CN100510748C - Method of evaluating performance of activation gas deactivating antigenic substance and apparatus for generating processed antigenic substance used as evaluation sample of the evaluating method - Google Patents

Method of evaluating performance of activation gas deactivating antigenic substance and apparatus for generating processed antigenic substance used as evaluation sample of the evaluating method Download PDF

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CN100510748C
CN100510748C CNB2004800104221A CN200480010422A CN100510748C CN 100510748 C CN100510748 C CN 100510748C CN B2004800104221 A CNB2004800104221 A CN B2004800104221A CN 200480010422 A CN200480010422 A CN 200480010422A CN 100510748 C CN100510748 C CN 100510748C
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antigenicity substance
aforementioned
activated gas
antigenicity
substance
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CN1777808A (en
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西川和男
野岛秀雄
米田哲也
小埜和久
重田征子
大下昌利
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Sharp Corp
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Sharp Corp
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Abstract

It is intended to provide method of evaluating the performance of an activated gas of inactivating an antigenic substance which comprises the step (S101) of reacting the antigenic substance with the activated gas to thereby give a treated antigenic substance; and the step (S103) of reacting an antibody against the antigenic substance with the treated antigenic substance and measuring the binding activity of the treated antigenic substance to the antibody. According to this method, the performances of various activated gases of inactivating various antigenic substances can be accurately and conveniently evaluated.

Description

Activated gas makes the method for evaluating performance of antigenicity substance inactivation, as the generating apparatus of the processed antigen material of the evaluation sample of this evaluation method
Technical field
The present invention relates to the method for evaluating performance that activated gas makes the antigenicity substance inactivation.More detailed content is to the present invention relates to make the antigenicity substance inactivation estimate the method for the performance of activated gas thus by as making mammal produce the antigenicity substance and the activated gas reaction of the material of allergic effect reaction.
The invention still further relates to the generating apparatus of the antigenicity substance of activated gas processing in addition.More detailed content is the generating apparatus that the present invention relates to as the processed antigen material of the activated gas performance evaluation sample that makes the antigenicity substance inactivation, and wherein this device possesses container.
Background technology
In recent years, variation along with living environment, press for to remove and cause the mammiferous pollinosis that comprises the people, airborne harmful suspended materials such as pollen, tick, tick ight soil and chamber dirt of allergic disease reasons such as asthma, the congenital allergy of skin, conjunctivitis are built healthy, comfortable life.
In order to satisfy this demand, developed and effectively to have removed the antigenicity substance (anaphylactogen) that forms above-mentioned allergic disease reason, and used the air purifier (for example opening flat 8-173843 communique) of various filtrators and dust suction mode with reference to the spy.
But, in such conditioner, owing to be the air that sucks in the atmosphere, method by filtrator absorption or filtering noxious suspended material, so because long-term use, must to change the maintaining of filtrator etc., and because the characteristic imperfection of filtrator, so sometimes can not get satisfactory performance.
And in such conditioner, for example, when being purpose with capture pollen, pollen is under the antigen protein existence that forms the pollinosis reason, become trapped in the catcher/filter with physical method left behind.This pollen that is captured through physical method is easy to from catcher/filter come off, so when entry into service, when shutting down or when changing filtrator, exist the problem that might cause that the pollen that is captured disperses once more.In addition, promptly allow to a pollen and self trap in the catcher/filter, and the particle diameter antigen protein littler than pollen might pass through catcher/filter, therefore the problem that also can cause fundamentally to remove antigenicity substance.
On the basis of various filtrators, developed the pollen treating apparatus (for example opening flat 7-807 communique) that makes the antigenicity substance sex change by heat treated again simultaneously with reference to the spy.
But in this conditioner, heat treated need consume a large amount of energy, and existing to increase family's electricity charge expenditure, can bring dysgenic problem to earth environment simultaneously.And when using this conditioner in summer or in the high temperature area, indoor temperature can obviously rise, and exists the problem that the people is felt under the weather.Therefore also existing can not be with its problem of using with thermantidote.
On the basis of various filtrators, developed again by carrying out the ultraviolet ray irradiation, make the device (for example opening flat 6-154298 communique) of Chinese fir pollinosis antigen inactivation with reference to the spy.
But in this conditioner, carrying out the ultraviolet ray irradiation needs to consume a large amount of energy, and existing to increase family's electricity charge expenditure, can bring dysgenic problem to earth environment simultaneously.And, during the antibody valence of the sample that causes by Chinese fir pollen, must carry out minimum intensity also at 1.3mW/cm according to described in the above-mentioned document 2Or its above, 50 seconds or its above ultraviolet ray irradiation.Make the ability of Chinese fir pollinosis antigen inactivation low, therefore it is a kind of technology with Practical significance hardly.
On the basis of various filtrators, developed again by carrying out ultraviolet ray irradiation, ozoniferous air purifier (for example opening the 2000-111106 communique) with reference to the spy.
But in this conditioner, carrying out the ultraviolet ray irradiation needs to consume a large amount of energy, and existing to increase family's electricity charge expenditure, can bring dysgenic problem to earth environment simultaneously.And ozone can be discharged in the surrounding atmosphere, under certain conditions, can cause harmful effect to the mammal live body that comprises the people.
In all these conditioners, not having can be according to the antigenicity substance kind of the allergic effect reaction that has individual difference because of different people, and antigenicity substance is carried out the problem of different disposal, also all solves fully.So, variously remove method or different these problems of effect of its method for deactivating also do not solved corresponding to different types of antigenicity substance.
According to above-mentioned present situation, problem of the present invention is to provide the method that the performance of various antigenicity substance inactivations is estimated to various activated gas, this method is for realizing that a kind of conditioner is necessary, described conditioner can effectively be removed antigenicity substance and/or make its inactivation by activated gas, and the kind of described activated gas and/or consumption are corresponding with the kind and/or the consumption of the antigenicity substance that has difference because of individuality is different.
Another problem of the present invention provides the generating apparatus of processed antigen material, and this installs in above-mentioned evaluation method, can evenly and easily generate as estimating processed antigen material sample, that handle by activated gas.
Summary of the invention
In order to solve above-mentioned problem, the inventor makes great efforts to study the evaluation method of groping to make for establishment the activated gas performance of antigenicity substance inactivation.
The inventor finds as a result, by in container, scattering antigenicity substance, make and contain the solution that scatters antigenicity substance to some extent and be suspended under the state in the container, antigenicity substance and activated gas are reacted, by this reaction, the antigenicity substance of can easy acquisition uniform, activated gas processing.
And the inventor also finds, by using this processed antigen material, can accurately and easily estimate the performance that activated gas makes the antigenicity substance inactivation.
Activated gas just of the present invention comprises the evaluation method of the performance of antigenicity substance inactivation and antigenicity substance and activated gas is reacted, obtain the step of processed antigen material; With antibody and this processed antigen material at this antigenicity substance are reacted, measure the step of this processed antigen material with respect to this antibody binding activity.
Activated gas of the present invention makes the method for evaluating performance of antigenicity substance inactivation, is method as described below, and it comprises reacts antigenicity substance and activated gas, obtains the step of processed antigen material; Antibody and this processed antigen material at this antigenicity substance are reacted, measure the step in conjunction with activity of this processed antigen material with respect to this antibody; With with this processed antigen material in conjunction with active and combine the step that activity compare of this antigenicity substance with respect to this antibody.
Wherein, the step that obtains this processed antigen material preferably includes the step that suspend skyborne this antigenicity substance and this activated gas are reacted.
Simultaneously, in the step of carrying out this reaction, be preferably included in the container and scatter the step that contains this antigenicity substance solution, make the solution that contains this antigenicity substance that is scattered be suspended in the step in this container and in this container, import the step of this activated gas.
In obtaining the step of this processed antigen material, preferably make this antigenicity substance skyborne step that suspends by this antigenicity substance being applied vibration and/or impacting.
In this suspension step, preferably include the step that this antigenicity substance is arranged on the step on the sample bench with flexibility and this sample bench is applied vibration and/or impacts.
The suspension step here be aforementioned antigenicity substance be arranged on the sample bench that one or more that be selected from bed clothes, woollen blanket, cushion, pillow, mat, sponge, cloth, paper, styrenic foams have flexibility step and by aforementioned sample bench being beaten and/or vibrates, the step that aforementioned sample bench is applied vibration and/or impacts.
The step that this obtains the processed antigen material preferably includes the step that makes this antigenicity substance and contain one or more gas reactions that are selected from the gas that contains positive ion, the gas that contains negative ion, the gas that contains free radical, ozone gas, nitric acid gas.
This obtains the step of processed antigen material, preferably include and make a kind or its above material and the activated gas reaction that is selected from antigenicity substance contained in Chinese fir pollen and/or the tick dust, Chinese fir pollen, tick dust, obtain the step of processed antigen material.
This determination step preferably includes by ELISA method and/or ELISA and suppresses method, and antibody and this processed antigen material at this antigenicity substance are reacted, and measures the step of this processed antigen material for this antibody binding activity.
This determination step in addition, also preferably include to the generation of possessing except that the people and carry out intradermal reaction test and/or conjunctival reaction test at the animal of the cell of the antibody of this antigenicity substance, make this antibody and this processed antigen substance reaction, measure the step of this processed antigen material for this antibody binding activity.
Generating apparatus as the employed processed antigen material of performance evaluation sample of the antigenicity substance inactivation of activated gas of the present invention possesses container is arranged, scatters the device of antigenicity substance and the device of this activated gas takes place or imports in this container in this container.
The present invention also provides as estimating the generating apparatus of processed antigen material that activated gas makes the performance sample of antigenicity substance inactivation, and this device possesses container, enclose the device of antigenicity substance in this container and the device of this activated gas takes place or imports in this container.
In the generating apparatus of above-mentioned any one processed antigen material of the present invention, preferred container contains the partially transparent material or all is transparent material.
The simple declaration of accompanying drawing
Fig. 1 is the outline flowchart that expression activated gas of the present invention makes the method for evaluating performance of antigenicity substance inactivation.
Fig. 2 enumerates an example explanation as estimating the sketch map of processed antigen material generating apparatus of sample that activated gas of the present invention makes the performance of antigenicity substance inactivation.
Fig. 3 enumerates another example explanation as estimating the sketch map of processed antigen material generating apparatus of sample that activated gas of the present invention makes the performance of antigenicity substance inactivation.
Fig. 4 enumerates another example explanation as estimating the sketch map of processed antigen material generating apparatus of sample that activated gas of the present invention makes the performance of antigenicity substance inactivation.
