Integrated Solution Of Veterinary Vaccines

I. Introduction:

Veterinary vaccines are those dedicated to animals, which belong to veterinary biological products for prevention. The integrated solution of veterinary vaccines includes customization of separation and purification of medium, selection of chromatography apparatus and chromatographic column, test of purification process and application on production line, personnel operation training and technical guidance, to provide top quality products and services.

The veterinary vaccine culture medium is composed of medium, host metabolites, host cells and other substances, and the components are complex. Traditional PEG concentration or sucrose density centrifugal centrifugation and purification only removes water and partial micromolecules, leaving biomolecules with size close to that of vaccine. After vaccination, the animal may have a serious stress response with severe side effects, which may threaten safety of aquaculture industry and bring a huge economical loss. To improve safety of animal vaccines, purification is one important means.

II. Detailed introduction:

1. Separation and purification:

Membrane filtering, sieve chromatography, ion-exchange column chromatography, affinity chromatography, etc.

1) Membrane filtering

Membrane filtering is a sieving process related to membrane pore size, during which micropores distributed on the membrane surface only allow passing of water and micromolecules to form permeate when stoste flows through the membrane surface under a certain pressure with pressure difference on both membrane sides as the drive force and membrane as the filter medium, while materials with size larger than micropore size in the stoste are remained at inlet side of membrane, forming concentrates, thus realizing separation and concentration of stoste.

2) Sieve chromatography (gel filtration chromatography)

It is one chromatographic technique which uses gel with pores of a certain size as chromatography medium (such as dextrangel, sepharose gel, polyacrylamide gel, etc.) to separate materials with different molecular weights and shapes. Due to different size and shape of various molecules, the speed spreading into gel’s pores is different, thus separation is realized by different speed of passing through chromatographic columns.

3) Ion-exchange column chromatography

Stationary phase is the chromatographic technique of ion exchanger. Solute ions to be separated in the sample exchange with ions combined at stationary phase. The affinity and binding conditions with ionized groups on ion exchanger is different for different solute ions. When eluent flows through, ions in the sample are eluted in turn based on different binding. In biochemistry and molecular biology industries, this method is usually used to separate biomacromolecules, such as protein, nuclear acid.

4) Affinity chromatography

It is a chromatography technology making use of the special and reversible affinity binding between molecules and ligands for separation. Various kinds of chromatographic separation methods can be adopted, which are designed with affinity interaction of biochemistry, chemoimmunology or anastomosis of other textures. For example, oligodeoxythymidylic acid-cellulose is used for separation and purification of mRNA, DNA- cellulose for separation of DNA-dependent DNA polymerase, agarose-antibody preparation for separation of antigen, and metal chelate column for separation of recombinant protein with string histidine-tag.

2. Extraction and concentration:

The extraction equipment shall be configured as per quality standards, process requirements and production scale of proposed product types.

1) Main equipment: extraction tank (decoction tank, percolation tank or multi-functional extraction tank), liquid storage tank, concentrator, ethanol blending tank, ethanol storage rank, settling tank, storage facility, filter, dryer, etc.

2) The volume of one extraction tank shall not be less than 1T (quantitative), and be matched with performance and capacity of supporting concentration, refining, filtering and other equipment;

3) For products adopting reflux extraction process with traditional Chinese medicine volatile oil as the preparation raw materials, its extraction facility shall be closed and equipped with a cooling system. Besides, it shall be set up with a device for separating and collecting volatile oil and an aqua aromatica tank.

4) Main extraction processes for veterinary vaccines or Chinese veterinarian

★ Extraction methods: decocting, percolation, impregnation, reflux, steam distillation, supercritical fluid extraction, semi-bionic extraction, ultrasonic extraction and so on;

★ Concentration methods: reduced pressure evaporation, atmospheric evaporation, film evaporation, multi-effect evaporation, etc.;

★ Refining methods: water extraction and alcohol precipitation (water alcohol method), alcohol extraction and water precipitation (alcohol water method), acid precipitation, adsorption and clarification, resin macroporous adsorption, salt fractionation, dialysis, etc.;

★ Clarification and filtering methods: sedimentation separation, centrifugal separation, filtering separation (reduced pressure filtering, atmospheric filtering, pressure filtering, membrane filtering), etc.

★ Drying methods: drying, reduced pressure drying, spray drying, boiling drying, freeze drying, microwave drying, infrared drying, drum drying, band drying, hygroscopic drying, etc.