JP3806347B2 - Allergen deactivation method and air conditioner - Google Patents

Description

【００３１】
酸またはアルカリの存在する条件下で維持されたアレルゲン蛋白質はそのｐＨ条件の作用によって変性され、アレルゲンとしての活性を失う。
【００３２】
（３）酵素を保持させたアレルゲン不活性フィルター
本発明に従うフィルターの第３の例は、図３に示す通り、吸水性ポリマーの径より小さいメッシュを有した第１の支持体３１と、この第１の支持体３１の片側に配置された、吸水性ポリマーの径より小さいメッシュを有した第２の支持体３２と、前記第１の支持体と第２の支持体間に配置された吸水性ポリマー層３４とを具備する（図３）。
【００３３】
酵素の担体としては、アクリル酸−ビニルアルコール共重合体、アクリル酸−ナトリウム重合体などの吸水性ポリマーを使用し得る。また、前記担体の吸水能は少なくとも乾燥重量１ｇの吸水性ポリマー当たり６００ｇ程度であることが好ましい。
【００３４】
前記第１および第２の支持体としては、例えば、樹脂製のものが挙げられるが、これに限るものではない。また、前記吸水性ポリマー３４は、所定数のプロテアーゼを含むことが好ましい。酵素を保持させたアレルゲン不活性フィルター３５は、例えば、担体を織り込んだフィルターを作製した後に酵素溶液をフィルターに塗布してもよく、担体に酵素溶液を塗布した後にこれをフィルターに織り込んでもよい。また、相対湿度（即ち、水分活性）を７０％以下に保つことによってカビ汚染も減少できる。更に、界面活性剤（例えば、０．１％程度）を共存させることによって更なる効果も期待できる。
【００３５】
アレルギーはアレルゲンが直接皮膚に接触することによっても生じるが、その多くは、空気中の浮遊アレルゲンを吸引することによって生じる。従って、空気中のアレルゲンを排除できれば、アレルギー症状を緩和できると考えられる。上述のような本発明に従う装置によって空気中に浮遊するアレルゲンを不活性化することが可能であり、それにより住環境の大幅な改善が期待できる。
【００３６】
【実施例】
以下、例を用いて本発明を更に詳しく説明する。
【００３７】
１．方法および材料
（１）アレルゲンの検出
アレルゲンの存在を検出するためには、人間や動物が持っているアレルギー反応に類似する検出システムを採用することが望ましい。以下の試験では、本発明者らは、アレルギー反応が抗体抗原反応であることから、抗体を使った測定法を採用した。
【００３８】
（２）アレルゲン
以下の例において行った試験では、アレルゲンの１つとしてダニ抽出物を使用した。また、使用したダニ抽出物は、コスモバイオ社から入手したダニ抽出物−Ｄｆ（これはダニ虫体をリン酸緩衝液ですりつぶしその上清を凍結乾燥した物である）であり、この１０ｍｇを、処方箋に従って２．５ｍＬのＰＢＳ溶液に溶解した。得られた溶液を以下、ダニ原液と称し、使用時にはＰＢＳ溶液で適当な濃度に希釈して実験に用いた。
【００３９】
２．実施例
（１）方法
実施例１ アレルゲン測定１
本例では、ウエスタンブロッティング法（一般的にはドットブロット法とも称する）によってダニ抽出物中のダニアレルゲンの測定を行った。使用したダニ抽出物の量は蛋白質量として０．０１、０．０８３、０．２５、０．８３および２．５μｇである。
【００４０】
ダニ抽出物に対する抗体は、ダニ抽出物で免役したウサギの全血清（コスモバイオ社から入手した、使用時には２５００倍希釈した）を用いた。また、ウサギ全血清に対する抗体は、西洋わさびペルオキシダーゼで修飾したウサギＩｇに対する抗体（アマシャム社製）を用いた。使用前に１０００倍に希釈してから使用した。
【００４１】
アレルゲンの検出には、アマシャム社製のウエスタンブロッティング検出システムであるＥＣＬ＋Ｐｌｕｓを用い、製造元のマニュアルに従って行った。その他の操作手順は一般的な方法に準じて行った。
【００４２】
ダニ抽出物濃度に依存して得られた発光量から、ダニ抽出物中のダニアレルゲン量に関する検量線を取得した。図４はウエスタンブロッティングの結果を示す模式図である。図４から明かであるように得られた発光量は濃度依存的に増大した。従って、ダニ抽出物中の蛋白量はダニアレルゲン量を反映していることが明かになった。また、ウエスタンブロッティングにより得られた発光量を数値化して得た検量線を図５に示す。
【００４３】
実施例２ アレルゲン測定２
本例を含む以下の例では、サンプル中のダニアレルゲンの相対量をダニスキャンによって測定した。まず、ダニスキャンの定量性を確認するために、ダニ抽出物中のダニアレルゲンについての検量線を作製した。
【００４４】
前記ダニ原液をＰＢＳ溶液で希釈した溶液について、ダニアレルゲン測定システムとしてダニスキャン（DaniScan、登録商標、アサヒビール薬品社製）を用いてアレルゲンを検出した。
【００４５】
ダニスキャンは、抗原抗体反応を利用して簡易にダニアレルゲンを検出できるキットである。図６に示すダニスキャンに具備されるちり採取器１のサンプル添加部２に被検サンプルを添加し、そこに現像液を添加すれば、被検サンプル中のダニアレルゲンの量を反映したバンド３が得られる。前記バンドから被検サンプル中の相対的なアレルゲンを検出することが可能である。本明細書において、ダニスキャンの各バンドは、便宜上「太実線」、「中太実線」「細実線」、「破線」および「表示なし」で示し、これらの表示は、「太実線」＞「中太実線」＞「細実線」＞「破線」＞「表示なし」の順で４段階の発色強度を模式的に示している。
【００４６】
また、サンプル中に含まれるダニアレルゲン量は、ダニ抽出物中のダニ抗原に結合したダニ抗体に由来する発色強度のピーク値（Ｔ）とダニ抗原に結合しなかったダニ抗体に由来する発色強度のピーク値（Ｃ）から、以下の式に従ってピーク比を求めることによって評価した。図７にダニスキャン上の位置と発色強度の関係を示すグラフを示す。ここで前記ピーク値（Ｔ）とピーク値（Ｃ）は図７中の「Ｃ」と「Ｔ」のピークに相当する。
【００４７】
発色強度からピークＣとピークＴの発色強度からピーク比を求めるための式は；
ピーク比（％）＝Ｔ／（Ｔ＋Ｃ）＊１００
である。このとき、Ｃ＞Ｔであればサンプル中のダニアレルゲン量は少なく、Ｔ＞Ｃであればサンプル中にダニアレルゲンが大量に存在していることを示す。
【００４８】
図６に、ダニアレルゲン陽性（即ち、アレルゲンが多く存在する場合）のバンドが示されたダニスキャンの模式図を示す。Ｃの位置のバンドはダニ抗原に結合しなかったダニ抗体に由来するバンドであり、Ｔの位置のバンドはダニ抗原に結合したダニ抗体に由来するバンドである。図８にダニ抽出物中に存在するダニアレルゲンに関する検量線を示す。
【００４９】
実施例３ 熱によるアレルゲンの不活性化
熱処理によるダニアレルゲン不活性化作用を証明するために以下の実験を行った。
【００５０】
ダニ原液のＰＢＳ溶液による１０倍希釈液を調製した。得られた１０倍希釈液を７本のマイクロチューブに分注し、そのうちの１本を加熱時間０分の対照区とした。残りの６本をヒートブロック上で８０℃に加熱し、それぞれ、１０、２０、３０、４０、５０および６０分間維持した。これを処理区とした。加熱終了後、氷上にて冷却した。その後、各サンプルからそれぞれ５μＬずつを抜き取ってダニスキャンに添加してアレルゲンの検出を行った。添加した５μＬには蛋白質として２μｇのダニ抽出物が含まれていることになるので、１ダニスキャン当たり２μｇのダニ抽出物を添加したことになる。
【００５１】
熱処理時間とサンプル中のダニアレルゲン量の関係を示す結果を図９のグラフに示す。図９のグラフの縦軸には、対照区におけるピーク比を１００とした、処理区における相対ピーク比を示し、横軸には熱処理時間を示す。
【００５２】
前記グラフから明かであるように、熱処理によりアレルゲンは不活性化された。特に加熱処理１時間後にはかなりのアレルゲンが不活性化した（図９）。
【００５３】
実施例４ アルカリによる不活性化
アルカリ処理によるダニアレルゲン不活性化作用を証明するために以下の実験を行った。
【００５４】
まず、氷上で５本のマイクロチューブ内で、それぞれダニ原液５０μＬと５Ｍ水酸化ナトリウム５０μＬを混合した。得られた混合液の水酸化ナトリウムの終濃度は２．５Ｍである。それらのマイクロチューブを４０℃と６０℃の温度条件下で、それぞれ０、１０および３０分間維持した。反応終了後、５Ｍ塩酸５０μＬを加えて氷上で中和した。その後、得られた反応液の各１μＬ（蛋白質として１．３３μｇダニ抽出物／ダニスキャン）をダニスキャンに添加し、ダニアレルゲンを検出した。図１０に結果を示す。図１０は反応後のダニスキャンに見られるバンドを模式的に示した図である。図１０の１は処理なしから得た結果（即ち、１ダニスキャン当たり１．３３μｇのダニ抽出物のみを添加した結果）であり、これは対照サンプルである。図１０の２から４は、ダニ抽出物について４０℃でそれぞれ０分、１０分および３０分間に亘ってアルカリ処理を行った結果である。図１０の５および６は、ダニ抽出物について、６０℃で、それぞれ０分および１０分間アルカリ処理した場合の結果である。図１０から明かであるように４０℃および６０℃ともに、１０分間の処理でアレルゲンが不活性化した（図１０）。
【００５５】
実施例５ 酸による不活性化
