Fourth-generation HIV (Human Immunodeficiency Virus) antibody antigen test paper
A technology for antigen detection and antigen, which is applied in the field of fourth-generation HIV antibody antigen detection test strips and its preparation, can solve the problems of restricting the use of ordinary users, greatly affecting the detection effect of operator proficiency, and being inconvenient for users to use. Promotion, easy to use, simple operation effect
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Embodiment 1
[0032] Embodiment 1: the fourth generation HIV antibody antigen detection test paper (see figure 1 and figure 2 )
[0033] (1) Preparation of P24 Antigen Monoclonal Antibody
[0034] The ascites of P24 antigen monoclonal antibody was obtained by the general method of producing monoclonal antibody in animals, and then the ascites of P24 antigen monoclonal antibody was purified. After ammonium precipitation, use ultraviolet spectrometry to measure protein concentration, use SDS-PAGE to measure purity, the protein concentration of the obtained P24 antigen monoclonal antibody is higher than 2.0mg / ml, and the purity is higher than 90%.
[0035] (2) Preparation of GP36, GP41 and GP120 antigens
[0036] Precipitate the bacterial culture solution containing GP36, GP41 or GP120 antigen with caprylic acid-saturated ammonium sulfate, then purify it by ion chromatography column chromatography, and measure the protein concentration by UV spectrophotometry, and measure the purity by SDS...
Embodiment 2
[0054] Embodiment 2: detection method (see image 3 , 4 、5)
[0055] Add 60 to 100 μl of the sample to be tested (whole blood, serum or plasma) directly to the sample pad 4 in the test paper obtained in Example 1 (the whole blood sample needs to add some additional diluent), and the sample will be along the reaction support 5 Each attachment moves towards the absorbent filter paper, and the experimental results are read within 20 minutes.
[0056] result:
[0057] refer to image 3 , if the sample contains the P24 antigen, it will specifically bind to the colloidal gold-labeled monoclonal antibody to the P24 antigen on the detection test paper to form a complex, and continue upward along the membrane area to form a complex with another coated nitrocellulose membrane 2 A monoclonal antibody to the P24 antigen specifically binds, and red or pink appears at the band T1; if the sample contains HIV antibodies, it is specific to the colloidal gold-labeled monoclonal antibodies ...
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