Colloidal gold chromatography anti-Jo-1 antibody detection test paper and preparation method thereof
A technology of antibody detection and colloidal gold, which is applied in the field of medical immunization and can solve the problem that anti-Jo-1 antibody has not come out
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Embodiment 1J
[0071] Embodiment 1Jo-1 antigenic protein preparation
[0072] The Jo-1 antigen protein used in this test paper was constructed by gene cloning technology to construct a recombinant gene, and then the Jo-1 antigen protein which was all human was successfully expressed by prokaryotic expression technology. Wherein, the Cenp-B antigenic protein is derived from purified genetically engineered antigenic protein.
Embodiment 2
[0073] Example 2 Antibody Preparation
[0074] Antibody A, Antibody C, and Antibody B were prepared by the following method. The anti-human IgG in antibody A, antibody C and antibody B can generally be produced by multiple subcutaneous (sc) or intraperitoneal (ip) injections of purified immunogens and adjuvants on animals.
[0075] The injection solution is obtained by mixing 0.05mg~1mg of immune preparations (for goats or mice respectively) with 3 times the volume of Freund's complete adjuvant, and the injection solution is injected into multiple parts of the animal skin. After one month, the animal is treated with 1 / 5 to 1 / 10 of the original amount of human IgG in Freund's complete adjuvant mixture was injected subcutaneously into multiple sites to boost the immunization. After 7-14 days, the animals were bled, and the serum anti-human IgG titer was measured. Animals were boosted until titers plateaued. Methods for producing polyclonal antibodies are described in many im...
Embodiment 3
[0078] Colloidal gold solution preparation
[0079] 0.01% HAuCl 4 Heat the solution to boiling, quickly add every 100mL HAuCl 4 Add an appropriate amount of reducing agent solution to the solution, and the color changes from blue, then light blue, to blue, then red after heating, and transparent orange-red after boiling for 7-10 minutes. Then filter with ultrafiltration or microporous membrane (0.45μm) to remove polymers and other possible impurities. The prepared colloidal gold should be pure, translucent, free of sediment and floating matter, and discard when oily matter and a large amount of black granular precipitate appear on the liquid surface.
[0080] Wherein the reducing agent used can be trisodium citrate (Frens1973), tannic acid-trisodium citrate (Slot and Gueeze1985), white phosphorus, preferably use trisodium citrate, more preferably use 1% trisodium citrate.
[0081] The glass containers used should be absolutely clean, and must be pickled and siliconized befo...
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