Antibody-based drug for treating psoriasis
A technology for antibody drugs and psoriasis, which is applied in the direction of antibodies, drug combinations, skin diseases, etc., can solve the problems of no research on antibodies, and achieve the effect of simple preparation method and no toxic side effects
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Embodiment 1
[0029] Pathogen Isolation
[0030] 1. After conventional 75% alcohol disinfection of the psoriasis patients, the epidermal cells (dander) and deep epidermal cells were scraped from the affected area (patient specimens were provided by Shian Clinic and Haixin Clinic). 11 kinds of samples were obtained from the affected area. The 11 samples were respectively inoculated on the slant of Sabouraud dextrose agar containing 0.05% chloramphenicol, and cultured at 28°C for 2 weeks. Sabouraud dextrose agar was prepared according to Chinese Pharmacopoeia.
[0031] 2. Pick the cultured fungi, inoculate them on the Sabouraud dextrose agar plate by the streak separation method, and culture them at a temperature of 28°C until a single colony is cultivated. Carry out visual observation and record the state of each colony, determine the yeast-like bacteria and mold, and then stain the mold with cotton blue to make a smear to observe the spore morphology under the light microscope, combine th...
Embodiment 2
[0033] Prepare antigen
[0034] a bacteria culture, pick 4 kinds of fungi and inoculate them into test tubes containing 5ml potato dextrose medium, place them in a 28°C, 160rpm shaking incubator for 36h (24-48h can be achieved), and then transfer to 100ml potato dextrose culture In a base Erlenmeyer flask, culture in a 28°C, 160rpm shaking incubator for 60h (48-72h can be achieved). The liquid culture was collected, centrifuged at 6000rpm for 10min, and the supernatant was discarded.
[0035] b Formaldehyde inactivation, add 3.7% formaldehyde to the precipitate after centrifugation, overnight at 4°C, centrifuge under the same conditions, wash with normal saline 3-4 times.
[0036] c Inactivation verification: pick inactivated bacteria respectively, streak and smear on Sabao weak glucose agar medium, culture at 28°C for 60h (48-72h can be achieved), aseptic growth and inactivation are successful.
[0037] d Antigen preparation, the sterilized bacteria were resuspended in phys...
Embodiment 3
[0039] Antibody preparation
[0040] To immunize rabbits, for the first immunization, take 1ml of protein solution (containing 0.5mg of total protein) and mix it with an equal volume of Freund's complete adjuvant, and grind it in a mortar until the emulsion does not disperse when it enters water, indicating that the emulsification is complete. The preparation method of the antigen for booster immunization is the same as above, as long as Freund's incomplete adjuvant is used instead of Freund's complete adjuvant. Antigens were added on days 15, 30, and 45 for further immune boosting, and antiserum was collected within one week after the last injection of immune boosting.
[0041] 5. Determine the titer of the antibody, and use the indirect ELISA method to measure the titer of the polyclonal antiserum. Before immunization, 1ml of blood was taken from the rabbit's ear vein as a negative control. Before the fourth immunization, 1ml of blood was taken from each rabbit to measure t...
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