Fig. 5 enumerates another example explanation as estimating the sketch map of processed antigen material generating apparatus of sample that activated gas of the present invention makes the performance of antigenicity substance inactivation.
Fig. 6 enumerates another example explanation as estimating the sketch map of processed antigen material generating apparatus of sample that activated gas of the present invention makes the performance of antigenicity substance inactivation.
Fig. 7 is the sketch map that illustrates used ion generating device structure among the present invention
Fig. 8 A and 8B are that expression is by the positive ion of ion generating device generation and the mass spectrum of negative ion.
Fig. 9 A and 9B be expression when the Chinese fir antigenicity substance being handled and when processing by the gas that contains the both positive and negative ion, the chart of antigenicity substance and pollinosis patient's 19～40 SERUM IgE antibody allergic effect reaction relation.
Figure 10 A and 10B be expression when the Chinese fir antigenicity substance being handled and when processing by the gas that contains the both positive and negative ion, the chart of antigenicity substance and pollinosis patient's 41～60 SERUM IgE antibody allergic effect reaction relation.
Figure 11 be expression when the Chinese fir antigenicity substance being handled and when processing by the gas that contains the both positive and negative ion, the chart of Cry j 1 and Cry j 2 and its monoclonal antibody reactive sexual intercourse.
Figure 12 be expression by enzyme linked immunological absorption inhibition method, when the Chinese fir antigenicity substance being handled and handling with the gas that contains the both positive and negative ion, the chart of the reactive relation of antigenicity substance and pollinosis patients serum IgE antibody allergic effect.
Figure 13 is the chart of various concentration with the reaction inactivation rate relation of the antigenicity substance that derives from Chinese fir pollen of both positive and negative ion in the expression activated gas.
Figure 14 lifts the sketch map that an example explanation is used to implement make the device of antigenicity substance method for deactivating, is to have the device that makes the facility that ozone concentration reduces.
Figure 15 is that expression is to antigenicity substance (tick antigenicity substance) when carrying out ion processing and when processing, with the chart of the SERUM IgE antibody allergic effect reaction relation of tick autopath a～r.
Figure 16 is the sketch map that expression enforcement makes the device of antigenicity substance method for deactivating, and it is the device that possesses fan blower and reclaim filtrator.
Figure 17 is the sketch map that expression implements to make the device of antigenicity substance method for deactivating, its be possess fan blower and returnable device.
Figure 18 is (3000/cm of space average concentration that are illustrated in the both positive and negative ion 3) under, by enzyme linked immunological absorption inhibition method, when the tick dust is carried out ion processing and when processing, the reactive chart that concerns of antigenicity substance and tick autopath's SERUM IgE antibody allergic effect.
Figure 19 is (10000/cm of space average concentration that are illustrated in the both positive and negative ion 3) under, by enzyme linked immunological absorption inhibition method, when the tick dust is carried out ion processing and when not carrying out ion processing, the reactive chart that concerns of antigenicity substance and tick autopath's SERUM IgE antibody allergic effect.
Implement preferred plan of the present invention
Below be described in more detail situation of the present invention with embodiment.
＜antigenicity substance 〉
In this instructions, biologies such as pollen class that so-called antigenicity substance is Chinese fir, cypress tree, pig dish etc. and tick, the material that is comprised in the indoor suspension of the ight soil of biologies such as tick or chamber dirt etc. etc., by acting on the mammal live body that comprises the people, the allergic effect that produces one of antigen-antibody reaction reacts and the material of induced allergy disease.
The material that this antigenicity substance normally is made of protein or glycoprotein, but in this manual, to its shape or size, not special the qualification, usually can think to comprise that these protein and glycoprotein are from the material as molecular shape, or the material of their gathering formation shapes of particle, or the antibody response position is material (also can be referred to as antigenic determinant or epi-position) of its molecularity material part or the like.
Above-mentioned antigenicity substance can be the antigenicity substance (Chinese fir antigenicity substance) that is comprised in Chinese fir pollen self or the Chinese fir pollen.Above-mentioned antigenicity substance can also be an antigenicity substance (tick antigenicity substance) contained in tick dust self or the tick dust.
Antigenicity substance with the formation Chinese fir pollinosis cause of disease is an example, in antigenicity substance, except and well-known Cry j 1 protein and Cry j 2 protein as the cause of disease material of Chinese fir pollinosis, the epi-position that also comprises Cry j 1 protein and Cry j 2 protein, also comprise the shot-like particle (Ubisch body and orbicle are otherwise known as) in the Chinese fir pollen that contains a large amount of Cry j 1 protein and Cry j 2 protein, also comprise Chinese fir pollen self.
The tick antigenicity substance is the material that tick is comprised in health, but in general living environment, compares with tick self, is to be had problems by material contained in the tick dust under the more susceptible condition.Here said tick dust is meant the food and the excreta of the part that contains tick self and tick corpse and tick health, tick and takes off the particulate of sloughing off crust and ovum etc.Antigenicity substance among the present invention also comprises such tick dust.
＜antibody response position 〉
In this instructions, so-called antibody response position is meant privileged site contained in the antigenicity substance, the position of expression and antibodies.When usually sex change takes place or is damaged (decompositions) in this antibody response position, antigenicity substance just can not with antibodies, so can suppress allergic effect and react.
＜activated gas 〉
In this instructions, so-called activated gas is meant the gas that can produce certain chemical reaction and/or physical action to antigenicity substance.With regard to the object lesson of activated gas, be not particularly limited, can enumerate the gas that contains positive ion, contain the gas of negative ion, contain simultaneously negative ions gas, contain ozone gas, contain nitric acid gas gas, contain gas of free radical or the like.In addition, in activated gas, estimate to also have the gas of various compositions at antigenicity substance.But, can adopt activated gas of the present invention described later to make the evaluation method of the performance of antigenicity substance inactivation go to find for these activated gas.
In addition as described later, the gas that contains negative ions simultaneously acts on antigenicity substance as activated gas, has the function that makes this antigenicity substance inactivation, this is not well-known phenomenon, but the inventor adopts activated gas of the present invention to make the evaluation method of the performance of antigenicity substance inactivation, is the phenomenon of finding first.
＜antigenicity substance inactivation 〉
In this instructions, the meaning of so-called antigenicity substance inactivation be meant eliminate or reduce antigenicity substance as the antigenicity substance activity.Just eliminate or reduce the ability of antigenicity substance and antibody response.
Wherein, the inventor thinks that the mechanism of the antigenicity substance inactivation that caused by activated gas is to attack the protein, particularly its antibody response position that constitutes antigenicity substance by this activated gas, makes this protein denaturation or destruction and makes the antigenicity substance inactivation.
As described later, adopt activated gas of the present invention to make the evaluation method of performance of antigenicity substance inactivation and the phenomenon found first by the inventor, make the gas that contains negative ions simultaneously act on antigenicity substance, have the function that makes this antigenicity substance inactivation as activated gas.This deactivation function realizes by making positive ion and negative ion act on antigenicity substance.
Though never knew in the past, if but according to the inventor's viewpoint, by using the gas contain positive ion and negative ion simultaneously, compare with using the gas that contains the gas of positive ion or contain negative ion respectively separately, can bring into play special inactivation effect for antigenicity substance.Can think if according to the inventor's viewpoint, use these to have the gas of negative ions simultaneously, by chemical reaction described later, produce active substance, this active substance is for the protein that constitutes antigenicity substance, particularly its antibody response position is attacked, and makes this protein denaturation or destruction (decomposition), and makes the antigenicity substance inactivation.
Just in this manual, what is called makes the antigenicity substance inactivation, can be defined as in more detail by above-mentioned makes antigenicity substance sex change or destruction (decomposition) not only can eliminate antigenicity substance, also comprise the amount that reduces this antigenicity substance of unit volume in the atmosphere gas, or reduce the antibody response position and the antibody response of this antigenicity substance.
Wherein measure the method (or define method) of antigenicity substance reaction inactivation rate (or remaining activity), have a variety ofly, can select proper method according to the kind of antigenicity substance kind and activated gas.With regard to this class assay method, not special the qualification is as using enzyme linked immunological absorption inhibition method.According to the method, measure antigenicity substance that activated gas processing crosses show 50% suppress concentration the time, compare with the concentration that the antigenicity substance of handling without activated gas 50% suppresses, if its inhibition concentration of 50% is when 5 times or its are above, then remaining activity is 20% (just reacting inactivation rate is 80%).
Judge simultaneously which kind of degree the reaction inactivation rate reaches actually, when just having the deactivation with respect to antigenicity substance in the activated gas, the kind that look activated gas species and antigenicity substance is different and different, can judge according to appropriate threshold value.For example,, be to use the gas that contains negative ions during, can use the antigenicity substance that derives from Chinese fir pollen as antigenicity substance as activated gas though be not particularly limited.
＜activated gas makes the evaluation method of the performance of antigenicity substance inactivation 〉
Fig. 1 is the outline flowchart that expression activated gas of the present invention makes the method for evaluating performance of antigenicity substance inactivation.
It is the evaluation method that possesses following steps substantially that activated gas of the present invention makes the evaluation method of the performance of antigenicity substance inactivation: make the reaction of antigenicity substance and activated gas, obtain the step (S101) of processed antigen material; Antibody and this processed antigen material at this antigenicity substance are reacted, measure the step (S103) of this processed antigen material for this antibody binding activity.Make in the evaluation method of performance of antigenicity substance inactivation at activated gas of the present invention, shown in the process flow diagram of Fig. 1, continue the step (S101) that obtains above-mentioned processed antigen material with after measuring the step (S103) in conjunction with activity of processed antigen material, also preferably have the active step (S105) that compares for described antibody binding activity with this antigenicity substance of the combination of this processed antigen material.
By the evaluation method that adopts this and control sample to compare, has the advantage of can be accurate and easy and can carrying out quantitative evaluation to the deactivation of this antigenicity substance to this activated gas.Wherein (usually in most cases to this antigenicity substance, can think to use the antigenicity substance of handling without activated gas) during for the comparing of this antibody in conjunction with activity, this antigenicity substance is active for the combination of this antibody, can adopt the measured value of measuring in advance, perhaps the measured value that also can adopt each enforcement evaluation method of the present invention to measure.Consider from the accuracy aspect of evaluation result, preferably adopt each measured value when implementing evaluation method of the present invention, but for easy and obtain evaluation result, the preferential measured value of measuring in advance of using rapidly.
Wherein, obtain the step of this processed antigen material, preferably include to make and be suspended in the step that aerial antigenicity substance and this activated gas react.
