JPS6116940B2 - - Google Patents
Info
- Publication number
- JPS6116940B2 JPS6116940B2 JP3316980A JP3316980A JPS6116940B2 JP S6116940 B2 JPS6116940 B2 JP S6116940B2 JP 3316980 A JP3316980 A JP 3316980A JP 3316980 A JP3316980 A JP 3316980A JP S6116940 B2 JPS6116940 B2 JP S6116940B2
- Authority
- JP
- Japan
- Prior art keywords
- latex
- antibody
- antibodies
- sensitized
- reagent
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired
Links
- 239000004816 latex Substances 0.000 claims description 56
- 229920000126 latex Polymers 0.000 claims description 56
- 239000003153 chemical reaction reagent Substances 0.000 claims description 25
- 239000002245 particle Substances 0.000 claims description 17
- 210000002966 serum Anatomy 0.000 claims description 14
- 230000001235 sensitizing effect Effects 0.000 claims description 5
- 239000004793 Polystyrene Substances 0.000 claims description 4
- 239000007788 liquid Substances 0.000 claims description 4
- 229920002223 polystyrene Polymers 0.000 claims description 4
- 238000004519 manufacturing process Methods 0.000 claims description 3
- 239000000427 antigen Substances 0.000 description 17
- 102000036639 antigens Human genes 0.000 description 16
- 108091007433 antigens Proteins 0.000 description 16
- 238000006243 chemical reaction Methods 0.000 description 14
- 230000004520 agglutination Effects 0.000 description 12
- LOKCTEFSRHRXRJ-UHFFFAOYSA-I dipotassium trisodium dihydrogen phosphate hydrogen phosphate dichloride Chemical compound P(=O)(O)(O)[O-].[K+].P(=O)(O)([O-])[O-].[Na+].[Na+].[Cl-].[K+].[Cl-].[Na+] LOKCTEFSRHRXRJ-UHFFFAOYSA-I 0.000 description 10
- 239000002953 phosphate buffered saline Substances 0.000 description 10
- 238000000034 method Methods 0.000 description 7
- 102000004169 proteins and genes Human genes 0.000 description 6
- 108090000623 proteins and genes Proteins 0.000 description 6
- 206010070834 Sensitisation Diseases 0.000 description 4
- 239000000872 buffer Substances 0.000 description 4
- 230000000052 comparative effect Effects 0.000 description 4
- 230000008313 sensitization Effects 0.000 description 4
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 3
- 241000700199 Cavia porcellus Species 0.000 description 3
- 238000004220 aggregation Methods 0.000 description 3
- 230000002776 aggregation Effects 0.000 description 3
- 229940098773 bovine serum albumin Drugs 0.000 description 3
- 230000035945 sensitivity Effects 0.000 description 3
- 239000007787 solid Substances 0.000 description 3
- PPBRXRYQALVLMV-UHFFFAOYSA-N Styrene Chemical compound C=CC1=CC=CC=C1 PPBRXRYQALVLMV-UHFFFAOYSA-N 0.000 description 2
- 239000007853 buffer solution Substances 0.000 description 2
- 238000005119 centrifugation Methods 0.000 description 2
- 238000012017 passive hemagglutination assay Methods 0.000 description 2
- 102100023635 Alpha-fetoprotein Human genes 0.000 description 1
- 241000700198 Cavia Species 0.000 description 1
- 238000011891 EIA kit Methods 0.000 description 1
- 241000701076 Macacine alphaherpesvirus 1 Species 0.000 description 1
- 241000283977 Oryctolagus Species 0.000 description 1
- 238000001042 affinity chromatography Methods 0.000 description 1
