Liquid aerosol lung delivery immunity mouse model of botulinum neurotoxin AHc vaccine
A botulinum neurotoxin and aerosol technology, which is applied in the fields of antitoxins, muscular system diseases, and antibody medical components, etc., can solve the problems of large size of instruments and equipment, inability to accurately quantify the dose, and large loss.
Pending Publication Date: 2019-08-20
ACADEMY OF MILITARY MEDICAL SCI
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Problems solved by technology
At present, the research on liquid aerosol or dry powder aerosol mostly uses mouse mouth and nose exposure and whole body exposure models. Although these two methods are non-invasive to experimental animals, they cannot accurately quantify the dosage, and the samples are lost in the exposure tower. Large and prone to subsidence
Moreover, the related instruments and equipment are bulky and expensive.
Method used
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Embodiment 1
[0054] Embodiment 1, the preparation of albumen
[0055] 1. Preparation of botulinum neurotoxin type A (BoNT / A)
[0056] Preparation of botulinum neurotoxin type A. It was prepared according to the method recorded in the literature ("Establishment of a Convenient and Easy-to-Use Method for Detecting the Activity of Botulinum Toxin Type A", Liu Jing). Using Clostridium botulinum typeA 62A, first enrichment and toxin production culture, then precipitation and extraction, then ammonium sulfate precipitation (60% saturation) and nucleic acid removal, and then gel Filtration and ion exchange chromatography to obtain botulinum neurotoxin type A solution. See the literature for specific steps.
[0057] Take botulinum neurotoxin type A solution, and measure the protein concentration by BCA method, the protein concentration is 0.15mg / mL.
[0058] 2. Preparation of AHc protein
[0059] The AHc protein is the nontoxic 50 kD carboxy-terminal fragment of the heavy chain of botulinum n...
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Abstract
The invention discloses a liquid aerosol lung delivery immunity mouse model of a botulinum neurotoxin AHc vaccine. The effective component of an A type botulinum neurotoxin vaccine provided by the invention is aerosol of a specific solution, wherein the specific solution contains specific protein and a mucosal immunity adjuvant, the specific protein is AHc protein or fusion protein having the AHcprotein, the mass compounding ratio of the specific protein to the mucosal immunity adjuvant is 1 to (0.5-1.5), and the AHc protein is the protein as shown in a sequence 1 of a sequence table. A recombinant protein AHc subunit vaccine is selected, a lung delivery immunity manner is used for the first time, and in vivo efficacy and in vitro efficacy of the AHc subunit vaccine are researched. The vaccine can protect a mouse from being attacked by a deadly A type botulinum neurotoxin. Lung delivery is performed through trachea, so that samples can directly reach the lung target organ, the effectsare direct, medicines are saved, and a new technical means is provided for inhalation infection and treatment.
Description
technical field [0001] The invention relates to a liquid aerosol lung delivery immune mouse model of botulinum toxin AHc vaccine. Background technique [0002] Botulinum neurotoxin (BoNT) is a neuroexotoxin produced by Clostridium botulinum in an anaerobic environment, and it is a highly toxic protein toxoid. Botulinum toxin is a typical type A-B neuroexotoxin. By inhibiting the release of acetylcholine vesicles at the neuromuscular junction, it blocks the impulse conduction at the neuromuscular junction, eventually leading to skeletal muscle paralysis and death from respiratory failure. It is the most toxic drug known. strong protein. [0003] Botulinum toxin is a double-chain structure consisting of a light chain (50 kDa) and a heavy chain (100 kDa), linked by disulfide bonds. Light chains are toxic components. The heavy chain is the toxicity receptor of nerve cells, which is not active itself, and is necessary for mediating the transmembrane transport of toxins and ent...
Claims
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IPC IPC(8): A61K39/08A61P31/04A61P39/02A61P21/00A01K67/027
CPCA61K39/08A61P31/04A61P39/02A61P21/00A01K67/027A61K2039/544A01K2267/03A01K2227/105A01K2207/20
Inventor 杨慧盈干长姣周冬生孙岩松法云智高波邱业峰焦周光郝淮杰杨文慧胡凌飞赵月娥于学东熊小路焦俊殷喆
Owner ACADEMY OF MILITARY MEDICAL SCI