Like this by making the activated gas and the skyborne antigenicity substance reaction that suspends, antigenicity substance and activated gas are reacted under uniform state, have by regulating the suspension time of antigenicity substance, just have and to be easy to regulate the antigenicity substance and the advantage in activated gas reaction time.And, can stir or make it to flow to the atmosphere gas that contains activated gas for antigenicity substance is suspended in the air, just can make antigenicity substance fly up, and be suspended in aerial; Or antigenicity substance is fallen at a certain distance and it is suspended in the air.
Carry out the step of this reaction, be preferably included in the container step of scattering this antigenicity substance, make step that the solution that contains this distributions antigenicity substance suspends and the step of this activated gas of importing in this container in this container.
Like this,, can prevent the skimble-skamble diffusion of antigenicity substance, have the advantage that easily container endoantigen material concentration is kept within the specific limits by in container, scattering the solution that contains antigenicity substance.Wherein, container can be preferably airtight is container, but also can be that the semi-hermetic with part opening port is a container.
Be suspended in the container by the solution that contains this antigenicity substance that makes distribution like this, even contain the atmosphere gas of activated gas by stirring or make its flow impel antigenicity substance to fly up in, can prevent the skimble-skamble diffusion of antigenicity substance, have the advantage that easily container endoantigen material concentration is kept within the specific limits.
And by in container, importing the method for active gases like this, owing to can prevent the skimble-skamble diffusion of activated gas, therefore having can be in the container in antigenicity substance concentration keeps certain limit, the advantage that certain activated gas of concentration range and antigenicity substance are evenly reacted.
Wherein, owing to antigenicity substance is comprised among the solution, so when distribution contains the solution of antigenicity substance in container, preferably use sprayer to spray.This be since it can to spray particle diameter small and uniform solution can make the reaction of antigenicity substance and activated gas carry out more equably.
Activated gas of the present invention in addition makes the step of the aforementioned processed antigen material of acquisition in the method for evaluating performance of antigenicity substance inactivation, preferably include aforementioned antigenicity substance is applied vibration and/or impacts, make the aforementioned antigenicity substance skyborne step that suspends.This suspension step preferably includes the step that aforementioned antigenicity substance is arranged on the step on the sample bench with flexibility and sample bench is applied vibration and/or impacts.Wherein aforementioned sample bench with flexibility is to be selected from one or more of bed clothes, woollen blanket, cushion, pillow, mat, sponge, cloth, paper, styrenic foams.The step that aforementioned sample bench is applied vibration and/or impacts is in addition preferably beaten and/or is vibrated the said sample platform and method that sample bench is applied vibration and/or impacts.
In the step that obtains this processed antigen material, preferably include the step that makes this antigenicity substance and be selected from one or more gas reactions of the gas that contains positive ion, the gas that contains negative ion, the gas that contains free radical, ozone gas, nitric acid gas.Wherein, the step that especially preferably obtains this processed antigen material is the step that antigenicity substance and the gas that contains the both positive and negative ion are reacted.
Contain the gas of both positive and negative ion about this, as described later, by the inventor clear and definite first it have the function that makes the antigenicity substance inactivation that derives from Chinese fir pollen, and expect that it has the function that makes other antigenicity substance inactivation.In addition, for ozone gas, nitric acid gas, contain the gas of free radical, because they also are gaseous materials, so can estimate its deactivation by using the evaluation method in this instructions to antigenicity substance.
This determination step preferably includes to make at the antibody of this antigenicity substance and this processed antigen material by ELISA method and/or ELISA inhibition method and reacts, and measures the step of this processed antigen material for this antibody binding activity.
Like this, suppress method by ELISA method and/or ELISA, it is active for the combination of antibody accurately and easily to measure the processed antigen material.
For example as mentioned above by enzyme linked immunological absorption inhibition method, the antigenicity substance of measuring activated gas processing show 50% suppress concentration the time, can compare 50% inhibition concentration of this 50% inhibition concentration with the antigenicity substance of handling without activated gas.At this moment, if when described 50% inhibition concentration reaches 5 times, its remaining activity is decided to be 20%, (promptly reacting inactivation rate is 80%).
This determination step, preferably include by to except that the people, the animal that has at the antibody produced cell of this antigenicity substance carries out intradermal reaction test and/or conjunctival reaction test, this antibody and this processed antigen material are reacted, measure the step of this processed antigen material with respect to this antibody binding activity.
Like this by to except that the people, the animal that has at the antibody produced cell of this antigenicity substance carries out intradermal reaction test and/or conjunctival reaction test, also have and under in vivo status condition more, to measure the advantage of processed antigen material with respect to antibody binding activity near the people.Wherein described in postorder embodiment, though the people is carried out intradermal reaction test and conjunctival reaction test, if but when using the live test that the mammals except that the people such as mouse, rat and rabbit carry out carrying out in human body, test much easier more than personnel selection enforcement.This point is a technical general knowledge in fields such as medical science, pharmacy, agronomy, biology, biological chemistry, molecular biology.
The generating apparatus of＜treated antigenicity substance 〉
Make the generating apparatus of processed antigen material of evaluation sample of the performance of antigenicity substance inactivation as activated gas of the present invention, include container, in this container, scatter the device of antigenicity substance and in this container, take place or import this activated gas device.The generating apparatus of processed antigen material of the present invention in addition also can possess container, encloses the device of antigenicity substance and the device of this activated gas takes place or imports in this container in this container.
By using such device, can be easy under uniform state, activated gas and antigenicity substance are reacted, generate high-quality, very be suitable as the processed antigen material that activated gas makes the performance evaluation sample of antigenicity substance inactivation.The further generating apparatus of preferred processed antigen material of the present invention possesses and makes the device of antigenicity substance at this container inner suspension.Can prevent the diffusion of activated gas and antigenicity substance by the existence of container, even so by the atmosphere gas that contains activated gas being stirred or it being flowed, antigenicity substance is flown up be suspended in the container, the concentration of antigenicity substance and activated gas also can keep within limits.
Wherein, preferably this container has the partially transparent material or all is transparent material.
Like this, owing to be that part or all is the container of transparent material, can be by the suspended state of visualization internal tank antigenicity substance, so have the advantage of easy adjusting antigenicity substance and activated gas reaction conditions.
Fig. 2 is that another example of expression generates as estimating device sketch map sample, the processed antigen material that activated gas of the present invention makes the performance of antigenicity substance inactivation.
Device shown in Figure 2 possesses the cylindrical pressure vessel 1027 that has as the semi-hermetic type of container.Also have as the sprayer 1024 and the inlet 1028 that scatter the antigenicity substance device.Have as making the semi-hermetic type cylindrical pressure vessel 1027 of antigenicity substance, because it has certain altitude, so portion will inevitably form the state that antigenicity substance suspends within it at container inner suspension device.And possess ion generating device 1021 is arranged, contain the device of the gas of positive ion 1022, negative ion 1023 simultaneously as in this container, importing as activated gas.
In the device shown in Figure 2, in addition also show the returnable 1025 and the atmosphere gas exhausr port 1026 that contains activated gas of the antigenicity substance of handling by activated gas.
Fig. 3 is that the another example of expression generates as estimating the device sketch map of processed antigen material of sample that activated gas of the present invention makes the performance of antigenicity substance inactivation.
Device shown in Figure 3 possesses the semi-hermetic type cylindrical pressure vessel 1037 that has as container.Also possesses the inlet 1038 that has as the device that scatters antigenicity substance.Also have as making the semi-hermetic type container 1037 of antigenicity substance, because it has certain altitude, so portion will inevitably form the state that antigenicity substance suspends within it at container inner suspension device.And possess ion generating device 1031 is arranged, contain the device of the gas of both positive and negative ion as in this container, importing as activated gas.
Fig. 4 is that the another example of expression generates the device sketch map of processed antigen material of evaluation sample that makes the performance of antigenicity substance inactivation as activated gas of the present invention.
Device shown in Figure 4 possesses the hermetic type cylindrical pressure vessel 1047 that has as container.Also possesses the opening cap 1048 that has as the device that antigenicity substance is scattered.Also have as making the hermetic type cylindrical vessel 1047 of antigenicity substance at the device of container inner suspension, because it has certain altitude, so by making its setting along its length, or it is overturn repeatedly, portion will inevitably form the state that antigenicity substance suspends within it.And possess ion generating device 1041 is arranged, contain the device of the gas of both positive and negative ion as in this container, importing as activated gas.
In the device shown in Figure 4, in addition antigenicity substance 1049 is shown also, applies electrode 1042, dielectric 1043, the ground-electrode 1044 of voltage.
Fig. 5 is that the another example of expression generates as estimating the device sketch map of processed antigen material of sample that activated gas of the present invention makes the performance of antigenicity substance inactivation.
Device shown in Figure 5 possesses the hermetic type cylindrical pressure vessel 1057 that has as container.Also possesses the opening cap 1058 that has as the device that antigenicity substance 1053 is scattered.Also have and make antigenicity substance 1053 be suspended in the fan 1059 of device in the container.And possess ion generating device 1051 is arranged, contain the device of the gas of both positive and negative ion 1052 as activated gas as in this container, importing.
Fig. 6 is that the another example of expression generates the device sketch map of processed antigen material of evaluation sample that makes the performance of antigenicity substance inactivation as activated gas of the present invention.
Device shown in Figure 6 possesses the hermetic type cylindrical pressure vessel 1067 that has as container.Also possesses the opening cap 1068 that has as the device that antigenicity substance 1063 is scattered.Also have conduct and make the fan 1069 of antigenicity substance 1063 at container inner suspension device, and the filtrator 1065 that can only can not see through antigenicity substance through activated gas.And possess ion generating device 1061 is arranged, contain the device of the gas of both positive and negative ion 1062 as activated gas as in this container, importing.
＜ion generating device 〉
As estimating employed ion generating device in the generating apparatus of processed antigen material of sample of performance that activated gas of the present invention makes the antigenicity substance inactivation, be the element that positive ion and negative ion can take place, preferably can directly make the element of the allergic effect reaction inactivation of antigenicity substance by surge as described later.
Installation position for this ion generating device is not particularly limited, but is preferably mounted at usually on the ventiduct of the device that makes the antigenicity substance inactivation.This is because can be disappeared at short notice by the both positive and negative ion that ion generating device took place, so will guarantee to make these both positive and negative ions to be diffused in the air effectively during design.Ion generating device number is set, can be one, also can be two or more.