- 108010026331 alpha-Fetoproteins Proteins 0.000 description 1
- 239000000470 constituent Substances 0.000 description 1
- 229920001577 copolymer Polymers 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 208000002672 hepatitis B Diseases 0.000 description 1
- 229920001519 homopolymer Polymers 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 230000035935 pregnancy Effects 0.000 description 1
- 239000000243 solution Substances 0.000 description 1
- 229920003048 styrene butadiene rubber Polymers 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 229920003002 synthetic resin Polymers 0.000 description 1
- 239000000057 synthetic resin Substances 0.000 description 1
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
- G01N33/54313—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals the carrier being characterised by its particulate form
Landscapes
- Health & Medical Sciences (AREA)
- Immunology (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Molecular Biology (AREA)
- Biomedical Technology (AREA)
- Chemical & Material Sciences (AREA)
- Hematology (AREA)
- Urology & Nephrology (AREA)
- Biotechnology (AREA)
- Microbiology (AREA)
- Cell Biology (AREA)
- Food Science & Technology (AREA)
- Medicinal Chemistry (AREA)
- Physics & Mathematics (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Pathology (AREA)
- Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
Description
本発明は診断用ラテツクス試薬に関する。
粒径0.01〜１μのラテツクス粒子に抗原又は抗
体を感作させ、弱アルカリ性の緩衝液に分散させ
た診断用ラテツクス試薬は、体液中のそれぞれ対
応する抗体又は抗原と特異的に反応し、可視的な
凝集反応を起こすので、現在、リウマチ因子の検
出や妊娠診断等に広く用いられている。
このようなラテツクス試薬は、一般に測定対象
である抗原又は抗体に対応する抗体又は抗原をラ
テツクス粒子表面に吸着させることにより得られ
るが、このように単に抗体又は抗原を吸着させる
だけでは、体液中には本来、多様なタンパク質が
含まれているため、目的とする抗原又は抗体以外
のものがラテツクス試薬と反応し、非特異的凝集
反応を起こすことがある。
ラテツクス試薬におけるこの非特異的凝集反応
を防ぐために、抗体又は抗原を担体粒子に吸着さ
せる前に、目的とする抗原抗体反応に関与しない
不活性なタンパク質で担体粒子を被覆することが
提案されているが（特公昭43−12741号）、しか
し、この方法によれば、抗体又は抗原が担体粒子
に吸着し難い欠点がある。このため、抗体又は抗
原を担体粒子に感作させた後、目的とする抗体抗
原反応に関与しない不活性なタンパク質で被覆す
る方法も提案されているが（特公昭49−11407
号）、この場合には、得られるラテツクス試薬の
感度が小さい。
本発明は上記の問題を解決するためになされた
ものであり、非特異的凝集反応を阻止すると共
に、感度が非常に高い診断用ラテツクス試薬を提
供することを目的とする。
本発明の診断用ラテツクス試薬の製造方法は、
ラテツクス粒子に抗体を感作させ、次いでこの抗
体感体ラテツクスを、上記と同じ抗体を含む血清
を含有する液中に分散させたのち該液から分離す
ることにより処理することを特徴とするものであ
る。
ラテツクスとしては種々の合成樹脂ラテツクス
が用いられるが、好ましくはスチレンを構成単位
とする単独重合体及び共重合体が用いられ、具体
例としてポリスチレン、スチレン−ブタジエン共
重合体等のラテツクスを挙げることができる。ラ
テツクス粒子の大きさは0.05〜１μが適当であ
る。ラテツクスは通常、弱アルカリ性のリン酸緩
衝液（PBS）のような緩衝液に加えて分散させ、
抗体を溶解した緩衝液に上記ラテツクスを加えて
混合し、抗体を感作させる。感作温度は25〜45
℃、好ましくは30〜37℃であり、感作時間は数分
乃至数時間である。普通、１時間以内で十分であ
る。次に、このようにして抗体を感作したラテツ
クスを遠心分離して、未吸着の抗体を除く。この
ようなラテツクスへの抗体の感作方法は既によく
知られており、本発明においては、上に例として
挙げた方法に何ら限定されるものではない。
このようにして得た抗体感作ラテツクスを本発
明に従つて処理するための抗体を含む血清、即ち
抗血清は、通常、この抗血清を0.1〜20重量％濃
度で含有し、従つて、抗体を１〜1000μｇ／c.c.程
度の濃度で含有する緩衝液として用いられる。こ
のような緩衝液に上で得た抗体感作ラテツクスを
懸濁させて抗血清処理するが、この処理温度及び
処理時間は前記したラテツクスの抗体感作の場合
と同様である。
この抗血清処理後、遠心分離して得たラテツク
ス粒子を再び緩衝液に分散して、本発明のラテツ
クス試薬を得る。ラテツクスにおける固型分は、
従来の試薬と同様に、0.5〜３重量％でよい。所
望ならば、感作ラテツクスに直接抗血清を加えて
もよい。
本発明の方法は、ラテツクス粒子に抗体を感作
した後、目的とする抗原抗体反応に関与しない不
活性なタンパク質で処理する従来知られている方
法と異なり、以上のように、感作した抗体を含む
血清にて処理するものであり、これによつてラテ
ツクス試薬の非特異的凝集反応をよく阻止すると
同時に、以下の実施例に示すように、測定活性を
飛躍的に高めることができたのである。
以下に本発明の実施例を挙げる。尚、以下にお
いて、凝集の強さは次のように表示した。
〓：30秒以那内に強く凝集する。
