CN105771713A - Novel method for emulsifying trace antigen - Google Patents
Novel method for emulsifying trace antigen Download PDFInfo
- Publication number
- CN105771713A CN105771713A CN201610129305.1A CN201610129305A CN105771713A CN 105771713 A CN105771713 A CN 105771713A CN 201610129305 A CN201610129305 A CN 201610129305A CN 105771713 A CN105771713 A CN 105771713A
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- CN
- China
- Prior art keywords
- antigen
- trace
- emulsifying
- adjuvant
- novel method
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- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
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Classifications
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01F—MIXING, e.g. DISSOLVING, EMULSIFYING OR DISPERSING
- B01F23/00—Mixing according to the phases to be mixed, e.g. dispersing or emulsifying
- B01F23/40—Mixing liquids with liquids; Emulsifying
- B01F23/405—Methods of mixing liquids with liquids
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01F—MIXING, e.g. DISSOLVING, EMULSIFYING OR DISPERSING
- B01F33/00—Other mixers; Mixing plants; Combinations of mixers
- B01F33/25—Mixers with loose mixing elements, e.g. loose balls in a receptacle
- B01F33/251—Mixers with loose mixing elements, e.g. loose balls in a receptacle using balls as loose mixing element
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- Chemical & Material Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
Abstract
The invention provides a novel method for emulsifying a trace antigen.The antigen and a Freund's adjuvant at the same volume are mixed in a sterile 1.5ml EP tube, sterile steel balls are added in proportion, other EP tubes are used for balancing, a tube opening is sealed through a sealing film, a machine is started at the rotating speed of 1,500 revolutions per min, the tube is taken out 1 min later and placed in an ice bath for 1 min, the operation is repeated four times, 1,000 g of the emulsified antigen is centrifuged for 1 min, and the mixture is not layered.The novel method has the advantages that a trace tissue crusher is applied to emulsification of the trace antigen, in combination with the steel balls of different specifications, multiple different trace antigens can be effectively emulsified at the same time, the problems are effectively solved, large-scale preparation of the different antigens is completely achieved, and good application potential is achieved.
Description
Technical field
Invention belongs to immunological technique field, it is provided that the new method of a kind of trace antigen emulsifying, is the improvement to existing trace antigen Yu immunological adjuvant emulsification method.
Background technology
The immunoreation of inducement efficient is the great majority purposes based on the active immunity of antigen inoculation.For improving the immune effect of antigen, generally effectively being mixed with adjuvant by antigen, such mixture can extend antigen release time after being expelled to animal thus providing lasting antigenic stimulus, to obtain good immune effect.Although the antigen used is different, but freund adjuvant is more common immunological adjuvant all the time.Be no matter Fu Shi completely or Freund's incomplete adjuvant, existence because of a large amount of lanolines, its viscosity is made extremely to increase, when water-soluble antigen joins in this type of adjuvant, generally adopt syringe pull method repeatedly, mortar polishing, mechanical mixing method etc. to act on through the long period and just can make water in oil antigen-adjuvant mixture.Syringe pull method is relatively time-consuming, effort, also makes the good antigen of part emulsifying be stranded in emulsifying injection simultaneously, is not suitable for the emulsion process of preciousness, trace antigen；Mortar polishing and mechanical mixing method cause antigen to be polluted by inoculating microbe because of sealing problem, and cause nonspecific immune reaction.Therefore seek for trace antigen emulsification method very necessary.Ultrasonic emulsification is suitable for the emulsifying of trace antigen, it largely avoid antigen by the pollution of exogenous microbial, but ultrasound wave can cause the structural change of particulate antigen, and then have influence on its immunogenicity, therefore operation sequence needs system to grope to carry out realizing sequencing, is not suitable for the emulsifying of trace, precious antigen.
Summary of the invention
The invention provides the new method of a kind of trace antigen emulsifying, mix with freund adjuvant etc. for trace antigen and prepare water-in-oil type immunity mixture, solve the deficiency that existing antigen emulsifying technology exists.
Antigen involved in the present invention is the protein matter with inducing immunogenic.
Trace involved in the present invention refers to that volume range is between 0.01 ~ 1000 microlitre.
Adjuvant involved in the present invention is often referred to Freund's complete adjuvant and freund 's incomplete adjuvant, also includes other oil adjuvants that can produce water-in-oil type immunity mixture.
The instrument BulletBlender that the present invention uses is be a novelty, the broken instrument of histiocyte quick, high flux, high stability.Its rotary head impacts test tube with the frequency of several thousand times per minute, when adding the high-wearing feature grinding bead of different size in the sample, can cell or tissue sample be crushed and homogenizing thoroughly.