As this ion generating device, can use the known ion generating device that the both positive and negative ion takes place discharge mechanism that passes through.Particularly the ion generating device of Xuan Zeing will make positive ion and negative ion are acted in the atmosphere gas of antigenicity substance that the concentration of both positive and negative ion reaches 100,000/cm respectively when carrying negative ions in air 3Or more than it.And in this manual, the meaning of so-called ion concentration is meant small ion concentration, as the assay method of this small ion concentration, can adopt to make critical mobility reach 1cm 3/ V second, the value of measuring by air ion counter (ダ Application section length of schooling air ion counter (article number is 83-1001B)).
Wherein said discharge mechanism, it is the structure that has with electrode clamping insulator, apply the high voltage of interchange in a side, make the opposite side electrode grounding simultaneously, by applying high-tension electricity, form plasma discharge near ground-electrode the air layer, airborne hydrone and oxygen molecule are carried out ionization or disassociation and generate the mechanism of both positive and negative ion.In this discharge mechanism, it is tabular or netted making the electrode shape that applies voltage side, and when to make the ground connection lateral electrode be netted, if apply high voltage, then electric field was concentrated in the net end face area of ground connection lateral electrode, causes surface-discharge, forms the plasma zone.If air flows into these gas ions zones, just can generate the both positive and negative ion.
As element with this discharge mechanism, can enumerate as surface discharge element, corona discharge cells, plasma discharge element or the like, but also be not limited to this several elements.Aspect the electrode shape and material of arresting element, be not subjected to the qualification of above-mentioned shape and material in addition yet, can from the element of all shapes, material, select.
Fig. 7 is a structure example sketch map representing ion generating device used among the present invention.
As this ion generating device, more particularly preferably as shown in Figure 7, mesh electrode 7004 clamping dielectrics 7003 with plate electrode 7002 that applies voltage and ground connection, alternately plate electrode is applied positive pole and cathode voltage by high-voltage power supply 7001, electric field is concentrated at the net end face of mesh electrode, cause plasma discharge, form plasma zone 7005, generate the element of both positive and negative ionic structure.
In order to take place and to carry the needed impressed voltage of these both positive and negative ions, different and different with the structure of ion generating device, can be 2～10kV still as interelectrode peak to peak value (peak to peak) voltage range, preferred 3～7kV.
＜make the antigenicity substance inactivation by the gas that contains the both positive and negative ion 〉
The inventor uses as estimating the generating apparatus of processed antigen material of sample that activated gas of the present invention makes the performance of antigenicity substance inactivation, make the evaluation method of the performance of antigenicity substance inactivation by activated gas of the present invention, described in the embodiment of back, found that the gas that contains the both positive and negative ion has the function that makes the antigenicity substance inactivation.
But the present invention is not limited to negative ions, can also use as object with all gases or gas concentration.
Can think that the gas that contains the both positive and negative ion by use makes the mechanism of antigenicity substance inactivation, be not only aforesaid mechanism, but also comprise by the surge in the ion generating device mechanism that sex change or destruction cause inactivation is carried out at the antibody response position of antigenicity substance based on chemical reaction.
Just can think the antibody response position of antigenicity substance, when producing the both positive and negative ion owing to the effect that applies the plasma discharge self that voltage causes, also can produce sex change or be damaged, under this surge, the binding ability of antigenicity substance and antibody also can be lost, thereby makes the antigenicity substance inactivation.
Like this, make the evaluation method of the performance of antigenicity substance inactivation by activated gas of the present invention, be to carry out sex change or destruction by surge and/or chemical reaction to the antibody response position of antigenicity substance, thereby make the method for antigenicity substance inactivation, particularly, can obtain making the result of the effective inactivation of antigenicity substance by the two synergy of surge and chemical reaction.
＜contain the output intent of the gas of both positive and negative ion 〉
The inventor makes the evaluation method of the performance of antigenicity substance inactivation by activated gas of the present invention, as described later, what kind of method the gas of having found to contain the both positive and negative ion in use should preferably use contain the method for both positive and negative ionized gas as output during as activated gas actually.
Employed both positive and negative ion among the present invention just mainly is that the electric discharge phenomena by ion generating device take place, and usually, by alternately applying generating positive and negative voltage, almost the both positive and negative ion can take place simultaneously, and be transported in the air.But the method for conveying both positive and negative ion of the present invention, be not limited thereto, can also only apply any one voltage in the generating positive and negative voltage at first within a certain period of time, only export any one ion in the negative ions, and then applying the opposite voltage of certain hour, output has and the ion of having exported the ion opposite charges.
Wherein, take place and export the necessary impressed voltage of this both positive and negative ion, different and different with electrode structure, but can be 2～10kV as interelectrode peak to peak value voltage range, preferred 3～7kV.
In addition, used positive ion and negative ion among the present invention is 20～90% in relative humidity preferably, and more preferably relative humidity takes place under 40～70% condition.The generation of both positive and negative ion as described later exists relevant with airborne hydrone.Just when relative humidity is lower than 20%, can not suitably carry out the cohesion that causes by the hydrone that with the ion is the center, cause combination again between the ion easily, so the ion lifetime that is taken place shortens.And when relative humidity surpasses 90%, because moisture is at the ion generating device surface sweating, the luminous efficiency of ion obviously reduces, and also excessively carrying out of the ion that is taken place because of cohesion, and surrounded by many hydrones, so make the weight of ion increase too much, might just produce sedimentation at a distance not under the state of carrying.Therefore, it all is unfavorable under extremely low in humidity as mentioned above or the state that humidity is high ion taking place.
As the output intent of both positive and negative ion of the present invention, the method for the employing electric discharge phenomena of narrating is previously not only arranged, can also adopt the method that can send ultraviolet ray and electron beam apparatus etc.
The evaluation of＜both positive and negative ion 〉
Among the present invention, the gas that contains the both positive and negative ion when use is during as activated gas, and positive ion and negative ion can be that raw material takes place with oxygen molecule and/or the hydrone that is present in the arresting element surface.If according to this method for generation, owing to do not need special raw material, and favourable aspect cost.Moreover, self does not have harmfulness raw material, can not produce other harmful ion and material, so be preferred.
Wherein, the composition of the both positive and negative ion that the electric discharge phenomena by above-mentioned ion generating device take place as positive ion, mainly is by plasma discharge, makes airborne hydrone ionization, generates hydrogen ion H +, this hydrogen ion H +Under the effect of solvation energy, condense with airborne hydrone, form H 3O +(H 2O) n(n be 0 or natural number).If wherein change a kind of method for expressing, as the H of positive ion statement 3O +(H 2O) n(n be 0 or natural number) then can be expressed as H +(H 2O) n(n is a natural number) also represents identical ion.
Fig. 8 A and 8B are that expression is by the positive ion of ion generating device generation and the mass spectrum of negative ion.
From Fig. 8 A, can observed smallest peaks be positioned at the position of molecular weight 19, the peak of back appears at respect to this molecular weight 19 and adds the position that is equivalent to molecular weight water 18 in turn, shows that thus hydrone condenses.Just this result shows at molecular weight to be 1 hydrogen ion H +On, molecular weight is hydrone and integral body of its formation of 18, carries out hydration.As negative ion, by plasma discharge, airborne oxygen molecule or hydrone carry out ionization on the other hand, generate oxonium ion O 2 -, this oxonium ion O 2 -Under the effect of solvation energy, form O with airborne hydrone cohesion 2 -(H 2O) m(m be 0 or natural number).From Fig. 2 (b), can observed smallest peaks be positioned at the position of molecular weight 32, the peak of back appears at respect to this molecular weight 32 and adds the position that is equivalent to molecular weight water 18 in turn, shows that thus hydrone condenses.Just this result shows at molecular weight to be 32 oxonium ion O 2 -On, molecular weight is hydrone and integral body of its formation of 18, carries out hydration.
And these both positive and negative ions that are transported to the space have surrounded and have been suspended in airborne antigenicity substance, can infer on the antigenicity substance surface, and the both positive and negative ion generates the hydrogen peroxide H of reactive specy by following chemical reaction (1)～(2) 2O 2, Liquor Hydrogen Peroxide (hydrogendioxide) HO 2Or hydroxyl radical free radical OH.
H 3O ++O 2 -→·OH+H 2O 2 ...(1)
H 3O ++O 2 -→HO 2+H 2O ...(2)
Can understand the hydrogen peroxide H that such both positive and negative ionization generates like this 2O 2, Liquor Hydrogen Peroxide HO 2Or hydroxyl radical free radical OH, sex change or destruction (decomposition) are carried out in the antibody position of antigenicity substance, make the binding ability forfeiture of antigenicity substance and antibody can make airborne antigenicity substance inactivation effectively.
In the above description, respectively with H as positive ion 3O +(H 2O) n(n be 0 or natural number) is as the O of negative ion 2 -(H 2O) m(m be 0 or natural number) is for narrating at the center, but the negative ions among the present invention is not subjected to their qualification.Based on above-mentioned two kinds of negative ions,, can also enumerate N as positive ion 2 +, O 2 +Deng; Can enumerate NO as negative ion 2 -, CO 2 -Deng.Also can expect to obtain identical effect even contain these ions.
Embodiment
Be described in more detail situation of the present invention below by embodiment, but the present invention is not subjected to the qualification of these embodiment.
＜Chinese fir pollen 〉
Gather Chinese fir pollen from branch the Japanese Chinese fir (Cryptomeria japonica) of Hiroshima Xian Fengding growth.Adopt the suction cleaner that screen cloth is housed in the gatherer process, collection pollen then sieves.Collecting the back adopts-30 ℃ refrigerator to preserve.
＜Chinese fir antigenicity substance 〉
Chinese fir pollen 80g in 20mM PBS (pH7.4) 3.2L, under 4 ℃, is stirred after 4 hours, carry out 30 minutes centrifuging with the rotating speed of 6000rpm.After the centrifuging, in supernatant, add ammonium sulfate, and make its ultimate density reach 80% saturated,, carry out 30 minutes centrifuging with the rotating speed of 6000rpm.After the centrifuging, carry out 6 times 6 hours dialysis treatment repeatedly, with the rotating speed of 10000rpm, carry out 30 minutes centrifuging again.After the centrifuging, the gained supernatant is carried out freeze drying, as the Chinese fir antigenicity substance.In this instructions, Chinese fir antigenicity substance note is made CJP.
