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Chicken yoke antibody resisting Bengalese cobra venom and its preparation

A technology of egg yolk antibody and snake venomous chicken, which is applied in the field of immunoglobulin and its preparation, can solve the problems that there is no anti-Bengal cobra, etc., and achieve good therapeutic effect, wide application range and high yield

Inactive Publication Date: 2006-11-08
广州医学院
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] At present, scholars at home and abroad have prepared several kinds of IgY such as anti-Brazilian anti-rattlesnake venom, anti-snake venom, anti-viper venom, anti-viper venom, anti-king cobra venom and anti-Chinese cobra, etc., which have obvious affinity and specificity. Higher than horse-derived antiserum, but so far there is no report on the preparation and application of anti-Bengal cobra (Naja kauthia) venomous chicken egg yolk antibody

Method used

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  • Chicken yoke antibody resisting Bengalese cobra venom and its preparation

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Embodiment 1

[0023] (1) Preparation of Bengal cobra venom antigen: adult Bengal cobra, body weight: 1.5~2.0kg, using the venom extractor disclosed in the patent No. 200420046279.9, the venom is diluted with distilled water in a ratio of 1:4, centrifugal, vacuum, low temperature Dried to lyophilized powder. Dissolve 50 mg of snake venom freeze-dried powder in 5 ml of 0.1M PBS, pH 6.8, add 0.5 ml of 0.25% glutaraldehyde and place it at 30°C for 2 hours, use Sephadex G-25 chromatography to remove salt, and determine snake venom LD 50 .

[0024] (2) Adjuvant emulsification and chicken immunization: take 22-week-old Laihang hens (body weight: 1.0±0.1kg), fully emulsify Bengal cobra venom and Freund’s adjuvant at a ratio of 1:1 (V / V), Subcutaneous and intramuscular multi-site injection through chicken wings, chest, abdomen and back. The snake venom was emulsified with Freund's complete adjuvant for the first immunization, and the snake venom was emulsified with Freund's incomplete adjuvant for...

Embodiment 2

[0032] (1) Bengal cobra venom antigen preparation: adult Bengal cobra, body weight: 1.5~2.0kg, using the venom taking device disclosed in the patent No. 200420046279.9, the venom is diluted with distilled water in a ratio of 1:4, centrifugal, vacuum, low temperature Dried to lyophilized powder. Dissolve 50 mg of snake venom freeze-dried powder in 5 ml of 0.1M PBS, pH 6.8, add 0.5 ml of 0.25% glutaraldehyde at 30°C for 2 hours, use Sephadex G-25 chromatography to remove salt, and determine snake venom LD 50 .

[0033] (2) Adjuvant emulsification and chicken immunization: take 22-week-old Laihang hens (body weight: 1.0±0.1kg), fully emulsify Bengal cobra venom and Freund’s adjuvant at a ratio of 1:1 (V / V), Subcutaneous and intramuscular multi-site injection through chicken wings, chest, abdomen and back. The snake venom was emulsified with Freund's complete adjuvant for the first immunization, and the snake venom was emulsified with Freund's incomplete adjuvant for subsequent ...

Embodiment 3

[0041] (1) Bengal cobra venom antigen preparation: adult Bengal cobra, body weight: 1.5~2.0kg, using the venom taking device disclosed in the patent No. 200420046279.9, the venom is diluted with distilled water in a ratio of 1:4, centrifugal, vacuum, low temperature Dried to lyophilized powder. Dissolve 50 mg of snake venom freeze-dried powder in 5 ml of 0.1M PBS, pH 6.8, add 0.5 ml of 0.25% glutaraldehyde at 30°C for 2 hours, use Sephadex G-25 chromatography to remove salt, and determine snake venom LD 50 .

[0042] (2) Adjuvant emulsification and chicken immunization: take 22-week-old Laihang hens (body weight: 1.0±0.1kg), fully emulsify Bengal cobra venom and Freund’s adjuvant at a ratio of 1:1 (V / V), Subcutaneous and intramuscular multi-site injection through chicken wings, chest, abdomen and back. The snake venom was emulsified with Freund's complete adjuvant for the first immunization, and the snake venom was emulsified with Freund's incomplete adjuvant for subsequent ...

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Abstract

The present invention is chicken yoke antibody (IgY) resisting Bengalese cobra venom and its preparation process. The preparation process includes water dilution to extract coarse IgY from chicken yoke, salting out IgY with ammonium sulfate, dissolving the precipitate in PBS, ultrafiltering to desalt, concentrating, anion exchanging chromatographic separation for further purifying IgY, ultrafiltering to desalt, concentrating, and freeze drying. The IgY exhibits single tape of molecular weight 200 KD in non-reductant SDS-PAGE and two tapes of molecular weight 65 KD and 35 KD separately in reductant SDS-PAGE, and has at least 13 dyeing strips of molecular weight 100-0.7 KD on PVDF film for Western Blot detection. The chicken yoke antibody (IgY) resisting Bengalese cobra venom may be used in preparing medicine and health food for preventing and / or treating snake bite, and in preparing reagent for detecting relevant snake venom.

Description

technical field [0001] The invention relates to an immunoglobulin in the biological field and a preparation method thereof, in particular to an anti-Bengal cobra (Naja kaouthia) venomous chicken egg yolk antibody and a preparation thereof. Background technique [0002] Venomous snake bites are sudden, severe, progress rapidly and have a high mortality rate. Applying specific antivenom serum as soon as possible is the most effective way to treat snake bites. Among the highly venomous snakes in southern my country, there are mainly two species of cobras belonging to the subfamily Cobra, namely the Chinese cobra (Naja atra) and the Bengal cobra (Naja kaouthia). The Chinese cobra, also known as the Zhoushan cobra, is characterized by the obvious white speckled markings on the back of its nape. The first choice for the bite of this snake is the refined antivenom of the Chinese cobra. Bengal cobra, also known as Thai cobra, has only a single "glass"-shaped spot on the nape. It wa...

Claims

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Application Information

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IPC IPC(8): C07K16/02A61K39/395A61P39/02C07K1/14
Inventor 孔天翰刘四红董伟华黄热平黄劭覃媛
Owner 广州医学院
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