The plastics EP centrifuge tube that mixing container is 1.5-2.0 milliliter used in the present invention.
The new method of a kind of trace antigen emulsifying of the present invention, comprises the following steps:
The antigen of suitable concentration and isopyknic freund adjuvant are mixed in 1.5 milliliters of EP pipes of sterilizing, add the aseptic steel ball 4 ~ 8 that diameter is 0.5 ~ 2.0mm, carry out trim with other EP pipe simultaneously, seal the mouth of pipe with sealed membrane, start machine, 1500 revs/min, take out after 1 minute, be placed in ice bath 1 minute, 4 times so repeatedly, being centrifuged 1 minute with 1000g by antigen good for emulsifying, mixture is not stratified.
The positive effect of the present invention is in that:Microcomponent's disintegrating machine is applied to the emulsifying of trace antigen by the present invention; in conjunction with the steel ball of different model, can effectively realize emulsifying while multiple different trace antigen, not only efficiently solve the problems referred to above; and prepared by the scale fully achieving not synantigen, there is good application potential.
Detailed description of the invention
For the ease of understanding the present invention, especially exemplified by following example.Its effect is understood to be the explaination to the present invention but not any type of restriction to the present invention.
Embodiment 1
The preparation of rabies virus immunity mixture
By the inactivation rabies virus purified 0.2 milliliter of 1000 mcg/ml with isopyknic Freund's complete adjuvant after 1.5 milliliters of EP pipes mix, add the steel ball 6 of diameter 2mm, seal the mouth of pipe, start machine, 1500 revs/min, take out after 1 minute, in ice bath 1 minute, repeating 4 times, virus antigen and freund adjuvant are after 1000g is centrifugal 1 minute, and layering does not occur in mixture.
Embodiment 2
The preparation of bovine serum albumin immunity mixture
The bovine serum albumin of 1 milligram is dissolved in 1 ml physiological saline, take 0.1 milliliter to be added in 1.5 milliliters of EP pipes, add isopyknic Freund's complete adjuvant, add the steel ball 4 of diameter 2mm, other steps press embodiment 1, and the mixture that emulsifying is good does not find layering when centrifugal.
Above institute embodiment employs particulate antigen and proteantigen two class, and the adjuvant used is Freund's complete adjuvant, owing to the physical behavior of freund 's incomplete adjuvant is identical with Freund's complete adjuvant, therefore as selected freund 's incomplete adjuvant also can realize identical emulsifying effectiveness.The use of two class antigens in the present invention simultaneously, illustrates to present invention can apply to macromole and small molecule antigens.
Claims (1)
1. a new method for trace antigen emulsifying, comprises the following steps:
The antigen of suitable concentration and isopyknic freund adjuvant are mixed in 1.5 milliliters of EP pipes of sterilizing, add the aseptic steel ball 4 ~ 8 that diameter is 0.5 ~ 2.0mm, carry out trim with other EP pipe simultaneously, seal the mouth of pipe with sealed membrane, start machine, 1500 revs/min, take out after 1 minute, be placed in ice bath 1 minute, 4 times so repeatedly, being centrifuged 1 minute with 1000g by antigen good for emulsifying, mixture is not stratified.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610129305.1A CN105771713A (en) | 2016-03-08 | 2016-03-08 | Novel method for emulsifying trace antigen |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610129305.1A CN105771713A (en) | 2016-03-08 | 2016-03-08 | Novel method for emulsifying trace antigen |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN105771713A true CN105771713A (en) | 2016-07-20 |
Family
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Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201610129305.1A Pending CN105771713A (en) | 2016-03-08 | 2016-03-08 | Novel method for emulsifying trace antigen |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN105771713A (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110665379A (en) * | 2019-10-08 | 2020-01-10 | 浙江农林大学 | Water-in-oil type emulsification method aiming at vaccine antigen |
| CN115253736A (en) * | 2022-07-21 | 2022-11-01 | 苏州逻晟生物医药有限公司 | Novel antigen emulsification method |
-
2016
- 2016-03-08 CN CN201610129305.1A patent/CN105771713A/en active Pending
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110665379A (en) * | 2019-10-08 | 2020-01-10 | 浙江农林大学 | Water-in-oil type emulsification method aiming at vaccine antigen |
| CN115253736A (en) * | 2022-07-21 | 2022-11-01 | 苏州逻晟生物医药有限公司 | Novel antigen emulsification method |
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| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| WD01 | Invention patent application deemed withdrawn after publication | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20160720 |