＜the protein content that carries out with the Folin-Lowry method is measured 〉
[composition of reagent]
A liquid: with the phenol reagent of 1N as acid solution
B liquid: 2%, Na 2CO 3
0.1N NaOH
C liquid: 0.5%, CuSO 45H 2O
1% sodium citrate
D liquid B: C=50: the mixed liquor of 1 (v/v)
[assay method]
Sample 0.2ml and D liquid 1ml are mixed, placed 10 minutes.Then add A liquid 0.1ml and place after 30 minutes, measure its absorptance at the 750nm place.Also make standard series, and prepare calibration curve, the protein content of sample is carried out quantitatively as BSA conversion amount with same sequence with BSA.
The distribution of＜Chinese fir antigenicity substance, recovery 〉
The Chinese fir antigenicity substance (protein concentration 200ng/ml) that extracts from Chinese fir pollen under the irradiation of negative ions, is scattered with sprayer.In the bottom of scattering container the recovery ware is set, can only reclaims and do not contact and through the antigen of ion processing with the container wall.With 1.5 hours distribution 8ml solution (containing the Chinese fir antigenicity substance).
＜embodiment 1 〉
Present embodiment is the antigenicity substance that adopts Chinese fir pollen, confirms to cause the situation that the allergic effect reaction of antigenicity substance reduces through the both positive and negative ionization.
Wherein Fig. 2 enumerates an example explanation as estimating the sketch map of processed antigen material generating apparatus of sample that activated gas of the present invention makes the performance of antigenicity substance inactivation.Fig. 8 A, 8B are that expression is by the positive ion of the ion generating device generation that is provided with in the device shown in Figure 2 and the mass spectrum of negative ion.
At first in device shown in Figure 2, as ion generating device 1021, use be long 37mm, the tabular surface discharge element of wide 15mm.And partly cause surface-discharge by between electrode, alternately applying the voltage of positive and negative, making at surface electrode,, generate and export positive ion 1022 and negative ion 1023 simultaneously by plasma discharging district under atmospheric pressure.Alive interelectrode peak to peak value voltage range is 3.3kV～3.7kV.In this voltage range, can not produce the ozone of harmful level.At internal diameter is that 150mm, length are in the cylindrical vessel 1027 of semi-hermetic type of acrylate system of 370mm, install and fix 4 such ion generating devices, side at this container is provided with the inlet 1028 that scatters antigenic substance solution, and the returnable 1025 of antigenicity substance liquid is set at the opposite side of this container.
When using the antigenic substance extract from Chinese fir pollen as antigenic substance, Chinese fir pollen is to gather from the branch the Japanese Chinese fir (Cryptomeria japonica) of Hiroshima Xian Fengding growth.Adopt the suction cleaner that screen cloth is housed in the gatherer process, collection pollen then sieves.Collecting the back adopts-30 ℃ refrigerator to preserve.The method of from Chinese fir pollen, extracting antigenic substance be Chinese fir pollen 80g in 20mM PBS (pH7.4) 3.2L, under 4 ℃, stirred 4 hours, carry out 30 minutes centrifuging then with the speed of 6000rpm.After the centrifuging, in supernatant, add ammonium sulfate, and make its ultimate density reach 80% saturated,, carry out 30 minutes centrifuging with the rotating speed of 6000rpm.After the centrifuging, carry out 6 times 6 hours dialysis treatment repeatedly, with the rotating speed of 10000rpm, carry out 30 minutes centrifuging again.After the centrifuging, the gained supernatant is carried out freeze drying, as antigenic substance liquid.
Antigenic substance liquid 8ml for test is put in the sprayer 1024, and the inlet of using with the distribution antigenicity substance of device shown in Figure 2 1028 links.The returnable 1025 of the antigenic substance liquid of same device is arranged on the bottom of semi-hermetic type cylindrical vessel 1027.Sprayer and air compressor link, and scatter for the antigenicity substance of testing from inlet 1028 by pressurized air (flow 5L/ minute).Dispersion volume is 8.0ml (the distribution time is 90 minutes).Capture the antigenicity substance that is deposited in semi-hermetic type cylindrical vessel bottom in 90 minutes by returnable.The antigenicity substance of being sprayed falls naturally through 90 seconds in air, with airborne positive ion 1022 and negative ion 1023 effects.
Measure by the ELISA method pair and the reactivity of the SERUM IgE antibody of gathering from the pollinosis patient.Wherein the concentration of both positive and negative ion is the inlet of using from the distribution antigenicity substance liquid of the semi-hermetic type cylindrical vessel 1027 that is provided with ion generating device 1021 as described above 1028, by the flow input air of air compressor with 5L/ minute, the air ion counter (article number is 83-1001B) that the manufacturing of ダ Application science is set in antigenicity substance liquids recovery container 1025 is measured the total concentration of the both positive and negative ion in this space.The temperature of space atmosphere is 25 ℃, and relative humidity is 60%RH.Can infer that shown in Fig. 8 A, 8B, the positive ion that is output is H 3O +(H 2O) n(n be 0 or any natural number), negative ion is O 2-(H 2O) m(m be 0 or any natural number), this both positive and negative ion is by aforementioned chemical reaction (1) and (2) generation hydrogen peroxide H 2O 2, Liquor Hydrogen Peroxide HO 2Or hydroxyl radical free radical OH.
State when ion generating device 1021 is not worked is decided to be the state of being untreated, to this element voltage that to apply interelectrode peak to peak value voltage respectively be 3.3kV～3.7kV, output both positive and negative ion, research makes the concentration of the both positive and negative ion of semi-hermetic type cylindrical vessel 1027 inside reach the both positive and negative ion and is respectively 100,000/cm 3The time, the reactive situation about reducing of the allergic effect of antigenicity substance and IgE antibody.The result is shown in Fig. 9 A, 9B and Figure 10 A, 10B.
Fig. 9 A, Fig. 9 B are expressions with containing the gas of both positive and negative ion, when the Chinese fir antigenicity substance is handled and when being untreated, and the chart of the allergic effect reaction relation of antigenicity substance and pollinosis patient's 19～40 SERUM IgE antibody.
Figure 10 A, Figure 10 B are expressions with containing the gas of both positive and negative ion, when the Chinese fir antigenicity substance is handled and when being untreated, and the chart of the allergic effect reaction relation of antigenicity substance and pollinosis patient's 41～60 SERUM IgE antibody.
Shown in Fig. 9 A, 9B and Figure 10 A, 10B, can confirm (just not take place under the state of negative ions) when ion generating device is not worked and the concentration of both positive and negative ion reaches 100,000/cm respectively 3The time, the reactivity (associativity) of pollen patient's SERUM IgE antibody has 33 people's SERUM IgE antibody response to present significant reduction in pollen patient 42 people.
After scattering with sprayer, when not making being untreated of ion generating device work, and to this element voltage that to apply interelectrode peak to peak value voltage respectively be 3.3kV～3.7kV, and the output negative ions, make the negative ion concentrations in the semi-hermetic type cylindrical vessel 1027 reach 100,000/cm respectively 3The time, the reactivity reduction situation of Cry j 1 and Cry j 2 monoclonal antibodies and SERUM IgE antibody is studied.The result as shown in figure 11.
Figure 11 is when representing to use the gas that contains negative ions that the Chinese fir antigenicity substance is handled and when being untreated, and the chart of antibody response sexual intercourse falls in Cry j 1 and Cry j 2 and its Dan Ke.
Can confirm when ion generating device is not worked, (state of negative ions just not to take place) and the both positive and negative ion reaches 100,000/cm respectively 3The time, the reactivity (associativity) of pollen patient's SERUM IgE antibody, when carrying out ion processing, the SERUM IgE antibody response that antibody falls in Cry j 1 and Cry j 2 Dan Ke presents significant reduction.
For to carrying out quantitative evaluation, adopt ELISA inhibition method to test through ion processing with without the Chinese fir antigenicity substance of ion processing and the reactivity difference of pollinosis patients serum IgE.
Specifically be that the Chinese fir antigenicity substance that reclaim spraying back is put in the centrifugal separator (Centriprep YM-10), carry out centrifugal concentrating with the rotating speed of 2500rpm.Again this concentrate is put in the centrifugal separator (ULTRA FLEE-MC), carries out centrifugal concentrating with the 7000rpm rotating speed.Ion processing Chinese fir antigenicity substance and undressed Chinese fir antigenicity substance to concentrating carry out 85 times of dilutions repeatedly from protein concentration 11 μ g/ml.Each antigenicity substance 50 μ l that will dilute respectively and the patients serum IgE50 μ l of 10 times of dilutions mix, 4 ℃ of one nights of following preincubate.
Apply the Chinese fir antigenicity substance (also can not scattering) that is diluted to 1 μ g/ml with bicarbonate buffer (Bicarbonatebuffer) at ELISA on the 96-orifice plate, every hole adds 50 μ l, static placement 2 hours.With lavation buffer solution (Washing buffer) plate is carried out 3 washings, apply sealing damping fluid (Blocking buffer) 300 μ l then, 4 ℃ of one nights of down static placement.After plate carried out 3 washings, add sample 50 μ l through preincubate, static placement 4 hours to every hole respectively.
After plate carried out 3 washings, apply through (the biotin labeled anti human IgE that 3% skimmed milk+1%BSA)/PBST dilution is 1000 times, every hole 50 μ l, static placement 2.5 hours.After plate carried out 3 washings, apply through (the streptavidin 50 μ l of the alkali phosphatase enzyme mark that 3% skimmed milk+1%BSA)/PBST dilution is 1000 times, at room temperature static placement 1.5 hours.
To plate carry out 4 times the washing after, apply the AttophosTM substrate buffer solution, every hole 50 μ l are placed to color development under the shading state.Measure fluorescence intensity by CytoTMF luorII.When not making being untreated of ion generating device work and the voltage that to apply as interelectrode peak to peak value voltage in this element respectively be 3.3kV～3.7kV, output both positive and negative ion makes the concentration of both positive and negative ion in the semi-hermetic type cylindrical vessel 1027 reach 100,000/cm respectively 3The time, the reactivity (associativity) of pollen patient's SERUM IgE antibody is studied.Its result as shown in figure 12.
Figure 12 is illustrated in when using the gas processing Chinese fir antigenicity substance that contains the both positive and negative ion and when being untreated, and by enzyme linked immunological absorption inhibition method, measures antigenicity substance and pollinosis patient's the reactive graph of a relation of SERUM IgE antibody allergic effect.
(state of negative ions does not just take place) when ion generating device is not worked, the necessary Chinese fir antigenicity substance amount of 50% inhibition is 2.53 * 10 3Pg; In contrast, be respectively 100,000/cm when the both positive and negative ion concentration 3The time, the necessary Chinese fir antigenicity substance amount of 50% inhibition is 1.34 * 10 4Pg can confirm that its reaction inactivation rate is 81%.
