Method for preparing optical sensitive film for fixing human brain natriuretic peptide (BNP)
A technology of brain natriuretic peptide and sensitive membrane, applied in the field of clinical medical diagnosis, can solve the problems of increased detection cost and achieve the effect of rapid detection
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[0029] The preparation method of the optically sensitive film for fixing human brain natriuretic peptide of the present invention includes the following steps:
[0030] 1. Preparation of a conjugate of fluorescent quantum dots and brain natriuretic peptide detection antibody, which can recognize the inner ring structure of brain natriuretic peptide molecule.
[0031] 2. Select the carrier. The carrier can be a biological slide or ordinary glass or PC board or organic polymer film.
[0032] Such as figure 2 Shown:
[0033] 3. Surface treatment: Use silane as a surfactant to aminate the surface of the glass sheet. The amino group can be combined with the bifunctional reagent glutaraldehyde, and the glutaraldehyde can be coupled with the capture antibody to achieve the immobilization of the brain natriuretic peptide capture antibody.
[0034] 4. Immobilization of the antibody: Add the brain natriuretic peptide capture antibody that can recognize the C-terminal region of the brain natriur...
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[0039] Examples:
[0040] 1) Preparation of conjugate of fluorescent quantum dots and brain natriuretic peptide antibody
[0041] Use a pipette to transfer 20μl (2nmol / L) avidin-labeled cadmium selenide (CdSe) fluorescent quantum dots into a 2ml centrifuge tube, and add 100μl Tris-HCl buffer (containing 1mmol / LEDTA, 1mol / L) NaCl, pH=7.4), 10 μl of biotin-labeled brain natriuretic peptide antibody was dissolved in 80 μl of Tris-HCl buffer, and the antibody concentration was 0.15 mg / mL at this time. The above-mentioned biotin-labeled antibody solution was added to the avidin-modified fluorescent quantum dot solution, and placed in a shaker at 37° C. and 150 rpm / min to react for 30 minutes. 1ml of PBST buffer (containing 0.05% Tween-20 and 0.1% BSA) was added, and the brain natriuretic peptide antibody solution that did not participate in the reaction was removed by centrifugation. PBS buffer was added to prepare a fluorescent quantum dot-labeled brain natriuretic peptide antibody s...
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