Colloidal gold test strip for double-amplification system and preparation method thereof
A technology of colloidal gold test paper and amplification system, which is applied in biological testing, material inspection products, measuring devices, etc., can solve the problem of not further improving the sensitivity of colloidal gold, and achieve the effect of enhancing binding force, improving specificity, and improving sensitivity
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[0022] Example 1
[0023] Such as figure 1 As shown, a colloidal gold test strip of a bi-amplification system includes a test strip bottom liner 1, and a sample pad 2 on the test strip bottom liner 1 that are sequentially overlapped and pasted with each other. Ester film 3, coating film 4 and absorbent paper 5, the coating film 4 is coated with a detection line 6 and a control line 7, and the middle of the coating film 4 is paralleled with a streptavidin-cardiac troponin I monoclonal antibody F(ab) of 4C2 2 Fragment detection line 6 and a control line 7 coated with rabbit anti-mouse IgG antibody, sample pad 2 is coated with F(ab) of mouse-derived cardiac troponin I monoclonal antibody 4C2 2 Fragment-Biotin, gold-labeled antibody is a gold particle labeled ProteinA, which non-specifically binds to the Fc segment of cardiac troponin I monoclonal antibody M155, and indirectly binds to the gold by the amplification of cardiac troponin I monoclonal antibody M155 On the particles. Th...
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[0045] Example 2 The specific preparation method of the colloidal gold test strip using the conventional method is as follows:
[0046] Cardiac troponin I monoclonal antibodies 4C2 and M155 (pairing antibodies) were purchased from Fitzgerald.
[0047] Preparation of coating film:
[0048] 1) Preparation of coating buffer: filter 0.025M, pH7.4 PBS with a 0.22u membrane, and place it at 4°C for use. The validity period is 7 days.
[0049] 2) Preparation of blocking solution: filter PBS containing 1% BSA, 1% sucrose, 0.025M, and pH 7.5 with a 0.22U membrane, and place it at 4°C for use. The validity period is 3 days.
[0050] 3) F(ab) of monoclonal antibody 4C2 2 Preparation of the segment: the monoclonal antibody was digested with trypsin, passed through the SPA-Sepharose CL-4B gel chromatography column to remove the Fc fragment, and collected F(ab) 2 Fragment.
[0051] 4)F(ab) 2 The preparation of section detection line 6: The antibody is at a concentration of 4mg / ml, the peristaltic pump...
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