Preparation method for quantum dot-cardiac troponin I antibody immune complex, and preparation method for test strip
A technique for cardiac troponin and immune complexes, which is applied in the field of biomarkers, can solve the problems of unable to obtain a standard curve of myocardial markers, unable to realize the detection of myocardial markers, etc., and achieves good coupling effect, simple preparation method, and standard curve. Good results
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Embodiment 1
[0045] Preparation of a quantum dot-cardiac troponin I antibody immune complex:
[0046] Dissolve 5ul of quantum dots (average particle size of 18 nm, emission peak of 625nm±5nm, concentration of 8uM, purchased from Wuhan Jiayuan) in 500ul, pH 5.5, 0.01M borate buffer, vortex and mix for 30s . Now prepare 0.01M 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride and 0.01M N-hydroxysulfosuccinimide, the solvent is pH 5.5, 0.01M boric acid salt buffer. Add 0.8 ul, 0.01M 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride to the quantum dot solution, vortex and mix, add 40ul, 0.01M N- Hydroxysulfosuccinimide, vortexed to mix. Sonicate in ice bath for 30min. After the activation is completed, take out the activation reaction solution, place it in an ultrafiltration tube (100kd), and centrifuge in a low-temperature centrifuge at a speed of 3500 rpm for 7 minutes. After completion, add 1ml of borate buffer, pH 8.5, 0.01M, to the reaction solution solution, rep...
Embodiment 2
[0056] Basically the same as in Example 1, the difference is that: when preparing the quantum dot-cardiac troponin I antibody immune complex, borate buffers with different pH were used to adjust the pH of the reaction solution after activation, and the agarose gel Electropherogram see figure 2 , wherein, 1 is the band of the quantum dot-cardiac troponin I antibody immune complex prepared by the borate buffer solution of pH 5.5, and 2 is the quantum dot- The band of the immune complex of cardiac troponin I antibody, 3 is the band of the quantum dot-cardiac troponin I antibody immune complex prepared in borate buffer solution at pH 7.5, and 4 is the boron at pH 8.5 The bands of the quantum dot-cardiac troponin I antibody immune complex prepared in salt buffer solution, 5 are the bands of the quantum dot-cardiac troponin I antibody immune complex prepared in the borate buffer solution of pH9.5 Band, 6 is the band of the control.
[0057] Analysis Due to the different isoelectr...
Embodiment 3
[0060] Basically the same as Example 1, the difference is: when preparing the quantum dot-cardiac troponin I antibody immune complex, respectively adding different molar ratios of quantum dots (QDs) and 1-ethyl-3-(3- Dimethylaminopropyl) carbodiimide hydrochloride (EDC), agarose gel electrophoresis image 3 , wherein, 1 is the band of contrast; 2 is the band that the molar ratio of quantum dots and 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride is 1:200, 3 is the strip with the molar ratio of quantum dots and 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride being 1:1000, and 4 is the band of quantum dots and 1-ethyl- 3-(3-Dimethylaminopropyl)carbodiimide hydrochloride is a band with a molar ratio of 1:5000, 5 is quantum dots and 1-ethyl-3-(3-dimethylaminopropyl) The molar ratio of carbodiimide hydrochloride is 1:25000, and 6 is the band of quantum dots and 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride. Bands at a molar ratio of 1:5000...
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