酸処理によるダニアレルゲン不活性化作用を証明するために以下の実験を行った。
【００５６】
まず、４本のマイクロチューブにダニ原液５０μＬと５Ｍ塩酸５０μＬを氷上で混合した。得られた混合液の塩酸の終濃度は２．５Ｍである。それらのマイクロチューブを６０℃の温度条件下で、それぞれ０、１０、３０および６０分間維持した。反応終了後、５Ｍの水酸化ナトリウム５０μＬを加えて氷上で中和した。得られた反応液の各１μＬ（蛋白質として０．１３３μｇダニ抽出物／ダニスキャン）をダニスキャンに添加し、ダニアレルゲン量を測定した。
【００５７】
図１１に結果を示す。図１０は反応後のダニスキャンに見られるバンドを模式的に示した図である。図１１の１は処理なし（即ち、１ダニスキャン当たり蛋白質として１．３３μｇのダニ抽出物を添加した）であり、これを対照サンプルとした。図１１の２から５は、ダニ抽出物を６０℃でそれぞれ０分、１０分、３０分および６０分間酸処理した場合の結果を示す。図１１から明かであるように６０℃で６０分間の酸処理により大部分のアレルゲンが不活性化されたことが確認された。
【００５８】
実施例６ プロテアーゼによる不活性化１
プロテアーゼによるダニアレルゲン不活性化作用を証明するために以下の実験を行った。
【００５９】
まず、３本のマイクロチューブのそれぞれに、ダニ原液９０μＬとｐｆｕ プロテアーゼＳ（以下、ｐｆｕと記す、宝酒造社製）の８μＬ（０．８ユニット）を氷上で加えた。これらのうち１本を０℃で１０分、他の１本を９５℃で１０分間、残りの１本を８０℃で１０分の加熱を行った。酵素添加なしの対照サンプルについては９５℃で１０分間の加熱を行った。これらの熱処理後、全てのマイクロチューブを氷上に戻し、プロテアーゼ阻害剤であるフェニルメチルスルホニルフルオリド（以下、ＰＭＳＦと記す）溶液を加えた（ＰＭＳＦの終濃度は４ｍＭ）。
【００６０】
その後、ＰＭＳＦを加えた各サンプルから５μＬを抜き取ってダニスキャンに添加してダニアレルゲンを検出した。このとき１ダニスキャン当たり蛋白量として１８μｇのダニ抽出物が含まれる。
【００６１】
結果は図１２に示した。図１２の１は酵素添加し、０℃で１０分間反応したサンプル、図１２の２は酵素なしで９５℃１０分間維持した対照サンプル、３は酵素添加で９５℃で１０分間処理したサンプル、４は酵素添加で８０℃１０分間処理したサンプルについての結果を示す。図から明かであるように、上述の至適温度でのプロテアーゼ処理を１０分間行えば、ダニアレルゲン活性は失活できる。
【００６２】
実施例７ プロテアーゼによる不活性化２
次にプロテアーゼと尿素を併用した場合のダニアレルゲン不活性化作用を証明するための実験を行った。
【００６３】
３本のマイクロチューブのそれぞれに、ダニ原液２０μＬとｐｆｕの３．２μＬ（０．３２ユニット）を氷上で加え、更にＰＢＳ溶液で調製した１０Ｍの尿素の１６．８μＬを加えた。尿素の最終濃度は４．２Ｍである。これらを次に６０℃で０、１０および３０分間加熱した。加熱後、氷上に戻し反応を終了した。得られたサンプルからそれぞれ１μＬを抜き取ってダニスキャンによりアレルゲンを検出した。このとき、１つのダニスキャンに添加したダニ抽出物は蛋白量として２μｇである。この結果は図１３の１から３に示す。図１３の１、２および３は、それぞれ、６０℃で０、１０および３０分間加熱したサンプルについての結果である。
【００６４】
更に以下の試験を同様に行った。４本のマイクロチューブのそれぞれに、ダニ原液１２μＬとｐｆｕの８μＬ（０．８ユニット）を氷上で加え、更に１０Ｍの尿素を８０μＬを加えた。尿素の最終濃度は８Ｍである。これらを次に４０℃で０、１０、３０および６０分間加熱した。得られたサンプルからそれぞれ１μＬを抜き取ってダニスキャンによりアレルゲン量を測定した。このとき、１つのダニスキャンに添加したダニ抽出物は蛋白量として０．４８μｇである。この結果は図１３の５から８に示す。図１３の５、６、７および８は、それぞれ、４０℃で０、１０、３０および６０分間加熱したサンプルについての結果である。
【００６５】
また、対照サンプルとして、酵素および加熱処理なしのサンプルを調製し試験した。氷上で、１本のマイクロチューブにダニ原液１２μＬ、１０Ｍの尿素８０μＬおよびＰＢＳ溶液８μＬを加えた。尿素の最終濃度は８Ｍである。このマイクロチューブには加熱を行わなかった。このサンプルから１μＬを抜き取ってダニスキャンによりアレルゲンを検出した。このとき、１つのダニスキャンに添加したダニ抽出物は蛋白量として０．４８μｇである。この結果は図１３の４に示す。
【００６６】
図１３に示す結果から明かであるように、尿素４．２Ｍ存在下で６０℃で加熱した場合１０分後にはアレルゲンは不活性化された。また、尿素８Ｍ存在下で４０℃で加熱した場合には３０分後にはアレルゲンは不活性化された。
【００６７】
実施例８ プロテアーゼによる不活性化３
更に、もう１つのプロテアーゼの実施例としてパパインを使用した場合のダニアレルゲン不活性化作用を証明するために実験を行った。
【００６８】
まず、１０Ｍ尿素溶液４００μＬに０．２ｇ／ｍＬのパパイン（ナガセケムテックス社製、食品用精製パパイン）とＰＢＳ溶液１００μＬを混合して８Ｍの尿素パパイン溶液を調製した。
【００６９】
８本のマイクロチューブのそれぞれにダニ原液２０μＬと８０μＬの８Ｍ尿素パパイン溶液を添加した。尿素の最終濃度は６．４Ｍである。これらについて４０℃での加熱を０、１０、３０、６０、１２０、１６５、２８０および３２０分間行った。前記加熱の後、各サンプルからそれぞれ１８μＬずつ取り出し、それぞれ１０ｍＭのアンチパイン（antipain, BACHEM社製）の２μＬずつを加えた。アンチパインの最終濃度は１ｍＭである。
【００７０】
得られた溶液のそれぞれ１μＬずつ採取してダニスキャンに添加し、ダニアレルゲンを検出した。このとき１ダニスキャン当たりには蛋白質として０．７２μｇのダニ抽出物を添加されたことになる。
【００７１】
得られた結果は、図１４の１から８に示した。図１４の１、２、３、４、５、６、７および８は、それぞれ４０℃で０、１０、３０、６０、１２０、１６５、２８０および３２０分間行って得たサンプルについての結果である。これらの結果から、上記の条件では１６５分の加熱でアレルゲンの不活性化が可能であり、特に３２０分の加熱では殆どのアレルゲンが不活性化された。
【００７２】
更に、３０℃で加熱した場合のパパインの効果を試験した。
【００７３】
５本のマイクロチューブのそれぞれにダニ原液２０μＬと１８０μＬの８Ｍ尿素パパイン溶液を添加した。尿素の最終濃度は７．２Ｍである。これらについて３０℃での加熱を０、１０、３０、１２０および１８０分間行った。前記加熱の後、各サンプルからそれぞれ１８μＬずつ取り出し、それぞれ１０ｍＭのアンチパイン（antipain, BACHEM社製）の２μＬずつを加えた。アンチパインの最終濃度は１ｍＭである。
【００７４】
得られた溶液のそれぞれ１μＬずつをダニスキャンに添加し、ダニアレルゲンを検出した。このとき１ダニスキャン当たりには蛋白質として０．３６μｇのダニ抽出物が添加されたことになる。
【００７５】
得られた結果は、図１４の９から１３に示した。図１４の９、１０、１１、１２および１３は、それぞれ３０℃で０、１０、３０、１２０および１８０分間行って得たサンプルについての結果である。上記の条件では１２０分の加熱でアレルゲンの不活性化が可能であった（図１４）。また、図１４には示していないが、上記の条件では３２０分の加熱により殆どのアレルゲンが不活性化できた。
【００７６】
以上の実施例からも明かであるように、本発明に従うと、蛋白質がエピトープとなるアレルゲンの不活性化方法として、普遍的に使える解決方法が提供される。また、本発明は蛋白質がエピトープとなるアレルゲンに関して、全てに対応できるものである。
【００７７】
（２）フィルターおよび空調の実施例
以下に、本発明に従うフィルターおよび空調機の実施例を図面を参照して説明する。
【００７８】
実施例９
図１を参照しながら実施例９を説明する。図中の符番１１は、アレルゲンをトラップする不織布フィルターを示す。この不織布フィルター１１の下面には、花粉粒子（２０〜３０μｍ）やダニ（特にダニの糞、１０〜４０μｍ）の径より小さいメッシュを有したステンレス製の繊維状面発熱体１２が配置されている。このステンレス製の線維状面発熱体（商品名：ソフテレック、帝人社製）１２の下部には、前記面発熱体１２を加熱する電熱ヒータ１３が配置されている。
【００７９】
このように実施例９に係るアレルゲン不活性フィルター１４は、アレルゲンをトラップする不織布フィルター１１と、この不織布フィルター１１の下面に配置された、花粉粒子やダニの径より小さいメッシュを有したステンレス製面発熱体１２と、この繊維状面発熱体１２の下部に配置された、該面発熱体１２を加熱する電熱ヒータ１３とを具備した構成となっている。
【００８０】
こうしたアレルゲン不活性フィルター１４は、後述する空調機（図１５参照）の所定の位置にエアコン搭載可能な大きさ（例えば、５ｃｍ×１０ｃｍ）で取り付けられて使用される。空調機の空気取り込み口にアレルゲン不活性フィルター１４を取り付ける際は、不織布フィルター１１をアレルゲン含有空気（矢印Ａ）の入口側に配置する。そして、空調機がＯＦＦの時、フィルター１４の電熱ヒータ１３をＯＮにして面発熱体１２の加熱温度を例えば約７０℃で発熱される。