〓：２分以内に強く凝集する。
＋：５分以内に凝集する。
−：30分以後も凝集しない。
実施例 １
平均粒径0.47μのポリスチレンラテツクスを固
型分２％となるようにPH7.4のPBSに分散させ
た。別に、モルモツトの産出した抗Ｂ型肝炎ウイ
ルス表面抗原（HBs抗原）モノスペシフイツク抗
体を上記と同様のPBSに40μｇ／c.c.の濃度に溶解
し、この溶液１容と前記ラテツクス１容とを混合
した後、37℃の温度で２時間インキユベートし、
ラテツクス粒子に抗体を感作させた。この感作ラ
テツクスを15分間、15000rpmで遠心分離し、未
吸着の抗体を除去して、感作ラテツクスを分離し
た。尚、上清中の抗体価は受身赤血球凝集反応法
（PHA法）にて測定したところ、少なくとも99.5
％の抗体がラテツクス粒子に吸着されたことが判
つた。
次に、HBs抗原で免疫されたモルモツトの抗血
清をアフイニテイクロマトグラフにより処理し
て、抗人タンパク抗体を除き、こうして得たモル
モツトの抗血清を５重量％濃度になるように、
PBSを加えた。この抗血清を含むPBS １容（こ
のPBSには抗体HBs抗体が約50μｇ／c.c.含まれて
いる。）に上で得た抗体感作ラテツクス粒子を分
散させ、37℃で10分間インキユベートした。この
ようにして抗血清処理した感作ラテツクスを
12000rpmで遠心分離し、沈降したラテツクス粒
子をPH７のPBSに再分散させ、本発明によるラテ
ツクス試薬（固型分２重量％）を得た。
このラテツクス試薬を用い、種々の濃度でHBs
抗原を含むヒト血清に対する凝集の強さを測定し
て、第１表の結果を得た。
The present invention relates to diagnostic latex reagents. Diagnostic latex reagents are made by sensitizing latex particles with a particle size of 0.01 to 1 μm with antigens or antibodies and dispersing them in a weakly alkaline buffer solution. Because it causes an agglutination reaction, it is currently widely used for detecting rheumatoid factors and diagnosing pregnancy. Such latex reagents are generally obtained by adsorbing antibodies or antigens corresponding to the antigens or antibodies to be measured onto the surface of latex particles. Since it originally contains a variety of proteins, substances other than the target antigen or antibody may react with the latex reagent, causing a nonspecific agglutination reaction. In order to prevent this non-specific agglutination reaction in latex reagents, it has been proposed to coat the carrier particles with an inactive protein that does not participate in the desired antigen-antibody reaction before adsorbing antibodies or antigens to the carrier particles. (Japanese Patent Publication No. 43-12741) However, this method has the disadvantage that antibodies or antigens are difficult to adsorb to carrier particles. For this reason, a method has also been proposed in which carrier particles are sensitized with antibodies or antigens and then coated with an inactive protein that does not participate in the desired antibody-antigen reaction (Japanese Patent Publication No. 11407-1971).
In this case, the sensitivity of the latex reagent obtained is low. The present invention was made to solve the above problems, and aims to provide a diagnostic latex reagent that prevents nonspecific agglutination reactions and has extremely high sensitivity. The method for producing a diagnostic latex reagent of the present invention includes:
It is characterized by sensitizing latex particles with antibodies, then dispersing the antibody-sensing latex in a liquid containing serum containing the same antibodies as above, and then separating it from the liquid. be. Various synthetic resin latexes can be used as the latex, but homopolymers and copolymers having styrene as a constituent unit are preferably used, and specific examples include latexes such as polystyrene and styrene-butadiene copolymers. can. The appropriate size of the latex particles is 0.05 to 1 micron. Latexes are usually dispersed in a buffer such as slightly alkaline phosphate buffered saline (PBS).
