Rapid extraction method of immunoglobulin Y as well as prepared anti-hand, foot and mouth disease product and application of immunoglobulin Y

Abstract

The invention discloses a rapid extraction method of immunoglobulin Y and application of the immunoglobulin Y to preparation of an anti-hand, foot and mouth disease product. The rapid extraction method comprises the following steps: laying hens are immunized with a tetravalent enterovirus vaccine of EV71, CoxA16, CoxA10 and CoxA6, and a bacterial vaccine of escherichia coli and staphylococcus aureus; eggs are separated mechanically and automatically to obtain egg yolks, and a citric acid-disodium hydrogen phosphate buffer solution is adopted to dilute and stabilize a pH value of an egg yolk solution; dry ice is added to rapidly reduce the system temperature; continuous high-speed low-temperature centrifugation is performed to remove lipid particles and obtain immunoglobulin Y supernatant;the immunoglobulin Y supernatant passes through a double-layer filter membrane of 2-6[mu]m and 0.3-0.8[mu]m sequentially, and a hollow fiber ultrafiltration membrane with molecular weight cut-off of 300kD to remove impurity protein, and finally the immunoglobulin Y supernatant passes through the hollow fiber ultrafiltration membrane with the molecular weight cut-off of 100kD to obtain a concentrated immunoglobulin Y solution; and auxiliary materials are added to prepare a spray for skin and mucosas of mouth and nasal cavity. The method has the advantages of high automation degree, low energy consumption, large output, green and environmental protection, large-scale industrial production and the like.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, especially the biotechnology of obtaining antibodies through immunization and breeding technology and preparing them into medicines or health care products, and particularly relates to a method for rapidly extracting egg yolk antibodies and an anti-hand, foot and mouth disease product prepared from the extracted egg yolk antibodies and its application. Background technique [0002] Hand, foot and mouth disease is an infectious disease commonly seen in children under the age of 5. The characteristics of the case are fever for 3-4 days, followed by vesicle symptoms on the palms and soles of the skin, oral mucosa and tongue. The disease is mainly caused by more than 20 kinds of enteroviruses, mainly Coxsackievirus A16 (Cox A16) and enterovirus 71 (EV71). At present, the disease lacks effective drugs and mainly symptomatic treatment. [0003] Yolk antibody (Immunoglobulin Y, IgY) refers to the ...

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Problems solved by technology

For example: (1) The egg yolk is collected by manual beating, which is inefficient and not suitable for large-scale use, and is easy to introduce pollution sources, increasing the risk of contamination of egg yolk antibodies; (2) Repeated freezing and thawing 1-3 times If it is applied to production operations, it will be time-consuming and laborious; (3) use organic solvents such as octanoic acid or chloroform to extract IgY, the application of organic solvents will affect the activity of IgY, and it is toxic, pollutes the environment, and is inconvenient to operate; (4) uses Ammonium sulfate salting out precipitation method, there are time-consuming operations such as centrifugation to collect precipitates and redissolving precipitation; (5) Extraction of IgY by water dilution extraction method is suitable for large-scale production and protection of IgY activity, but it prolongs the extraction time and requires Only by placing the liquid in a low temperature environment for more than 8 hours can its temperature be reduced to 0-8°C. In actual production, this production process consumes time and energy, and the efficiency of large-scale industrial production is low.

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