【００８１】
これにより、アレルゲン含有空気は不織布フィルター１１を通過するが、高温状態に保持された面発熱体１２で花粉粒子やダニが捕獲され、ここでアレルゲン蛋白質の変性が行われ、アレルゲンとしての活性は失われ、ステンレス面発熱体１２からはアレルゲン軽減空気（矢印Ｂ）が通過する。
【００８２】
従って、実施例１のアレルゲン不活性フィルター１４によれば、従来のように害虫駆除用の薬品を用いることがないので、薬品自体の人体への影響を回避できるとともに、前記発熱体を所定の温度で加熱するだけで自動的にアレルゲン量を軽減することができる。
【００８３】
実施例１０
図２を参照する。実施例１０に係るアレルゲン不活性フィルター２１は、不織布フィルター２２に、強酸性陽イオン交換樹脂（図示せず）を保持させた構成となっている。ここで、強酸性陽イオン交換樹脂は、Ｒ−ＳＯ３・Ｈ（但し、Ｒは高分子基体を示す）であり、酸を使って再生が可能である。この強酸性陽イオン交換樹脂は、使用前に活性化しておく。
【００８４】
実施例１０によれば、アレルゲン含有空気Ａが不織布フィルター２２を通過するだけで該不織布フィルター２２に保持された強酸性陽イオン交換樹脂によりアレルゲン蛋白質はそのｐＨの影響で変性し、アレルゲンとしての活性を失う。従って、不織布フィルター２２からはアレルゲン軽減空気Ｂが排気される。従って、実施例１０のアレルゲン不活性フィルター２１によれば、従来のように害虫駆除用の薬品を用いることがないので薬品自体の人体への影響を回避できるとともに、強酸性陽イオン交換樹脂を保持させた不織布フィルター２２にアレルゲン含有空気Ａを通すだけで自動的にアレルゲン量を軽減することができる。
【００８５】
なお、実施例１０では、強酸性陽イオン交換樹脂を使用する場合について述べたが、これに限らず、上記化合物１に示す強塩基性陰イオン交換樹脂を使用してもよい。但し、この場合も、強塩基性陰イオン交換樹脂は使用前に活性化しておくことが好ましい。
【００８６】
実施例１１
図３を参照して実施例１１を説明する。図中の符番３１は吸水性ポリマー３２が移動しない大きさで且つ花粉粒子やダニを通すメッシュ（＞５０μｍ）を有した樹脂製の第１の支持体を示す。この第１の支持体３１の下側には吸水性ポリマー３４が移動しない大きさで且つ花粉粒子やダニを通さないメッシュ（＞５０μｍ）を有した樹脂製の第２の支持体３３が配置されている。前記第１の支持体３１と第２の支持体３３の間には、吸水性ポリマー３２からなる吸水性ポリマー層３４が挟まれて配置されている。ここで、吸水性ポリマー３２は、例えば、５０万ユニット／フィルターのプロテアーゼを含んでいる。前記吸水ポリマー層３４は、例えば、プロテアーゼを含む酸素溶液にポリマーを含浸させることにより形成することができる。
【００８７】
実施例１１に係るアレルゲン不活性フィルター３５は、樹脂製の第１の支持体３１と第２の支持体３３の間に吸水性ポリマー層３４を挟んだ構造を有する。それにより、アレルゲン含有空気Ａが吸水性ポリマー層３４を通過するだけで、蛋白質を分解する酵素によってアレルゲンが分解される。従って、実施例１１のアレルゲン不活性フィルター３５によれば、従来のように害虫駆除用の薬品を用いることがないので薬品自体の人体への影響を回避できるとともに、吸水性ポリマー層３４にアレルゲン含有空気Ａを通すだけで自動的にアレルゲン量を軽減することができる。
【００８８】
実施例１２
図１５は、実施例９から１１に係るアレルゲン不活性フィルターをエアコンの空気取り込み口に取り付けた例を示す。図１５において、符番４２はエアコン用冷却ユニットを示し、符番４３は筐体を示す。この筐体４３の内側には、ファン４４などが配置されている。ここで、前記アレルゲン不活性フィルター１４は、該フィルター１４の主要な構成である不織布フィルター２１が空気アレルゲン含有空気の入口に位置するよう配置される。
【００８９】
このように実施例１２に係る空調機は、エアコンの空気取り込み口にアレルゲン不活性フィルター１４を配置した構成になっている。そのため、空調機がＯＦＦの時、フィルター１４の電熱ヒータ１３をＯＮにして面発熱体１２を発熱させることにより、高温状態に保持された面発熱体１２で花粉粒子やダニが捕獲され、ここでアレルゲン蛋白質の変性が起こり、アレルゲンとしての活性が失われる。ステンレス面発熱体１２からはアレルゲン軽減空気が通過する。なお、前記電熱ヒータ１３には空調機の電源を利用する。
【００９０】
エアコン、空気清浄機などの空調機に、本発明のアレルゲン分解機能を付加することで住環境の著しい改善が図れる。
【００９１】
【発明の効果】
本発明に従うと、アレルゲンを特異的に排除する方法が提供される。
【図面の簡単な説明】
【図１】本発明に従うアレルゲン不活性フィルターの例を示す図。
【図２】本発明に従うアレルゲン不活性フィルターの例を示す図。
【図３】本発明に従うアレルゲン不活性フィルターの例を示す図。
【図４】ウエスタンブロッティングの結果の模式図。
【図５】ウエスタンブロッティングにより得た検量線を示すグラフ。
【図６】サンプル中にダニアレルゲンが多量に存在する場合に得られるバンドが示されたダニスキャンの模式図。
【図７】ダニスキャン上のバンドの位置と発色強度の関係を示すグラフ。
【図８】ダニ抽出物中に存在するダニアレルゲンに関する検量線を示すグラフ。
【図９】熱処理時間とサンプル中のダニアレルゲン量の関係を示すグラフ。
【図１０】反応後のダニスキャンに見られるバンドを模式的に示した図
【図１１】反応後のダニスキャンに見られるバンドを模式的に示した図
【図１２】反応後のダニスキャンに見られるバンドを模式的に示した図
【図１３】反応後のダニスキャンに見られるバンドを模式的に示した図
【図１４】反応後のダニスキャンに見られるバンドを模式的に示した図
【図１５】図１のアレルゲン不活性フィルターを空気の入口側に取り付けた本発明に係る空調機を示す図。
【符号の説明】
１．ちり採取器 ２．サンプル添加部 ３．バンド １１.不織布フィルター １２．繊維状面発熱体 １３．電熱ヒータ １４．アレルゲン不活性フィルター ２１、２２．不織布フィルター ３１．第１の支持体 ３２．吸水性ポリマー ３３．第２の支持体 ３４．吸水性ポリマー層 ３５．アレルゲン不活性フィルター ４２．エアコン用冷却ユニット ４３．筐体 ４４．ファン[0001]
BACKGROUND OF THE INVENTION
It relates to a method for inactivating allergens.
[0002]
[Prior art]
In recent years, allergy issues have been highlighted. Cedar pollen is famous as an allergen, but due to recent housing circumstances such as high airtightness of houses, allergies due to pests such as mites and their excreta are becoming serious. As a countermeasure against such allergies, various products such as cleaning tools for collectively processing house dust such as a vacuum cleaner and a mop have been sent to the world.
[0003]
Moreover, there are the following methods for protecting yourself from allergies that are generally performed, that is, methods for eliminating allergens. For example, in the case of pollen typified by cedar, a method of preventing pollen from being taken into the body by putting a fine mesh that does not allow pollen to pass through the mask worn when going out is also a typical example of a dust collector indoors. The method of catching them by the device to be performed is performed. However, even if pollen present in the living space is positively removed by a dust collector or the like, it is only captured as part of airborne matter. Therefore, such a method is not an aggressive allergen removal method.
[0004]