By the tuberculin syringe, respectively with 0.9%NaCl to through ion processing with to be diluted to protein concentration without the Chinese fir pollen-antigen material of ion processing be the inboard intracutaneous of forearm that the dilution 0.02ml of 0.5 μ g/ml is expelled to Chinese fir pollinosis patient, measure the erythema that occurs, the major diameter and the minor axis of tumor after about 15 minutes, by its mean diameter evaluation response.The result is as shown in table 1.
Table 1
Figure C200480010422D00241
Being designated as of erythema＜10mm-, erythema be being designated as of 10-20mm ±, erythema be being designated as of 20-30mm, tumor＜10mm+, is erythema that 30-40mm, tumor are being designated as of 10-14mm ++, erythema 40mm, tumor 15mm and tumor being designated as of pseudopodium appears +++.As shown in table 1, under the idle situation that is untreated of ion generating device (just not producing the state of negative ions) and in the both positive and negative ion concentration, reach 100,000/cm respectively 3Situation under, can confirm that pollinosis patient's intradermal reaction presents significant reduction.
Again by pipettor being that the dilution 5 μ l of 0.5 μ g/ml splash in Chinese fir pollen patient's the eye to be diluted to protein concentration through ion processing with without the Chinese fir antigenicity substance of ion processing with 0.9%NaCl, after about 15 minutes, observation is as the semilunar fold of conjunctival reaction, the hyperemia of eyelid skin and bulbar conjunctiva, the degree of itching, situation such as shed tears.During judgement, be decided to be can't see congested situation fully-, the situation with gargalesthesia that can see hyperemia a little is decided to be ±, the top of bulbar conjunctiva or bottom one side occur congested being decided to be+, congested being decided to be all appearred in the upper and lower of bulbar conjunctiva ++, occurring congested being decided to be in the whole bulbar conjunctiva +++, being decided to be of edema appearred in eyelid ++ ++.As shown in table 1 observations.
As shown in table 1, reach 100,000/cm respectively in the idle situation that is untreated of ion generating device (just not producing under the state of negative ions) with in the both positive and negative ion concentration 3Situation under, can confirm that pollinosis patient's conjunctival reaction presents significant reduction.
＜embodiment 2 〉
The SERUM IgE of using patient 19 in above-mentioned enzyme linked immunological absorption (ELISA) method is as antibody, make antigenicity substance (Chinese fir antigenicity substance) concentration (as protein concentration) be respectively 4 kinds of concentration of 100ng/ml, 200ng/ml, 400ng/ml, 800ng/ml, (just device adopts above-mentioned device shown in Figure 3 according to operation same as described above; When carrying out ion processing, negative ion concentrations is respectively 100,000/cm 3), obtain untreated Chinese fir antigenicity substance respectively and through the fluorescence intensity of the Chinese fir antigenicity substance of ion processing by enzyme linked immunosorbent assay.And, obtain the reaction inactivation rate of allergic effect reaction according to following calculating formula (3) by this fluorescence intensity.As shown in table 2 the result.
Table 2
Antigenicity substance concentration (ng/ml) 100 200 400 800
Reaction inactivation rate (%) 94 83 78 66
Reaction inactivation rate %=(1-C/D) * 100 ... (3)
C: through the fluorescence intensity of the Chinese fir antigenicity substance of ion processing
D: the fluorescence intensity of undressed Chinese fir antigenicity substance
Then, be that the situation of 200ng/ml is selected as benchmark with the concentration of above-mentioned antigenicity substance, exist under the prerequisite of following relation between ion concentration and the antigenicity substance concentration, obtain different negative ion concentrations and the relation of reacting inactivation rate.Just can think certain if react inactivation rate, between ion concentration and antigenicity substance concentration, just there is certain relation so, it is certain for example to establish ion concentration, antigenicity substance concentration is reduced to the state of half, antigenicity substance concentration is certain with establishing, ion concentration is increased under 2 times the state, can obtains identical reaction inactivation rate.Therefore, be that negative ions is respectively 100,000/cm with above-mentioned experiment intermediate ion concentration 3And above-mentioned antigenicity substance concentration is that two points of 200ng/ml are benchmark, and the different negative ion concentrations and the relation of reaction inactivation rate are shown in Figure 13.Just among Figure 13, negative ion concentrations is 2.5 ten thousand/cm 3, 50,000/cm 3, 100,000/cm 3, 200,000/cm 3Data, the data (horizontal ordinate of Figure 13 is represented the variable concentrations of negative ions) when corresponding respectively to antigenicity substance concentration 800ng/ml in the above-mentioned enzyme linked immunosorbent assay, 400ng/ml, 200ng/ml, 100ng/ml.
Shown in Figure 13 is clear and definite,, then reacts inactivation rate and also improve, if particularly negative ion concentrations is respectively 50,000/cm if negative ion concentrations increases 3, then react inactivation rate and can reach about 78%, can obtain the effect of stable antigenicity substance inactivation.If negative ion concentrations is respectively 100,000/cm in addition 3, then react inactivation rate and can reach 83%, if further make negative ion concentrations be respectively 200,000/cm 3, then react inactivation rate and can reach 94%, can expect effectively to suppress allergic diseases such as pollinosis and tick allergy.
In embodiment 1 and embodiment 2, used the gas that contains the both positive and negative ion as activated gas; Used the antigenicity substance that derives from Chinese fir pollen as antigenicity substance, if but the activated gas of the application of the invention makes the evaluation method of the performance of antigenicity substance inactivation, for the activated gas of other kind and the antigenicity substance of other kind, can accurately and easily the performance of antigenicity substance inactivation be estimated too to activated gas.
In addition in embodiment 1 and embodiment 2, use as shown in Figure 2 conduct activated gas of the present invention to make the generating apparatus of processed antigen material of the performance evaluation sample of antigenicity substance inactivation, generated the processed antigen material, generate the processed antigen material, can accurately and easily the performance of antigenicity substance inactivation be estimated too even use activated gas as Fig. 3～device shown in Figure 6.
＜embodiment 3 〉
Present embodiment is to use the antigenicity substance of tick dust, confirms to make by the effect of both positive and negative ion the situation of antigenicity substance inactivation.Followingly describe with reference to Figure 14 and Figure 15.
Figure 14 is a device sketch map of implementing to make by positive ion and negative ion effect the antigenicity substance method for deactivating.To be expression amount to the figure that 18 SERUM IgE reactivity is estimated by enzyme linked immunological absorption (ELISA) method to tick antigenicity substance (abbreviating Derf as) and patient a～r to Figure 15.The device of Figure 14, the same with the device of Fig. 2, have ion generating device shown in Figure 7, the positive ion by the output of this element and the mass spectrum of negative ion are shown in Fig. 8 A, 8B.
＜enforcement makes the device of antigenicity substance method for deactivating 〉
At first in the present embodiment used device shown in Figure 14 and device shown in Figure 2 be the same (therefore, the part that indicates identical reference marks among Fig. 2 and Figure 14 is represented same section or considerable part), just possess on the device this point that reduces ozone concentration different.In the device just shown in Figure 14, the exhausr port 1026 of a side and sprayer 1024 link by filtrator 1029.This filtrator 1029 includes activated charcoal and molecular sieve, has the effect of removing the ozone that is taken place in cylinder type closed container 1027.Therefore the ozone concentration in this cylinder type closed container 1027 can maintain 0.025ppm or below it.
In device shown in Figure 14, the same with the device of Fig. 2, antigenicity substance 1038 is sprayed into by inlet 1028, drops to naturally then on the returnable 1025, is exposed in the meantime in the both positive and negative ion, and is subjected to its effect.
＜tick dust and antigenicity substance 〉
As antigenicity substance, use by the antigenicity substance that extracts in the tick dust.The tick dust is an object with the tick dust that generally is present in the family, by the suction cleaner of screen cloth is installed, collects from cushion and flannelette blanket.
In order from the tick dust, to extract antigenicity substance, tick dust 0.1g is placed the phosphate buffer solution of 20mM, and (PBS is pH7.4) among the 15ml, under 4 ℃ temperature conditions, stir after 16 hours, make its pass through film filter (0.2 μ m) the material that obtains as the tick antigenicity substance.Also contain antigenicity substance Derf1 and Derf2 in this tick antigenicity substance.
＜the quantification of protein that undertaken by the Folin-Lowry method 〉
Solution 0.2ml that contains the tick antigenicity substance and following D liquid 1ml are mixed, placed 10 minutes.Then add following A liquid 0.1ml, place after 30 minutes, measure its absorptance at the 750nm place.Use bovine serum albumin matter (BSA) preparation standard series again, prepare calibration curve,, the amount of the protein of tick antigenicity substance is carried out quantitatively as BSA conversion amount according to same sequence.Its protein concentration is 94.1ng/ml as a result.Wherein used all ingredients is as follows:
(reagent)
A liquid: with the phenol reagent of 1N as acid solution.
B liquid: 2%Na 2CO 3The NaOH of+0.1N
C liquid: 0.5%CuSO 45H 2The O+1% sodium citrate
D liquid: B liquid: C liquid=50: 1 (v/v)
The spraying of＜antigenicity substance and recovery 〉
The solution that antigenicity substance is the tick antigenicity substance (protein concentration the is 200ng/ml) 8ml that contains that obtains is like this put in the sprayer 1024, be attached on the inlet 1028 that the antigenicity substance solution spray that installs shown in Figure 14 uses.In order to reclaim the solution that contains the antigenicity substance of spraying to some extent, returnable 1025 is set on the other hand in the bottom of cylinder type closed container 1027.
To spray medicine atomizer and air compressor and link, spray from 1028 pairs of antigenicity substances 1038 of inlet by pressurized air (flow is 5L/ minute).Spray amount is 8.0ml (spray time 90 minutes).After 90 minutes, reclaim the antigenicity substance that is deposited in cylinder type closed container 1027 bottoms by returnable 1025.The antigenicity substance 1038 of spraying falls naturally by cylindrical pressure vessel 1027 and needed for 90 seconds approximately.
Spraying and reclaim this antigenicity substance 1038 is when making ion generating device 1021 work (when carrying out ion processing) and when it is not worked (when not handling), carries out twice.