The latex is added to a buffer in which the antibody has been dissolved and mixed to sensitize the antibody. Sensitization temperature is 25-45
℃, preferably 30 to 37℃, and the sensitization time is several minutes to several hours. Usually less than 1 hour is sufficient. Next, the latex sensitized with antibodies in this manner is centrifuged to remove unadsorbed antibodies. Such methods of sensitizing antibodies to latex are already well known, and the present invention is not limited to the methods listed above as examples. The antibody-containing serum, ie, the antiserum, for treating the antibody-sensitized latex obtained in this way according to the present invention usually contains this antiserum at a concentration of 0.1 to 20% by weight, and therefore contains the antibody. It is used as a buffer containing a concentration of about 1 to 1000 μg/cc. The antibody-sensitized latex obtained above is suspended in such a buffer and treated with antiserum, and the treatment temperature and treatment time are the same as in the case of antibody sensitization of the latex described above. After this antiserum treatment, the latex particles obtained by centrifugation are again dispersed in a buffer solution to obtain the latex reagent of the present invention. The solid content in latex is
As with conventional reagents, it may be 0.5-3% by weight. If desired, antiserum may be added directly to the sensitized latex. The method of the present invention differs from conventionally known methods in which latex particles are sensitized with antibodies and then treated with an inactive protein that does not participate in the target antigen-antibody reaction. This treatment effectively inhibits the nonspecific agglutination reaction of the latex reagent, and at the same time dramatically increases the measurement activity, as shown in the examples below. be. Examples of the present invention are listed below. In addition, below, the strength of aggregation is expressed as follows. 〓: Strongly aggregates within 30 seconds. 〓: Strongly aggregates within 2 minutes. +: Aggregation occurs within 5 minutes. -: No aggregation after 30 minutes. Example 1 Polystyrene latex having an average particle size of 0.47 μm was dispersed in PBS having a pH of 7.4 to a solid content of 2%. Separately, an anti-hepatitis B virus surface antigen (HBs antigen) monospecific antibody produced by guinea pigs was dissolved in the same PBS as above to a concentration of 40 μg/cc, and 1 volume of this solution was mixed with 1 volume of the latex. After that, incubate at a temperature of 37℃ for 2 hours,
Latex particles were sensitized with antibodies. This sensitized latex was centrifuged at 15,000 rpm for 15 minutes to remove unadsorbed antibodies and separate the sensitized latex. The antibody titer in the supernatant was determined to be at least 99.5 when measured by passive hemagglutination assay (PHA method).
It was found that % of the antibody was adsorbed to the latex particles. Next, the guinea pig antiserum immunized with HBs antigen was treated with affinity chromatography to remove anti-human protein antibodies, and the guinea pig antiserum thus obtained was adjusted to a concentration of 5% by weight.
PBS was added. The antibody-sensitized latex particles obtained above were dispersed in 1 volume of PBS containing this antiserum (this PBS contains about 50 μg/cc of HBs antibody) and incubated at 37° C. for 10 minutes. The sensitized latex treated with antiserum in this way is
After centrifugation at 12,000 rpm, the sedimented latex particles were redispersed in PBS at pH 7 to obtain a latex reagent (solid content: 2% by weight) according to the present invention. Using this latex reagent, HBs was added at various concentrations.
The strength of agglutination against human serum containing antigen was measured and the results shown in Table 1 were obtained.
【表】
次に、HBs抗原が４×10-10ｇ／c.c.以下である
1000人の正常人の血清について、本発明のラテツ
クス試薬を適用し、（HBs抗原検出用EIAキツト
「リバーセイア」（山之内製薬(株)）を用いて確
認）、上記と同様に凝集試験をしたところ、偽陽
性は僅かに１件であつた。また、HBs抗体を含有
する100人の血清については、偽陽性は１件もな
かつた。
比較例 １
感作ラテツクスを正常モルモツト血清で処理し
た以外は、実施例１と全く同様にして、抗HBs抗
体感作ラテツクスを得、実施例１と同様の凝集試
験をしたところ、第２表の結果を得た。[Table] Next, HBs antigen is 4×10 -10 g/cc or less.