On the other hand, in the case of allergens derived from pests such as mites, the amount of pests is reduced by frequently cleaning bedding, sofas, tatami mats, floors, etc., thereby reducing the allergen amount. Currently. However, in such a method, although the amount of allergen temporarily decreases, the amount of allergen increases if the pests propagate. Therefore, considerable human power is required to maintain the allergen amount at a sufficiently low value. Although it is possible to control pests using chemicals, it is necessary to consider the influence of the chemicals themselves used on the human body, which is not a sufficient measure.
[0005]
[Problems to be solved by the invention]
In view of such circumstances, the object of the present invention is to specifically exclude allergens. Allergen inactivation Method And air conditioner Is to propose.
[0006]
[Means for Solving the Problems]
As a result of intensive studies for solving the above problems, the present inventors have found the following means. That is, (1 ) Inactivate the allergen by maintaining it in the presence of rotase and urea Allergen inactivation characterized by Method,( 2 ) Also characterized by the presence of heat ( 1 ) No Allerge N Activity Conversion Law and ( 3 (1) Or ( 2) Provided with an allergen inactive portion for performing the method for inactivating the allergen described It is characterized by It is an air conditioner.
[0007]
DETAILED DESCRIPTION OF THE INVENTION
1. Allergen inactivation method
Allergens that cause allergies are mainly composed of biological proteins. The present invention is based on the fact that we have made it possible to specifically and effectively inactivate allergens by denaturing proteins. Therefore, this specification reports for the first time that allergens can be inactivated by protein denaturation. The term “allergen” or “allergen substance” as used herein refers to a substance having activity as an allergen.
[0008]
According to the present invention, there is provided a method for inactivating an allergen by maintaining the allergen under the presence of one condition selected from the group consisting of heat, alkali, acid and protease.
[0009]
Conventionally, it is generally known that proteins are degraded by chemical denaturation with acid or alkali, physical denaturation at high temperature, and biochemical denaturation with proteolytic enzymes. However, it has not been reported that the property that can be an epitope that a protein may have, that is, its activity as an allergen, can be specifically inactivated by such protein modification, and it is understood by knowledge of biochemistry. No possible and effective allergen inactivation method has been reported so far. It is epoch-making that the present inventors have revealed that allergens can be inactivated by protein denaturation.
[0010]
Hereinafter, a method for inactivating allergen using heat, alkali, acid and protease will be described.
[0011]
(1) Heat
In accordance with the present invention, when allergens are inactivated under heat, for example, the allergens may be exposed to heat for about 60 minutes at temperatures above about 80 ° C. Even at a temperature of 80 ° C. or lower, for example, if the heating time is increased, the allergen can be inactivated.
[0012]
In one aspect of the present invention, the present invention has been accomplished on the basis of a unique and new idea of using heat to inactivate allergens. Therefore, one of the important points in the present invention resides in inactivating the allergen using heat, not conditions such as temperature and heating time. In other words, the present invention includes possible combinations of conditions such as a combination of temperature and heating temperature necessary for inactivating the allergen by heat.
[0013]
The term “conditions in which heat is present” used herein means that the allergen or an object containing the same is heated using a heat source known per se or indirectly through a heated medium. It can also be obtained by heating the object to be contained.
