Lung cancer marker detection immunochromatographic test paper and application
An immunochromatographic test strip and marker technology, applied in measurement devices, analytical materials, instruments, etc., can solve the problem that the sensitivity and specificity of tumor markers cannot meet the requirements of early diagnosis, differential diagnosis, curative effect and prognosis evaluation, etc. High sensitivity, simple test operation and accurate test results
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Embodiment 1
[0058] Preparation of colloidal gold-neuron-specific enolase ((NSE)) antibody complex:
[0059] Utilize the colloidal gold of newly prepared 40nm to connect with (NSE) antibody, concrete implementation steps are as follows:
[0060] 1. Take 1mL colloidal gold solution, add 10uL 0.1M K 2 CO 3 Adjust the pH of the solution to 9.0 and mix well.
[0061] 2. Add 8uL (NSE) monoclonal antibody (L1C00501) into the colloidal gold solution, mix well under vortex, and let stand for 10min.
[0062] 3. Add 200uL 10% BSA solution, mix well, and let stand for 10min.
[0063] 4. Centrifuge the colloidal gold solution obtained above at 10,000 rpm for 15 min under refrigeration.
[0064] 5. Aspirate the supernatant, redisperse the precipitate with 1 mL borate buffer solution (containing 0.5-1.0% sucrose, 2-5% BSA, 0.01M boric acid), and then centrifuge once at high speed.
[0065] 6. Aspirate the supernatant, concentrate to 1 / 10 of the original volume, and store at 4°C.
Embodiment 2
[0067] Preparation of colloidal gold-carcinoembryonic antigen ((CEA)) antibody complex:
[0068] Utilize the colloidal gold of newly prepared 40nm to connect with (CEA) antibody, concrete implementation steps are as follows:
[0069] 1. Take 1mL colloidal gold solution, add 10uL 0.1M K 2 CO 3 Adjust the pH of the solution to 9.0 and mix well;
[0070] 2. Add 10uL (CEA) monoclonal antibody (LLC00202) into the colloidal gold solution, mix well under vortex, and let stand for 10min;
[0071] 3. Add 200uL 10% BSA (bovine serum albumin) solution, mix well, and let stand for 10min;
[0072] 4. Centrifuge the colloidal gold solution obtained above at 10,000 rpm for 15 min under refrigeration;
[0073] 5. Aspirate the supernatant, redisperse the precipitate with 1mL borate buffer solution (containing 0.5-1.0% sucrose, 2-5% BSA, 0.01M boric acid), and then centrifuge once at high speed;
[0074] 6. Aspirate the supernatant, concentrate to 1 / 10 of the original volume, and store at ...
Embodiment 3
[0076] Preparation of double antibody sandwich (NSE), (CEA) test strips:
[0077] 1. Preparation of the sample pad: select glass cellulose membrane as the sample pad material, cut into strips of 5.0×30.0 cm in size, put it into the sample pad blocking solution 0.01mol / L PBS (phosphate buffer solution) (PH=7.4 ), 1.0-2.0% BSA (bovine serum albumin); 2.0-2.0% sucrose, 0.1-0.5% Tween, 0.1-1.0% PVP (polyvinylpyrrolidone)) for 30 minutes, and dried at 37°C for later use.
[0078] 2. Preparation of conjugation pads: glass cellulose membrane was selected as the conjugation pads, and cut into strips with a size of 1.0×30.0 cm for later use.
[0079] 3. Preparation of NC (nitrocellulose membrane): stick the NC membrane on the bottom plate, and draw (CEA), (NSE) monoclonal antibody and goat anti-mouse IgG antibody on different positions on the NC membrane with a scriber, as a detection tape and quality control tape.
[0080] 4. Assembly of immunochromatography test strips: Paste absor...
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