Heavy metal copper monoclonal antibody and preparation method thereof
A technology for cloning antibodies and anti-heavy metals, applied in the biological field, can solve the problems of no research reports and a late start, and achieve the effects of good repeatability, high accuracy, and strong antigen practicability
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Embodiment 1
[0044] The detection of embodiment 1 copper standard solution (Sigma Co.)
[0045] The indirect competitive ELISA method was used to conduct a preliminary exploration of the detection curve. The method is as follows:
[0046] Coating: Dilute the coating antigen bovine serum albumin-1-(4-isothiocyanatophenyl)-B with 10mM, pH7.4N-2-hydroxyethylpiperazine-N-2-ethanesulfonic acid buffer Diaminetetraacetic acid-Cu, add 50 μl / well to a 96-well enzyme-linked microanalysis plate, and place it at 4°C overnight;
[0047] Washing: wash 3 times with PBST;
[0048] Blocking: PBST diluted gelatin to 1% (m / v), 100 μl / well, 1.5 hours at 37°C;
[0049] Washing: wash 3 times with PBST;
[0050] Adding samples: dilute the copper nitrate standard solution step by step with phosphate buffer containing 2mM ethylenediaminetetraacetic acid to 10 -5 μg / ml, 10 -4 μg / ml, 10 -3 μg / ml, 10 -2 μg / ml, 10 -1 μg / ml, 1μg / ml, 10μg / ml, 100μg / ml series concentration, the diluted solution was placed at room...
Embodiment 2
[0057] Detect copper residue in the tap water sample of embodiment 2
[0058] Bovine serum albumin-1-(4-isothiocyanatophenyl)-ethylenediaminetetraacetic acid-Cu 0.4 μg / ml coated microanalysis plate 50 μl / well, copper antibody supernatant diluted 4 times as the working concentration, 1% (m / v) gelatin was used as a blocker, the pH value of the antigen-antibody reaction matrix (N-2-hydroxyethylpiperazine-N-2-ethanesulfonic acid buffer solution) was 5.0, and the ionic strength was 0.15mol / L. 50 μl / well in microassay plate.
[0059] Sample preparation to be tested:
[0060] Tap water sample: measure 100 μl of tap water, add 0.5 μl of 1000ug / ml copper standard solution, and prepare a 5ug / ml liquid sample.
[0061] The above samples were detected by copper ELISA detection method, and the operation was as follows:
[0062] 1) Coating:
[0063] Bovine serum albumin-1-(4-isothiocyanatophenyl)-ethylenediaminetetraacetic acid-Cu 0.4μg / ml Add 50μl to each well of the microanalysis plat...
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