[0014]
(2) Alkali
When inactivating the allergen in the presence of alkali according to the present invention, for example, in the presence of about 2.5 M NaOH, or in the presence of a basic substance that can achieve similar conditions. And allergens may be present. In the present invention, the basic substance that can be used for obtaining the conditions in which alkali is present may be any substance that generates hydroxide ions when dissolved in water. For example, but not limited to, various salts such as sodium hydroxide and potassium hydroxide, and substances such as strongly basic anion exchange resins may be used. Further, in addition to alkaline conditions, in the presence of heat, inactivation can be performed efficiently. When treating with a combination of alkali and heat, by adjusting the heating temperature and heating time, a basic substance having a lower concentration than the above-mentioned conditions or a weakly basic substance having a lower alkalinity than the above-mentioned strong base Good allergen inactivation can also be achieved by using.
[0015]
In one aspect of the present invention, the present invention has been accomplished on the basis of a unique and new idea of using an alkali for allergen inactivation. Therefore, one of the important points in the present invention is not to be in the detailed condition regarding the presence of alkali, but to inactivate the allergen using alkali. In other words, the present invention includes all possible combinations of conditions relating to the inactivation of allergens by alkali and the concentrations of basic substances and the presence / absence of heating, heating temperature and time.
[0016]
(3) Acid
In accordance with the present invention, inactivation of allergens in the presence of acid includes, but is not limited to, for example, conditions in which HCl is present at a concentration of about 2.5 M, or similar conditions. The allergen may be present under conditions where an acidic substance capable of achieving the above is present. In the present invention, the substance that can be used for obtaining the condition in which an acid is present may be any substance that generates hydrogen ions when dissolved in water. Examples include, but are not limited to, acids such as hydrochloric acid and sulfuric acid, and strong acid cation exchange resins. Further, in addition to acid conditions, in the presence of heat, it is possible to inactivate efficiently. When treating with a combination of acid and heat, by adjusting the heating temperature and heating time, use an acidic substance having a lower concentration than the above-mentioned conditions or a weakly acidic substance having a weaker acidity than the above-mentioned strong acid. Also good allergen inactivation can be achieved.
[0017]
In one aspect of the present invention described above, the present invention has been achieved based on the unique and new idea of using acids for inactivation of allergens. Therefore, one of the important points in the present invention is not in the detailed conditions regarding the presence of acid, but in the use of acid to inactivate the allergen itself. In other words, the present invention includes all possible combinations of conditions relating to the inactivation of allergens by acid and the concentration of acidic substances, the presence or absence of heating, and the heating temperature.
[0018]
(4) Protease
As used herein, the term “protease” generally refers to enzymes that have the property of degrading proteins and peptides. The enzyme that can be used in the present invention may be any known acidic, neutral or basic protease. For example, it may be a serine protease such as trypsin, a cysteine protease such as papain, calpain, cathepsin B and cathepsin L, a protease such as aspartic protease such as pepsin, renin and cathepsin D and a metalloprotease. In addition, when the allergen is inactivated by a protease, the practitioner may arbitrarily select conditions such as temperature and coenzyme in consideration of the optimum conditions of the protease to be used.
[0019]
In addition, when allergens are inactivated by the prosthesis according to the present invention, it is possible to inactivate more efficiently if urea is present in addition to the protease used.
[0020]
For example, the concentration of the protease used in the present invention is about 1 unit per 0.45 μg of allergen protein, for example, in the case of pfu. Moreover, when using protease, the effect obtained changes with process temperature, process time, and the presence or absence of urea. Therefore, the concentration of the protease used is not limited to the above, and can be effectively used under various conditions.
[0021]
In one aspect of the present invention described above, the present invention has been achieved based on the unique and new idea of using proteases and proteases and urea in combination for inactivation of allergens. Therefore, one of the important points in the present invention is that the allergens are inactivated by utilizing protease and combined treatment of protease and urea, rather than the detailed conditions such as their concentration and treatment time and heating temperature and heating time. Is in the process itself. In other words, all possible combinations of conditions such as the type and concentration of the enzyme involved in the inactivation of the allergen by the prosthesis and urea, the presence or absence of urea and the concentration of urea, and the presence or absence of heating, and the heating temperature and time are included in the present invention. Are all included.
[0022]
2. Use for air conditioners
According to a further aspect of the present invention, the method described above can be used for an air conditioner. That is, the present invention comprises an allergen inactive portion for performing a method for inactivating an allergen by maintaining the allergen under the presence of one selected from the group consisting of heat, alkali, acid and protease. It also provides an air conditioner.
[0023]
The allergen inactive part used in accordance with the present invention may be any means capable of maintaining the allergen under the presence of one condition selected from the group consisting of heat, alkali, acid and protease. In the case where the allergen is maintained in the presence of heat, any known heating body may be used. When maintaining allergens in the presence of alkalis, acids or proteases, for example, common chromatographic techniques can be used. In that case, a compound or enzyme capable of providing an alkali and an acid may be immobilized on a carrier such as a filter or a particle, and the allergen contained in the mobile phase may be treated.
[0024]
Examples of filters that can be used according to the present invention are given below.
[0025]
(1) Allergen inert filter by heat
As shown in FIG. 1, the first example of the filter according to the present invention is a nonwoven fabric filter 11 for trapping allergens, a stainless steel surface heating element 12 disposed on one surface of the nonwoven fabric filter 11, and the stainless steel surface heating element. And a heater 13 for heating 12 (FIG. 1).
[0026]
The stainless steel surface heating element 12 is not limited to this, but includes, for example, a stainless steel fiber knitted mesh. As for the power source of the heater 13, since the allergen inert filter 14 is normally used by being incorporated in an air conditioner, it is preferable to use the power source of the air conditioner. The heater 13 is normally heated for a predetermined temperature and time when the air conditioner is OFF, but is heated at a high temperature that does not damage the components such as the air conditioner for a short time, and is heated at a low temperature for a long time. To do. However, the temperature and time can be appropriately set according to the material of the component. The allergen protein is denatured by heating the surface heating element 12 by the heater 13, and the activity as an allergen is lost.
[0027]
(2) Allergen inactive filter retaining acid or alkali
As shown in FIG. 2, the second example of the filter according to the present invention is characterized in that the non-woven fabric filter 21 holds a strong acidic cation exchange resin or a strong basic anion exchange resin (FIG. 2). Such a filter can be manufactured by weaving an ion exchange resin (for example, which may be a fiber or a granular resin) as shown below into a general filter.
[0028]
i) Strongly acidic cation exchange resin
In the present invention, for example, the general formula R-SO ₃ A strongly acidic cation exchange resin represented by H (where R is a polymer substrate) can be used. This resin can be regenerated using acid. It is necessary to activate with an acid before use.
[0029]
ii) Strongly basic anion exchange resin
In the present invention, for example, a strongly basic anion exchange resin represented by the following general formula can be used. This resin can be regenerated using a base. Before use, it is necessary to activate with a base.
[0030]
[Chemical 1]

[0031]
Allergen proteins maintained in the presence of acid or alkali are denatured by the action of the pH conditions and lose their activity as allergens.
[0032]
(3) Allergen inactive filter with enzyme retained
As shown in FIG. 3, the third example of the filter according to the present invention is disposed on one side of the first support 31 having a mesh smaller than the diameter of the water-absorbing polymer, and the first support 31. A second support 32 having a mesh smaller than the diameter of the water-absorbing polymer, and a water-absorbing polymer layer 34 disposed between the first support and the second support are provided (FIG. 3).
[0033]
As the enzyme carrier, a water-absorbing polymer such as an acrylic acid-vinyl alcohol copolymer and an acrylic acid-sodium polymer can be used. The water absorption capacity of the carrier is preferably at least about 600 g per water-absorbing polymer having a dry weight of 1 g.
