Aflatoxin M1 immunochromatographic test strip and preparation method thereof
An immunochromatographic test strip and aflatoxin technology, which is applied in analytical materials, measuring devices, instruments, etc., can solve problems such as the advent of aflatoxin M1 immunochromatographic test strips, and achieve the effect of simple operation and high detection sensitivity.
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Example 1: Preparation method of anti-aflatoxin M1 monoclonal antibody
The anti-aflatoxin M1 monoclonal antibody is produced by the hybridoma cell line 2C9 with the deposit number CCTCCNO.C201018, and its preparation method includes the following steps:
1. Obtain hybridoma cell line 2C9 using a two-step screening method
(1) Animal immunity
Six 6-week-old BALB / c mice were purchased and immunized with the commercially available aflatoxin complete antigen AFM1-BSA. After the first immunization, the complete antigen of aflatoxin and the same amount of Freund's complete adjuvant were emulsified, and then injected into multiple points under the skin of the back of the neck of the mouse. The second immunization was carried out 4 weeks later and was emulsified with Freund's incomplete adjuvant and an equivalent amount of aflatoxin, and injected into the abdominal cavity of the mouse. The third immunization and the second immunization are separated by 4 weeks. The immunization method...
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Example 2-4: Preparation and application of aflatoxin M1 immunochromatographic test strip
The anti-aflatoxin M1 monoclonal antibody used in the following Examples 2-4 is the anti-aflatoxin M1 monoclonal antibody prepared in Example 1.
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Example 2
The method for preparing aflatoxin M1 immunochromatographic test strip as described above includes the following steps:
(1) Preparation of absorbent pad
Cut the absorbent paper to a length of 16mm and a width of 3mm to obtain an absorbent pad;
(2 Preparation of detection pad
Figure 1 is a front view of the aflatoxin M1 immunochromatographic test strip of the present invention. In the picture: 1 cardboard, 2 absorbent pad; 3 inspection pad; 4 gold label pad; 5 sample pad; 6 quality control line; 7 inspection line.
The aflatoxin M1-bovine serum albumin conjugate AFM1-BSA was prepared into a 0.1mg / mL coating solution A; at a position 15mm away from the upper edge of the nitrocellulose membrane, spray the coating solution A Coated horizontally on a nitrocellulose membrane to obtain a detection line. The coating amount of aflatoxin M1-bovine serum albumin conjugate required per centimeter of the detection line is 75ng, and then dried at 37°C for 15 minutes;
The coatin...
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