Anti-blood-cycle-venin egg yolk antibody and application thereof
A technology of egg yolk antibody and snake venom chicken, which is applied in the field of blood circulation type snake venom chicken egg yolk antibody, can solve the problems limited to the classification and identification of venomous snakes and basic theoretical research, and achieve small allergic adverse reactions, high yield and good stability Effect
Active Publication Date: 2014-05-14
ZHEJIANG CHINESE MEDICAL UNIVERSITY
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Problems solved by technology
Retrieval of related literature has found that many monoclonal antibodies related to snake venom protein have been successfully prepared (Yin Huiqiong. Preparation of anti-venom ScFv antibody using phage display library technology. Yunnan University, 2004 master thesis.; Li Lihong et al. Preparation of monoclonal antibody against king cobra venom . Cellular and Molecular Immunology Journal, 2009,25(3):236-238. etc.), but most of them are limited to the classification and identification of venomous snakes and basic theoretical research; this is most likely due to the diversity of venomous components (Jia Yan et al. Toxic components of snake venom and their application research. Snake Zhi, 2004,16(2):23-32.), so almost no monoclonal antibody can completely neutralize the toxicity of a certain snake venom
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Embodiment 1
[0027] Example 1: Preparation of novel anti-circulatory snake venom chicken egg yolk antibody based on snake venom zinc ion metalloprotease activity motif
[0028] 1. Synthesis and coupling of peptides
[0029] Through the comparative analysis of the amino acid sequences of the catalytic domains of various snake venom metalloproteases (such as Acutolysin A, Acutolysin F, Acutolysin E, Aculysin 2, Metalloprotease H1, Metalloprotease H2, Metalloprotease H3, Metalloprotease H4 and Metalloprotease H5), the synthesis of the following peptides was determined Sequence (M15): CAHEMAHNLGVRHDE, and submitted to Hangzhou Jucheng Biotechnology Co., Ltd. Synthetic peptide, HPLC and MS identification results are as follows figure 1 , 2 shown. Then the polypeptide was coupled to keyhole limpet hemocyanin (KLH, purchased from sigma) by the carbodiimide method to form the antigenic peptide complex KLH-M15.
[0030] 2. Hens are immunized
[0031] The above antigen peptide complex KLH-M15 ...
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Abstract
The invention discloses an anti-blood-cycle-venin egg yolk antibody. The antibody is prepared in the following steps of: synthesizing venin zinc ion metalloproteinase active structure domain polypeptide based on a blood-cycle type venin zinc ion metalloproteinase active structure domain sequence; coupling, cohering or mixing the polypeptide with the carrier to prepare an antigen peptide compound; injecting the antigen peptide compound in a hen body for immunization, and continuously collecting eggs; extracting an anti-venin egg yolk antibody from the collected eggs to obtain the anti-blood-cycle-venin egg yolk antibody. The anti-blood-cycle-venin egg yolk antibody is high in yield and low in cost; and the prepared antibody is high in safety and stability and low in anaphylactic reaction. The anti-blood-cycle-venin egg yolk antibody can be prepared into oral medicine or injection, can be used for treating blood-cycle-venin bite wound, and is particularly suitable for treating bite wound by a long-noded pit viper or a bobtail hagworm.
Description
(1) Technical field [0001] The invention relates to a blood circulation type snake venom chicken egg yolk antibody (Immunoglobulin-yolk, IgY), a preparation method and an application. Specifically, it is a preparation method for preparing an anti-blood circulation type snake venom chicken egg yolk antibody based on the immunization of the blood circulation type snake venom zinc ion metalloprotease active motif polypeptide. (2) Background technology [0002] For a long time, poisonous snake bites have brought serious harm to people's production and life. According to the figures released by the World Health Organization website: more than 2.5 million people are bitten by snakes in the world every year, resulting in 125,000 deaths. In my country, more than 100,000 patients are bitten by poisonous snakes every year, of which about 73% are young and middle-aged, the mortality rate is 5% to 10%, and the disabled and incapacitated by snakebite account for 25% to 30%. It can be se...
Claims
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IPC IPC(8): C07K16/40C07K16/02A61K39/395A61P39/02
Inventor 范永升蒋福升丁志山刘翔宇吕迪潘平毛建洋
Owner ZHEJIANG CHINESE MEDICAL UNIVERSITY