[0034]
Examples of the first and second supports include those made of resin, but are not limited thereto. The water-absorbing polymer 34 preferably contains a predetermined number of proteases. For the allergen-inactive filter 35 holding the enzyme, for example, after preparing a filter woven with a carrier, the enzyme solution may be applied to the filter, or after applying the enzyme solution to the carrier, this may be woven into the filter. Also, mold contamination can be reduced by keeping the relative humidity (ie water activity) below 70%. Furthermore, a further effect can be expected by coexisting a surfactant (for example, about 0.1%).
[0035]
Allergies can also be caused by direct contact of the allergen with the skin, most of which is caused by inhalation of airborne allergens in the air. Therefore, it is thought that allergic symptoms can be alleviated if allergens in the air can be eliminated. The apparatus according to the present invention as described above can inactivate allergens floating in the air, and thus a great improvement in the living environment can be expected.
[0036]
【Example】
Hereinafter, the present invention will be described in more detail with reference to examples.
[0037]
1. Methods and materials
(1) Allergen detection
In order to detect the presence of allergens, it is desirable to employ a detection system similar to allergic reactions that humans and animals have. In the following tests, the present inventors adopted a measurement method using an antibody because the allergic reaction is an antibody-antigen reaction.
[0038]
(2) Allergen
In the tests performed in the following examples, mite extract was used as one of the allergens. The mite extract used was tick extract-Df obtained from Cosmo Bio Co. (this is a mite body that has been crushed with a phosphate buffer and its supernatant lyophilized). According to prescription, dissolved in 2.5 mL PBS solution. The obtained solution is hereinafter referred to as a tick stock solution, and was used in the experiment after being diluted to an appropriate concentration with a PBS solution at the time of use.
[0039]
2. Example
(1) Method
Example 1 Allergen measurement 1
In this example, mite allergens in mite extracts were measured by Western blotting (generally also referred to as dot blotting). The amount of mite extract used is 0.01, 0.083, 0.25, 0.83 and 2.5 μg as protein mass.
[0040]
The antibody against the mite extract was the whole serum of rabbits immunized with the mite extract (obtained from Cosmo Bio, diluted 2500 times when used). As an antibody against rabbit whole serum, an antibody against rabbit Ig modified with horseradish peroxidase (Amersham) was used. Before use, it was used after diluting 1000 times.
[0041]
Allergens were detected using ECL + Plus, a Western blotting detection system manufactured by Amersham, according to the manufacturer's manual. Other operating procedures were performed according to general methods.
[0042]
A calibration curve related to the amount of mite allergen in the mite extract was obtained from the amount of luminescence obtained depending on the concentration of the mite extract. FIG. 4 is a schematic diagram showing the results of Western blotting. As is clear from FIG. 4, the amount of luminescence obtained increased in a concentration-dependent manner. Therefore, it became clear that the amount of protein in the mite extract reflects the amount of mite allergen. Further, FIG. 5 shows a calibration curve obtained by quantifying the amount of luminescence obtained by Western blotting.
[0043]
Example 2 Allergen Measurement 2
In the following examples, including this example, the relative amount of mite allergen in the sample was measured by tick scan. First, in order to confirm the quantitative property of tick scan, a calibration curve for mite allergen in mite extract was prepared.
[0044]
For the solution obtained by diluting the stock mite solution with a PBS solution, allergens were detected using a tick scan (DaniScan, registered trademark, manufactured by Asahi Breweries) as a mite allergen measurement system.
[0045]
The tick scan is a kit that can easily detect mite allergen using an antigen-antibody reaction. Band 3 reflecting the amount of mite allergen in the test sample when the test sample is added to the sample addition section 2 of the dust collector 1 provided in the mite scan shown in FIG. 6 and the developer is added thereto. Is obtained. It is possible to detect a relative allergen in the test sample from the band. In this specification, for convenience, each band of tick scan is indicated by “thick solid line”, “medium thick solid line”, “thin solid line”, “dashed line”, and “no display”. Four levels of color intensity are schematically shown in the order of “medium thick solid line”> “thin solid line”> “broken line”> “no display”.
[0046]
The amount of mite allergen contained in the sample is the peak value (T) of the color intensity derived from the mite antibody bound to the mite antigen in the mite extract and the color intensity derived from the mite antibody not bound to the mite antigen. From the peak value (C), the peak ratio was evaluated according to the following formula. FIG. 7 shows a graph showing the relationship between the position on the tick scan and the color intensity. Here, the peak value (T) and the peak value (C) correspond to the peaks of “C” and “T” in FIG.
[0047]
The formula for obtaining the peak ratio from the color intensity of peak C and peak T from the color intensity is:
Peak ratio (%) = T / (T + C) * 100
It is. At this time, if C> T, the amount of mite allergen in the sample is small, and if T> C, it indicates that a large amount of mite allergen is present in the sample.
[0048]
FIG. 6 is a schematic diagram of a tick scan showing a mite allergen positive (that is, when there are many allergens). The band at the position C is a band derived from the mite antibody that did not bind to the mite antigen, and the band at the position T is a band derived from the mite antibody bound to the mite antigen. FIG. 8 shows a calibration curve for mite allergens present in the mite extract.
[0049]
Example 3 Inactivation of allergens by heat
The following experiment was conducted to prove the mite allergen inactivation effect by heat treatment.
[0050]
A 10-fold diluted solution of tick stock solution in PBS was prepared. The obtained 10-fold diluted solution was dispensed into 7 microtubes, and one of them was used as a control group with a heating time of 0 minutes. The remaining six were heated to 80 ° C. on a heat block and maintained for 10, 20, 30, 40, 50 and 60 minutes, respectively. This was designated as a treatment zone. After the heating, the mixture was cooled on ice. Thereafter, 5 μL of each sample was extracted and added to a tick scan to detect allergen. Since the added 5 μL contains 2 μg of mite extract as a protein, 2 μg of mite extract is added per tick scan.
[0051]
The graph showing the relationship between the heat treatment time and the amount of mite allergen in the sample is shown in the graph of FIG. The vertical axis of the graph of FIG. 9 shows the relative peak ratio in the treatment group, where the peak ratio in the control group is 100, and the horizontal axis shows the heat treatment time.
[0052]
As is clear from the graph, the allergen was inactivated by heat treatment. In particular, considerable allergen was inactivated after 1 hour of heat treatment (FIG. 9).
[0053]
Example 4 Inactivation by alkali
The following experiment was conducted to prove the mite allergen inactivation effect by alkali treatment.
[0054]
First, 50 μL of mite stock solution and 50 μL of 5M sodium hydroxide were mixed in 5 microtubes on ice. The final concentration of sodium hydroxide in the resulting mixture is 2.5M. The microtubes were maintained under temperature conditions of 40 ° C. and 60 ° C. for 0, 10 and 30 minutes, respectively. After completion of the reaction, 50 μL of 5M hydrochloric acid was added to neutralize on ice. Thereafter, 1 μL of each of the obtained reaction solutions (1.33 μg mite extract / tick scan as a protein) was added to the tick scan, and mite allergen was detected. The results are shown in FIG. FIG. 10 is a diagram schematically showing a band seen in the tick scan after the reaction. 1 in FIG. 10 is the result obtained without treatment (ie, the result of adding only 1.33 μg mite extract per tick scan), which is a control sample. 2 to 4 in FIG. 10 are the results of alkali treatment of the mite extract at 40 ° C. for 0 minutes, 10 minutes, and 30 minutes, respectively. 5 and 6 in FIG. 10 are the results when the mite extract was alkali-treated at 60 ° C. for 0 minute and 10 minutes, respectively. As is clear from FIG. 10, allergens were inactivated by treatment for 10 minutes at both 40 ° C. and 60 ° C. (FIG. 10).