Make ion generating device 1021 work, when making positive ion and negative ion act on antigenicity substance, the both positive and negative ion concentration of (just in the cylinder type airtight container 1027) is following obtaining in its atmosphere: from the inlet of being used by the antigenicity substance solution spray of the cylinder type closed container 1027 that ion generating device 1021 is set 1028, pass through air compressor, air is flow through with 5L/ minute flow, and the air ion counter (article number is ITC-201A) that the electric manufacturing of ア Application デ ス is set on the returnable 1025 of antigenicity substance solution is measured the concentration of both positive and negative ion.When the result applied interelectrode peak to peak value voltage respectively and is the voltage of 3.3kV～3.7kV this ion generating device 1021, the both positive and negative ion concentration in the cylinder type closed container 1027 in the atmosphere reached 100,000/cm respectively 3Other condition of space atmosphere is that temperature is 25 ℃, and relative humidity is 60%RH.Can infer that shown in Fig. 8 A, 8B, the positive ion that is output is H3O +(H 2O) n, (n be 0 or natural number), negative ion is O 2 -(H 2O) m(m be 0 or natural number), this both positive and negative ion generates hydrogen peroxide H by aforementioned chemical reaction (1)～(2) 2O 2, Liquor Hydrogen Peroxide HO 2And hydroxyl radical free radical OH.
＜carry out reactive evaluation by enzyme linked immunological absorption (ELISA) method 〉
The reactivity of the SERUM IgE antibody of then collecting the tick antigenicity substance that obtains like this and gathering from tick autopath a-r by enzyme-linked immunosorbent assay.For antigenicity substance,, estimate its reactivity by to comparing through the treated substance (ion processing tick antigenicity substance) of aforesaid negative ions effect and the material that is untreated (undressed tick antigenicity substance).
Specific practice is to apply ion processing tick antigenicity substance and the untreated tick antigenicity substance that is diluted to 0.1 μ g/m1 with sodium bicarbonate buffer liquid (Bicarbonate buffer), every hole 50 μ l on enzyme linked immunological adsorbs with 96 orifice plates (ELISA 96-orifice plate).Simultaneously with sodium bicarbonate buffer liquid since 200 μ g/ml, (human IgE standard) carries out 52 times of dilutions repeatedly to people IgE standard, and dilution is added to respectively in the hole, every hole 50 μ l, at room temperature static placement 2 hours.With buffer solution (Washing Buffer) plate is carried out 3 washings with washing, apply damping fluid (Blocking buffer) the 300 μ l of sealing usefulness then, descend one night of static placement at 4 ℃.
After one night of static placement, plate is carried out 3 washings, add through with (3% skimmed milk+1%BSA)/PBST is to 20 times of tick autopath's serum dilutions, and carries out the material 50 μ l/ holes of hatching in 1 hour, static placement 4 hours to every hole.After plate carried out 3 washings, apply through (the biotin labeling anti human IgE that 3% skimmed milk+1%BSA)/PBST dilution is 1000 times, every hole 50 μ l, static placement 2 hours.
After this static placement, plate is carried out 4 washings, apply then through (the alkali phosphatase enzyme mark streptavidin 50 μ l that 3% skimmed milk+1%BSA)/PBST dilution is 1000 times, at room temperature static placement 1 hour.To plate carry out 5 times the washing after, apply Attophos (registered trademark) substrate buffer solution, every hole 50 μ l are placed to color development under the shading state.(Cyto (registered trademark) FluorII) measures its fluorescence intensity by spectrophotometer.The result as shown in figure 15.
As shown in figure 15, can confirm that (state that is untreated of both positive and negative ion does not just take place) and both positive and negative ion concentration do not reach 100,000/cm respectively when making ion generating device 1021 work 3The time, the reactivity (associativity) of tick autopath SERUM IgE antibody and tick antigenicity substance is in all 18 people of tick autopath a～r, significantly reduce (fluorescence intensity is low more, and the expression reactivity is low more) through the antigen of aforementioned processing and above-mentioned patient's SERUM IgE antibody reactive.Wherein used all ingredients is as follows:
(reagent)
The NaHCO of sodium bicarbonate buffer solution: 100mM 3(pH9.2～9.5),
Phosphate buffer solution (PBS): use distilled water the Na of the NaCl of 4g, 0.1g 2HPO 412H 2The KCl of O, 1.45g, the KH of 1g 2PO 4It is fixed molten to 500ml,
PBST:PBS+0.5% Tween-20 (Tween-20),
Sealing buffer solution: PBS+3% skimmed milk+1%BSA,
Washing buffer solution: use distilled water the Na of 43g 2HPO 412H 2The NaH of O, 3.6g 2PO 4, 263g Tween-20 (Tween-20) constant volume of NaCl, 15ml to 3L.
＜reaction inactivation rate 〉
The SERUM IgE of using the patient a～r in above-mentioned enzyme linked immunological absorption (ELISA) method is as antibody, obtain untreated tick antigenicity substance respectively and through the fluorescence intensity of the tick antigenicity substance of ion processing by enzyme linked immunosorbent assay, and use this fluorescence intensity, obtain the reaction inactivation rate of allergic effect reaction according to following calculating formula (4).The result is shown in following table 3.
Table 3
Figure C200480010422D00311
Reaction inactivation rate %=(1-E/F) * 100 ... (4)
E: through the fluorescence intensity of the tick antigenicity substance of ion processing
F: the fluorescence intensity of undressed tick antigenicity substance
Table 3 shows that the average response inactivation rate of patient a～r is 57.8%, can expect to obtain effectively to suppress the effect of tick allergic disease.
＜embodiment 4 〉
In the present embodiment, directly use the tick dust, confirm inactivation situation by the tick dust (wherein contained antigenicity substance) of negative ions effect processing.Describe with reference to following Figure 11～13.It is identical with embodiment 3 wherein by the Folin-Lowry method albumen quality in the tick antigenicity substance contained in the tick dust to be carried out quantitative operation.
The diffusion of＜tick dust and recovery 〉
The diffusion of tick dust is carried out (wherein indicate among Figure 16 have with other figure identical reference marks represent same section or considerable part) with reclaim adopting as shown in figure 16 device.Just this device is provided with ion generating device 1021 by fan blower 1033 being installed and operating and form with the closed box 1030 of window 1034 at the air ejiction opening place of this fan blower 1033.
At first make ion generating device 1021 work, meanwhile make fan blower 1033 work.Its condition is that interelectrode peak to peak value voltage-regulation with this ion generating device 1021 is to 90V, so that the space average concentration of both positive and negative ion reaches 3000/cm respectively 3, and the air output of this fan blower 1033 transferred to 2m 3/ minute.
The space average concentration of the both positive and negative ion in this closed box 1030, be the concentration of measuring the both positive and negative ion of near the phase mutual edge distance 50cm in this closed box center or its 5 above points by the air ion counter (article number is ITC-201A) that ア Application デ ス electric corporation is made respectively, make the mean value of both positive and negative ion reach 3000/cm respectively 3Space atmosphere temperature in this closed box 1030 is 25 ℃ in addition, and relative humidity is 60%RH.Can infer that shown in Fig. 8 A, 8B, the positive ion that is output is H 3O +(H 2O) n(n be 0 or natural number), negative ion is O 2 -(H 2O) m(m be 0 or natural number), this both positive and negative ion generates hydrogen peroxide H by aforementioned chemical reaction (1)～(2) 2O 2, Liquor Hydrogen Peroxide HO 2With hydroxyl radical free radical OH.
The space average concentration of the so-called both positive and negative ion among the present invention, be meant the mean concentration in whole space with certain volume, for example can be determined at respectively near the indoor center of suitable air trapping by ion counter (for example ア Application デ ス electric corporation make air ion counter (article number is ITC-201A)), the concentration of the both positive and negative ion of phase mutual edge distance 50cm or its 5 above points is measured by the mean concentration of obtaining these 5 points then.
Ion generating device 1021 and this fan blower 1033 are temporarily quit work, the article 1032 that support tick dust (2g) are set in this closed box 1030 then, under condition same as described above, make ion generating device 1021 and fan blower 1033 work again.
Follow by action pane 1034, use diffusion instrument 1035 to beat article 1032, make tick dust 1031 diffusions (scattering or suspension).Wherein, as article 1032, can enumerate bed clothes, woollen blanket, flannelette blanket, paillasse, pillow, cushion, back cushion or the like.Adopted cushion in the present embodiment.As diffusion instrument 1035, can enumerate the bat that beats bed clothes, duster, broom or the like.Used the bat that beats bed clothes in the present embodiment.As dispersion operation, except beaing article 1032, the method that can also adopt shake or it is fallen.Be to adopt to beat bat that bed clothes use as diffusion instrument 1035 in the present embodiment, article 1032 cushions are firmly beaten that the time is 5 minutes, amounting to and beaing number of times is 20 times.
After then cushion being beaten end, make suction pump 1037 work that are arranged on these closed box 1030 tops, by reclaiming the dust in the filtrator 1036 collected at suction seal boxs 1030, the collected at suction time is 30 minutes.
After 30 minutes, suction pump 1037 is quit work, the bat with diffusion instrument 1035 beat 5 minutes article 1032 cushions once more, beat altogether 20 times.And then make suction pump 1037 work, by reclaiming the dust in the filtrator 1036 collected at suction closed boxs 1030, suction time is 30 minutes.
To according to aforesaid operations,, be 0.7mg with reclaiming the result that Dust Capacity that filtrator 1036 collects weighs by 2 collected at suction.
More than operation is to make ion generating device 1021 work, (just the material that such processing is obtained calls ion processing tick dust to make positive ion and negative ion act on the operation of dust, calling ion processing tick antigenicity substance) from the material that wherein extracts, in order to compare, also carry out except making ion generating device 1021 do not work, other operation operation same as described above fully, collect the tick dust (promptly this relatively the dust of usefulness call untreated tick dust, the material from wherein extraction is called untreated tick antigenicity substance).
In addition, as carrying out the employed device of this operation, except the device shown in above-mentioned Figure 16, can also use various devices, for example shown in Figure 17 (symbolic representation same section or the considerable part identical) with Figure 16, returnable 1025 is set substitutes the suction pump 1037 among Figure 16 and reclaim filtrator 1036, collect the dust that nature falls.
＜estimate by enzyme linked immunological absorption inhibition method
For the reactivity through the tick antigenicity substance of ion processing and undressed tick antigenicity substance and tick autopath's SERUM IgE is carried out quantitative evaluation, confirm by enzyme linked immunological absorption inhibition method.
