
[0063] Calibration method: Logit-Log (4P) or SPLINE processing;
[0064] Measurement wavelength: 600nm;
[0065] Measuring temperature: 37°C;
[0066] Sample: Reagent R1: Reagent R2 = 8:200:40 (μL)
[0067] Test steps: Take 8 μL of sample, add 240 μL of reagent R1, incubate at 37°C for 3-5 minutes, add 60 μL of reagent R2, read the absorbance value A1 after 1 minute, read the absorbance value A2 after 5 minutes, and calculate ΔA.
[0068] Calibration method: 6-point calibration, use Hitachi 7180 automatic biochemical analyzer (or other brand models) for detection, set the concentrations of the calibrat...
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