[0055]
Example 5 Inactivation by acid
The following experiment was conducted to prove the mite allergen inactivation effect by acid treatment.
[0056]
First, 50 μL of mite stock solution and 50 μL of 5M hydrochloric acid were mixed on four microtubes on ice. The final concentration of hydrochloric acid in the obtained mixed solution is 2.5M. The microtubes were maintained under a temperature condition of 60 ° C. for 0, 10, 30 and 60 minutes, respectively. After completion of the reaction, 50 μL of 5M sodium hydroxide was added to neutralize on ice. 1 μL of each of the obtained reaction solutions (0.133 μg mite extract / tick scan as protein) was added to the tick scan, and the amount of mite allergen was measured.
[0057]
The results are shown in FIG. FIG. 10 is a diagram schematically showing a band seen in the tick scan after the reaction. In FIG. 11, 1 was no treatment (that is, 1.33 μg of mite extract was added as a protein per tick scan), and this was used as a control sample. 2 to 5 in FIG. 11 show the results when the mite extract was acid-treated at 60 ° C. for 0 minutes, 10 minutes, 30 minutes and 60 minutes, respectively. As is clear from FIG. 11, it was confirmed that most allergens were inactivated by acid treatment at 60 ° C. for 60 minutes.
[0058]
Example 6 Inactivation by Protease 1
The following experiment was conducted to prove the mite allergen inactivation effect by protease.
[0059]
First, 90 μL of mite stock solution and 8 μL (0.8 units) of pfu protease S (hereinafter referred to as “pfu”, Takara Shuzo Co., Ltd.) were added to each of the three microtubes on ice. Of these, one was heated at 0 ° C. for 10 minutes, the other one was heated at 95 ° C. for 10 minutes, and the other one was heated at 80 ° C. for 10 minutes. The control sample without enzyme addition was heated at 95 ° C. for 10 minutes. After these heat treatments, all the microtubes were returned to ice, and a solution of phenylmethylsulfonyl fluoride (hereinafter referred to as PMSF) as a protease inhibitor was added (final concentration of PMSF is 4 mM).
[0060]
Thereafter, 5 μL was extracted from each sample to which PMSF was added and added to a tick scan to detect mite allergen. At this time, 18 μg of mite extract is included as the amount of protein per tick scan.
[0061]
The results are shown in FIG. 12 in FIG. 12 is a sample added with enzyme and reacted at 0 ° C. for 10 minutes, 2 in FIG. 12 is a control sample maintained at 95 ° C. for 10 minutes without enzyme, and 3 is a sample treated at 95 ° C. for 10 minutes with addition of enzyme. Indicates the results for the sample treated for 10 minutes at 80 ° C. with enzyme addition. As is apparent from the figure, mite allergen activity can be inactivated by protease treatment at the optimum temperature described above for 10 minutes.
[0062]
Example 7 Inactivation by Protease 2
Next, an experiment was conducted to prove the mite allergen inactivation effect when protease and urea were used in combination.
[0063]
To each of the three microtubes, 20 μL of mite stock solution and 3.2 μL (0.32 units) of pfu were added on ice, and 16.8 μL of 10 M urea prepared in PBS solution was further added. The final concentration of urea is 4.2M. They were then heated at 60 ° C. for 0, 10 and 30 minutes. After heating, the reaction was completed after returning to ice. 1 μL of each sample was extracted and allergen was detected by tick scanning. At this time, the tick extract added to one tick scan has a protein amount of 2 μg. The results are shown as 1 to 3 in FIG. 1, 2 and 3 are the results for samples heated at 60 ° C. for 0, 10 and 30 minutes, respectively.
[0064]
Further, the following tests were conducted in the same manner. To each of the four microtubes, 12 μL of mite stock solution and 8 μL (0.8 units) of pfu were added on ice, and 80 μL of 10 M urea was further added. The final concentration of urea is 8M. These were then heated at 40 ° C. for 0, 10, 30 and 60 minutes. 1 μL of each sample was extracted and the amount of allergen was measured by tick scanning. At this time, the mite extract added to one tick scan has a protein amount of 0.48 μg. The results are shown in 5 to 8 in FIG. 5, 6, 7 and 8 in FIG. 13 are the results for samples heated at 40 ° C. for 0, 10, 30 and 60 minutes, respectively.
[0065]
As a control sample, a sample without enzyme and heat treatment was prepared and tested. On ice, 12 μL of tick stock solution and 80 μL of 10 M urea and 8 μL of PBS solution were added to one microtube. The final concentration of urea is 8M. This microtube was not heated. 1 μL was extracted from this sample, and allergen was detected by tick scanning. At this time, the mite extract added to one tick scan has a protein amount of 0.48 μg. The result is shown as 4 in FIG.
[0066]
As is clear from the results shown in FIG. 13, the allergen was inactivated after 10 minutes when heated at 60 ° C. in the presence of 4.2 M urea. When heated at 40 ° C. in the presence of urea 8M, the allergen was inactivated after 30 minutes.
[0067]
Example 8 Inactivation by Protease 3
Furthermore, an experiment was conducted to prove mite allergen inactivation when papain was used as another protease example.
[0068]
First, 0.2 g / mL papain (manufactured by Nagase ChemteX Corporation, purified papain for food) and 100 μL of PBS solution were mixed with 400 μL of 10 M urea solution to prepare 8 M urea papain solution.
[0069]
20 μL of mite stock solution and 80 μL of 8M urea papain solution were added to each of the 8 microtubes. The final concentration of urea is 6.4M. These were heated at 40 ° C. for 0, 10, 30, 60, 120, 165, 280 and 320 minutes. After the heating, 18 μL each was taken out from each sample, and 2 μL each of 10 mM antipain (antipain, manufactured by BACHEM) was added. The final concentration of antipine is 1 mM.
[0070]
1 μL of each of the obtained solutions was collected and added to a tick scan, and mite allergen was detected. At this time, 0.72 μg of mite extract was added as a protein per tick scan.
[0071]
The obtained results are shown in 1 to 8 of FIG. 1, 2, 3, 4, 5, 6, 7, and 8 are the results for samples obtained at 0, 10, 30, 60, 120, 165, 280, and 320 minutes at 40 ° C., respectively. . From these results, it was possible to inactivate allergens by heating for 165 minutes under the above conditions, and most allergens were inactivated particularly by heating for 320 minutes.
[0072]
Furthermore, the effect of papain when heated at 30 ° C. was tested.
[0073]
20 μL of tick stock solution and 180 μL of 8M urea papain solution were added to each of the five microtubes. The final concentration of urea is 7.2M. These were heated at 30 ° C. for 0, 10, 30, 120 and 180 minutes. After the heating, 18 μL each was taken out from each sample, and 2 μL each of 10 mM antipain (antipain, manufactured by BACHEM) was added. The final concentration of antipine is 1 mM.
[0074]
1 μL of each of the obtained solutions was added to the tick scan, and mite allergen was detected. At this time, 0.36 μg of mite extract was added as a protein per tick scan.
[0075]
The obtained results are shown in 9 to 13 of FIG. 9, 10, 11, 12, and 13 in FIG. 14 are the results of the samples obtained by performing at 0, 10, 30, 120, and 180 minutes at 30 ° C., respectively. Under the above conditions, allergens could be inactivated by heating for 120 minutes (FIG. 14). Further, although not shown in FIG. 14, most allergens could be inactivated by heating for 320 minutes under the above conditions.
[0076]
As is clear from the above examples, according to the present invention, a universally usable solution is provided as a method for inactivating allergens in which a protein becomes an epitope. In addition, the present invention can deal with all allergens in which proteins are epitopes.
[0077]
(2) Examples of filters and air conditioning
Embodiments of a filter and an air conditioner according to the present invention will be described below with reference to the drawings.
