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Preparation method of antibody-based drug used 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

Inactive Publication Date: 2015-12-02
TUKE TIANJIN PHARMA TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, taking specimens from the affected part of the patient and culturing them to isolate the fungus to treat the disease as an antigen and then to obtain antibodies has not been studied.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

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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Abstract

The invention provides a preparation method of an antibody-based drug used for treating psoriasis. A pathogenic bacterium is directly used as an antigen for immunization, and remarkable antibody effect is acquired; the pathogenic bacterium is an existing bacterial strain, a dander sample can also be obtained from an affected part of a patient with psoriasis, the sample is cultured on a Sabouraud's dextrose agar medium containing chloramphenicol to obtain fungi, culturing by streaking is performed to obtain a single colony, a culture is verified, the single colony is selected and inoculated to a liquid medium and subjected to amplified cultivation, and centrifuging is performed to collect bacteria, formaldehyde inactivating is performed to prepare an antigen, the antigen is used to directly immunize a new Zealand white rabbit to obtain an antibody, and the drug prepared mainly with the antibody has remarkable alleviating effect for treating the patient with psoriasis; the preparation method is simple and has no toxic or side effect, a targeted medicament can be prepared according to pathogenic bacteria of a single patient and used for purposeful targeted treatment.

Description

technical field [0001] The invention relates to the field of drugs for treating psoriasis, in particular to a method for preparing antibody drugs for treating psoriasis. Background technique [0002] Psoriasis is a common chronic inflammatory skin disease that people suffer from, and its etiology is still unclear. There are four types of psoriasis, of which psoriasis vulgaris is the most common. Typical lesions are well-demarcated erythema and papules covered with dry, multilayered scales. The course of the disease is long, the state of an illness recurs, and the clinical treatment is relatively difficult. At present, there are many drugs for the treatment of psoriasis, but because of its complicated etiology, the curative effect of many drugs can not be determined, and the main purpose is to improve the symptoms of patients. Traditional therapeutic drugs have many adverse reactions. In contrast, anti-psoriasis biological agents have the advantages of strong pertinence, d...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/42A61P17/06C07K16/14C07K16/06
Inventor 张耀洲赵娜徐嵩淼张玉环
Owner TUKE TIANJIN PHARMA TECH CO LTD
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