Specifically be the tick antigen that from spread the tick dust that reclaims the back, extracts, put in the centrifugal separator (Centriprep YM-10), carry out centrifugal concentrating with the rotating speed of 2500rpm.Again this concentrate is put in the centrifugal separator (ULTRA FLEE-MC), carries out centrifugal concentrating with the 7000rpm rotating speed.Ion processing tick antigenicity substance and undressed tick antigenicity substance to concentrating carry out 11 5 times of dilutions repeatedly from protein concentration 7.66 μ g/ml.The antigenicity substance 50 μ l and the patients serum IgE 50 μ l of 10 times of dilutions with dilution mix respectively, 4 ℃ of one nights of following preincubate.
On adsorbing with 96 orifice plates (ELISA 96-orifice plate), enzyme linked immunological applies the tick antigenicity substance (also can be the material of not spraying) that is diluted to 1 μ g/ml with sodium bicarbonate buffer solution (Bicarbonate buffer), every hole 50 μ l, static placement 2 hours.With buffer solution (Washing Buffer) plate is carried out 3 washings with washing, apply sealing then with damping fluid (Blocking buffer) 300 μ l, one night of static placement under 4 ℃.
After one night of static placement, plate is carried out 4 washings, provide sample 50 μ l through preincubate, static placement 4 hours to every hole respectively.After plate carried out 5 washings, apply through (the biotin labeling anti human IgE that 3% skimmed milk+1%BSA)/PBST dilution is 1000 times, every hole 50 μ l, static placement 2.5 hours.
After the static placement, plate is carried out 3 washings, apply through (the alkali phosphatase enzyme mark streptavidin 50 μ l that 3% skimmed milk+1%BSA)/PBST dilution is 1000 times, at room temperature static placement 1.5 hours.To plate carry out 4 times the washing after, apply Attophos (registered trademark) substrate buffer solution, every hole 50 μ l are placed to color development under the shading state.(Cyto (registered trademark) FluorII) measures its fluorescence intensity by spectrophotometer.Wherein, reagent is so long as not explanation especially in advance can be used all ingredients same as described above.
When not making being untreated of ion generating device work (just undressed tick antigenicity substance) and make this element work, and make the space average concentration of both positive and negative ion reach 3000/cm respectively 3When handling under the condition tick antigenicity substance of ion processing (just through), the reactivity (associativity) of antigenicity substance and tick autopath's SERUM IgE antibody is studied.Its result as shown in figure 18.
As shown in figure 18, untreated tick antigenicity substance, its 50% inhibition (the tick antigenicity substance is reduced to 50% to the reactivity of SERUM IgE antibody) necessary tick antigenicity substance amount is 500ng/ml; In contrast, through the antigenicity substance of ion processing, it 50% suppresses necessary tick antigenicity substance amount and is 1000ng/ml at least, can confirm that it reacts inactivation rate is 74%.Wherein reacting inactivation rate can obtain by the chemical formula identical with above-mentioned chemical formula (1).
So just can confirm that not only facedown originality of both positive and negative ion material has an effect, and its effect also relates to the tick dust that contains antigenicity substance.And can confirm to reach 3000/cm respectively when the space average concentration of both positive and negative ion 3The time, can bring into play the effect that makes the antigenicity substance inactivation.
＜embodiment 5 〉
In the present embodiment, the space average concentration of both positive and negative ion is respectively 10000/cm 3(peak to peak value voltage is 100V between the electrode of this ion generating device 1021, and the air output of this fan blower 1033 is 8m 3/ minute), in addition other fully according to embodiment 4 identical operations, confirm of the effect of both positive and negative ion for the tick dust.The result as shown in figure 19.
As shown in figure 19, untreated tick antigenicity substance, its 60% inhibition (the tick antigenicity substance is reduced to 60% to the reactivity of SERUM IgE antibody) necessary tick antigenicity substance amount is 345ng/ml; In contrast, through the tick antigenicity substance of ion processing, the necessary tick antigenicity substance amount of its 60% inhibition is 3823ng/ml, can confirm that its reaction inactivation rate is 91%.Wherein reacting inactivation rate can obtain by aforementioned calculation formula (1) equally.
So just can confirm to be respectively 10000/cm when the space average concentration of both positive and negative ion 3The time, can bring into play the effect that makes the antigenicity substance inactivation.
If Figure 18 and Figure 19 are compared, though existing 50% suppresses and 60% difference that suppresses, but can think according to Figure 18 judge 50% suppress and reaction inactivation rate during 60% inhibition roughly the same, so can confirm, the high more reaction inactivation rate of space average concentration is high more.
If like this according to the inventive method, by making the both positive and negative ionization, can make the antigenicity substance inactivation effectively, so can expect effectively to reduce various allergic diseases such as the pollinosis that causes by this antigenicity substance and tick allergy.
Simultaneously, by at inner or outside the inventive method or the device of using of conditioner, can carry the air that makes the antigenicity substance inactivation and might directly make to be suspended in airborne antigenicity substance inactivation by the effect of emitting above-mentioned ion.
In above-mentioned each embodiment, stress anaphylactogen contained in pollen and the tick is illustrated, even can think for the allergen that is contained in mould etc. except that pollen and tick, be the air cleaning unit of foundation with the present invention, also can bring into play effect.
Current disclosed embodiment and embodiment are example in all respects, it should be considered as limiting.The scope of the invention is not above-mentioned explanation, but shown in claim, comprises the meaning and all modifications in its scope that equate with claim.
Practicality on the industry
As follows, the activated gas of the application of the invention makes method of evaluating performance and the device of antigenicity substance inactivation, can accurately and easily to various activated gas the performance of various antigenicity substance inactivations be estimated.

Claims (14)

1. activated gas makes the method for evaluating performance of antigenicity substance inactivation, it possesses reacts skyborne antigenicity substance of suspension and activated gas, obtain the step of processed antigen material, antibody and aforementioned processed antigen material with respect to aforementioned antigenicity substance are reacted, measure the step in conjunction with activity of aforementioned processed antigen material with respect to aforementioned antibody.
2. activated gas makes the method for evaluating performance of antigenicity substance inactivation, it possesses reacts skyborne antigenicity substance of suspension and activated gas, obtain the step of processed antigen material, antibody and aforementioned processed antigen material with respect to aforementioned antigenicity substance are reacted, measure the step of aforementioned processed antigen material with respect to aforementioned antibody binding activity, with aforementioned processed antigen material in conjunction with active and combine the step that activity compare of aforementioned antigenicity substance with respect to aforementioned antibody.
3. make the method for evaluating performance of antigenicity substance inactivation according to the activated gas described in the claim 1, wherein, the step that previous reaction is carried out is included in the container step of scattering the solution that contains aforementioned antigenicity substance, makes step that the solution that contains aforementioned antigenicity substance of aforementioned distribution suspends in aforementioned container and the step that imports aforementioned activated gas in aforementioned container.
4. make the method for evaluating performance of antigenicity substance inactivation according to the activated gas described in the claim 1, wherein, the step that obtains aforementioned processed antigen material comprises by aforementioned antigenicity substance being applied vibration and/or impacting, and makes the aforementioned antigenicity substance skyborne step that suspends.
5. make the method for evaluating performance of antigenicity substance inactivation according to the activated gas described in the claim 4, wherein, the aforementioned step that suspends comprises the step that aforementioned antigenicity substance is arranged on the step on the sample bench with flexibility and aforementioned sample bench is applied vibration and/or impacts.
6. make the method for evaluating performance of antigenicity substance inactivation according to the activated gas described in the claim 5, wherein, the aforementioned step that suspends comprise aforementioned antigenicity substance be arranged on the sample bench that one or more materials of being selected from bed clothes, woollen blanket, cushion, pillow, mat, sponge, cloth, paper, styrenic foams form with flexibility step and by beaing and/or vibrate aforementioned sample bench, the step that aforementioned sample bench is applied vibration and/or impacts.
7. make the method for evaluating performance of antigenicity substance inactivation according to the activated gas described in the claim 1, wherein, the step that obtains aforementioned processed antigen material comprises one or more the step of gas reaction that makes aforementioned antigenicity substance and be selected from the gas that contains positive ion, the gas that contains negative ion, the gas that contains free radical, ozone gas, nitric acid gas.
8. make the method for evaluating performance of antigenicity substance inactivation according to the activated gas described in the claim 1, wherein, the step that obtains aforementioned processed antigen material comprises and makes one or more material and the activated gas that is selected from antigenicity substance contained in Chinese fir pollen and/or the tick dust, Chinese fir pollen, tick dust react and obtain the step of processed antigen material.
9. make the method for evaluating performance of antigenicity substance inactivation according to the activated gas described in the claim 1, wherein, aforementioned determination step comprises by ELISA method and/or ELISA inhibition method, antibody and aforementioned processed antigen material with respect to aforementioned antigenicity substance are reacted, measure the step in conjunction with activity of aforementioned processed antigen material with respect to aforementioned antibody.
10. make the method for evaluating performance of antigenicity substance inactivation according to the activated gas described in the claim 1, wherein, aforementioned determination step comprises by except that the people and animal that have with respect to the antibody produced cell of aforementioned antigenicity substance are carried out intradermal reaction test and/or conjunctival reaction test, make aforementioned antibody and aforementioned processed antigen substance reaction, measure the step in conjunction with activity of aforementioned processed antigen material with respect to aforementioned antibody.
11. make the generating apparatus of processed antigen material of the performance evaluation sample of the skyborne antigenicity substance inactivation that suspends as activated gas, it possesses container, scatters the device of antigenicity substance and the device of aforementioned activated gas takes place or imports in aforementioned container in aforementioned container.
12. according to making the generating apparatus of processed antigen material of the performance evaluation sample of antigenicity substance inactivation as activated gas described in the claim 11, wherein, aforementioned container contains the partially transparent material or all is transparent material.
13. make the generating apparatus of processed antigen material of the performance evaluation sample of the skyborne antigenicity substance inactivation that suspends as activated gas, it comprises container, antigenicity substance is sealing into the device in the aforementioned container and the device of aforementioned activated gas takes place in aforementioned container or imports.
14. according to making the generating apparatus of processed antigen material of the performance evaluation sample of antigenicity substance inactivation as activated gas described in the claim 13, wherein, aforementioned container contains the partially transparent material or all is transparent material.
CNB2004800104221A 2003-02-18 2004-02-13 Method of evaluating performance of activation gas deactivating antigenic substance and apparatus for generating processed antigenic substance used as evaluation sample of the evaluating method Expired - Fee Related CN100510748C (en)

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Publication number Priority date Publication date Assignee Title
CN105039582A (en) * 2015-09-10 2015-11-11 浙江大学 Congenital and susceptible deafness gene detection kit

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN105039582A (en) * 2015-09-10 2015-11-11 浙江大学 Congenital and susceptible deafness gene detection kit

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