[0078]
Example 9
Example 9 will be described with reference to FIG. The number 11 in the figure indicates a non-woven filter that traps allergens. On the lower surface of the nonwoven fabric filter 11, a stainless steel fibrous surface heating element 12 having a mesh smaller than the diameter of pollen particles (20 to 30 μm) and mites (particularly mite feces, 10 to 40 μm) is disposed. . An electric heater 13 for heating the surface heating element 12 is disposed below the stainless steel fibrous surface heating element (trade name: Softeleck, manufactured by Teijin Ltd.) 12.
[0079]
Thus, the allergen inert filter 14 according to Example 9 is a stainless steel surface having a non-woven fabric filter 11 for trapping allergens and a mesh smaller than the diameter of pollen particles and ticks arranged on the lower surface of the non-woven fabric filter 11. The heating element 12 and the electric heater 13 that heats the surface heating element 12 disposed below the fibrous surface heating element 12 are provided.
[0080]
Such an allergen inert filter 14 is attached to a predetermined position of an air conditioner (see FIG. 15) described later with a size (for example, 5 cm × 10 cm) that can be mounted on an air conditioner. When the allergen inert filter 14 is attached to the air intake port of the air conditioner, the nonwoven fabric filter 11 is disposed on the inlet side of the allergen-containing air (arrow A). When the air conditioner is OFF, the electric heater 13 of the filter 14 is turned ON and the surface heating element 12 is heated at a heating temperature of about 70 ° C., for example.
[0081]
As a result, the allergen-containing air passes through the nonwoven fabric filter 11, but pollen particles and ticks are captured by the surface heating element 12 held at a high temperature, where the allergen protein is denatured and the activity as an allergen is lost. Allergen-reducing air (arrow B) passes from the stainless steel surface heating element 12.
[0082]
Therefore, according to the allergen inactive filter 14 of Example 1, since no chemical for pest control is used as in the prior art, the influence of the chemical itself on the human body can be avoided, and the heating element is kept at a predetermined temperature. The amount of allergen can be automatically reduced by simply heating with
[0083]
Example 10
Please refer to FIG. The allergen inert filter 21 according to Example 10 has a configuration in which a non-woven fabric filter 22 is held with a strongly acidic cation exchange resin (not shown). Here, the strongly acidic cation exchange resin is R—SO 3 .H (where R represents a polymer substrate), and can be regenerated using an acid. This strongly acidic cation exchange resin is activated before use.
[0084]
According to Example 10, the allergen protein is denatured by the strong acid cation exchange resin held by the non-woven fabric filter 22 just by passing the allergen-containing air A through the non-woven fabric filter 22 and its activity as an allergen Lose. Therefore, allergen-reduced air B is exhausted from the nonwoven fabric filter 22. Therefore, according to the allergen inert filter 21 of Example 10, since no chemical for pest control is used as in the prior art, the influence of the chemical itself on the human body can be avoided and the strongly acidic cation exchange resin is retained. The allergen amount can be automatically reduced simply by passing the allergen-containing air A through the non-woven fabric filter 22 that has been made.
[0085]
In addition, in Example 10, although the case where strong acidic cation exchange resin was used was described, not only this but the strong basic anion exchange resin shown to the said compound 1 may be used. However, also in this case, it is preferable to activate the strongly basic anion exchange resin before use.
[0086]
Example 11
Embodiment 11 will be described with reference to FIG. Reference numeral 31 in the figure indicates a first support made of resin having a size that the water-absorbing polymer 32 does not move and a mesh (> 50 μm) through which pollen particles and ticks are passed. A resin-made second support 33 having a mesh size (> 50 μm) through which the water-absorbing polymer 34 does not move and through which pollen particles and mites do not pass is disposed below the first support 31. ing. Between the first support 31 and the second support 33, a water-absorbing polymer layer 34 made of a water-absorbing polymer 32 is sandwiched. Here, the water-absorbing polymer 32 contains, for example, a protease of 500,000 units / filter. The water-absorbing polymer layer 34 can be formed, for example, by impregnating a polymer with an oxygen solution containing protease.
[0087]
The allergen inert filter 35 according to Example 11 has a structure in which a water-absorbing polymer layer 34 is sandwiched between a first support 31 and a second support 33 made of resin. Thus, the allergen-containing air A only passes through the water-absorbing polymer layer 34, and the allergen is decomposed by the enzyme that decomposes the protein. Therefore, according to the allergen inert filter 35 of Example 11, since no chemical for pest control is used as in the prior art, the influence of the chemical itself on the human body can be avoided, and the water-absorbing polymer layer 34 contains allergen. The amount of allergen can be automatically reduced simply by passing air A.
[0088]
Example 12
FIG. 15 shows an example in which the allergen inert filter according to Examples 9 to 11 is attached to an air intake port of an air conditioner. In FIG. 15, reference numeral 42 indicates a cooling unit for an air conditioner, and reference numeral 43 indicates a housing. Inside the housing 43, a fan 44 and the like are disposed. Here, the allergen inert filter 14 is arranged so that the nonwoven fabric filter 21 which is the main component of the filter 14 is located at the inlet of the air allergen-containing air.
[0089]
Thus, the air conditioner according to Example 12 has a configuration in which the allergen inert filter 14 is disposed at the air intake port of the air conditioner. Therefore, when the air conditioner is off, the electric heater 13 of the filter 14 is turned on to cause the surface heating element 12 to generate heat, whereby pollen particles and ticks are captured by the surface heating element 12 held at a high temperature. Allergen protein is denatured and allergen activity is lost. Allergen-reduced air passes from the stainless steel surface heating element 12. The electric heater 13 uses an air conditioner power source.
[0090]
The living environment can be significantly improved by adding the allergen decomposition function of the present invention to an air conditioner such as an air conditioner or an air purifier.
[0091]
【The invention's effect】
In accordance with the present invention, a method for specifically eliminating allergens is provided.
[Brief description of the drawings]
FIG. 1 shows an example of an allergen inactive filter according to the present invention.
FIG. 2 is a diagram showing an example of an allergen inactive filter according to the present invention.
FIG. 3 shows an example of an allergen inactive filter according to the present invention.
FIG. 4 is a schematic diagram of the results of Western blotting.
FIG. 5 is a graph showing a calibration curve obtained by Western blotting.
FIG. 6 is a schematic diagram of a tick scan showing a band obtained when a large amount of mite allergen is present in a sample.
FIG. 7 is a graph showing the relationship between the position of a band on a tick scan and the color development intensity.
FIG. 8 is a graph showing a calibration curve for mite allergens present in mite extracts.
FIG. 9 is a graph showing the relationship between heat treatment time and the amount of mite allergen in a sample.
FIG. 10 is a diagram schematically showing a band seen in a tick scan after reaction.
FIG. 11 is a diagram schematically showing a band seen in a tick scan after reaction.
FIG. 12 is a diagram schematically showing a band seen in a tick scan after reaction.
FIG. 13 is a diagram schematically showing a band seen in a tick scan after reaction.
FIG. 14 is a diagram schematically showing a band seen in a tick scan after reaction.
15 is a view showing an air conditioner according to the present invention in which the allergen inert filter of FIG. 1 is attached to the air inlet side.
[Explanation of symbols]
1. Dust collector 1. 2. Sample addition section Band 11. Non-woven fabric filter 12. Fibrous surface heating element 13. Electric heater 14. Allergen inactive filter 21,22. Nonwoven fabric filter 31. First support 32. Water-absorbing polymer 33. Second support 34. Water-absorbing polymer layer 35. Allergen inert filter 42. Air conditioning cooling unit 43. Housing 44. fan

Claims (3)

An allergen inactivation method comprising inactivating the allergen by maintaining the allergen in the presence of protease and urea. A Reruge emissions inactivation how according to claim 1, characterized in that also present more heat. An air conditioner comprising an allergen inactive part for performing the method for inactivating allergens according to claim 1 or 2 .

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