When the latex reagent of the present invention was applied to the serum of 1000 normal people (confirmed using the EIA kit for detecting HBs antigen "Reversere" (Yamanouchi Pharmaceutical Co., Ltd.)), an agglutination test was conducted in the same manner as above. There was only one false positive. Furthermore, there were no false positives among sera from 100 people containing HBs antibodies. Comparative Example 1 An anti-HBs antibody sensitized latex was obtained in exactly the same manner as in Example 1, except that the sensitized latex was treated with normal guinea pig serum, and an agglutination test was conducted in the same manner as in Example 1. Got the results.
【表】
次に、このラテツクス試薬を実施例１と同様に
して1000検体の正常人血清に適用したところ、偽
陽性は８件であつた。また、抗体を有する血清と
の反応においては、偽陽性は３件であつた。
実施例 ２
実施例１と同じポリスチレンラテツクスに、実
施例１と同様にして、家兎の産出したα−フエト
プロテイン（α−FP）のモノスペシフイツク抗
体を感作した後、家兎の抗血清１重量％を含む
PBSで処理して、本発明のラテツクス試薬を得
た。このラテツクス試薬を用いて、ヒト血清中の
α−FPとの凝集反応を調べた。結果を第３表に
示す。[Table] Next, when this latex reagent was applied to 1000 normal human serum samples in the same manner as in Example 1, there were 8 false positives. Furthermore, in the reaction with serum containing antibodies, there were 3 false positives. Example 2 The same polystyrene latex as in Example 1 was sensitized with a monospecific antibody for α-fetoprotein (α-FP) produced by domestic rabbits in the same manner as in Example 1. Contains 1% by weight of antiserum of
The latex reagent of the present invention was obtained by treatment with PBS. Using this latex reagent, the agglutination reaction with α-FP in human serum was investigated. The results are shown in Table 3.
【表】
また、正常人の血清1000検体についての偽陽性
は２件であつた。
比較例 ２
実施例２で得たα−FP抗体感作ラテツクスを
牛血清アルブミン（BSA）１重量％を含有する
PBSを処理してラテツクス試薬を得た。この試薬
とヒト血清中のα−FPとの凝集反応を調べて、
第４表の結果を得た。[Table] Additionally, there were 2 false positives among 1000 normal human serum samples. Comparative Example 2 The α-FP antibody sensitized latex obtained in Example 2 containing 1% by weight of bovine serum albumin (BSA)
A latex reagent was obtained by treating PBS. We investigated the agglutination reaction between this reagent and α-FP in human serum.
The results shown in Table 4 were obtained.
【表】
また、このラテツクス試薬を正常人の血清1000
検体に適用したところ、偽陽性は121件にも達し
た。
以上から明らかなように、抗α−FP抗体を感
作した後、目的とする抗原抗体反応に関与しない
不活性なタンパク質BSAで処理する従来方法の
ラテツクス試薬によれば（比較例２）、非特異的
凝集反応が著しく、且つ、ヒト血清中の10-5ｇ／
c.c.程度のα−FPしか検出し得ないが、本発明に
よつて、抗体感作後、抗血清にて処理したラテツ
クス試薬によれば（実施例２）、非特異的凝集反
応が殆んどなく、且つ、ヒト血清中の10-8ｇ／c.c.
程度のα−FPを検出し得、従来品に比較して約
1000倍も高い感度を示す。
更に、HBs抗原検出用のラテツクス試薬の場
合、HBs抗体を感作した後、単に正常な血清で処
理するだけであれば（比較例１）、ヒト血清中の
10-5ｇ／c.c.程度の濃度のHBs抗原を検出するにと
どまるが、本発明に従つて抗血清で処理すれば
（実施例１）、10-8ｇ／c.c.程度のHBs抗原をも検出
することができる。[Table] In addition, this latex reagent was added to normal human serum 1000
When applied to samples, there were 121 false positives. As is clear from the above, according to the conventional latex reagent in which the anti-α-FP antibody is sensitized and then treated with BSA, an inactive protein that does not participate in the target antigen-antibody reaction (Comparative Example 2), The specific agglutination reaction is remarkable, and the concentration of 10 -5 g/
Although only α-FP of about cc can be detected, according to the present invention, with the latex reagent treated with antiserum after antibody sensitization (Example 2), there is almost no non-specific agglutination reaction. 10 -8 g/cc in human serum
It is possible to detect α-FP of approximately
Shows 1000 times higher sensitivity. Furthermore, in the case of a latex reagent for detecting HBs antigen, if it is simply treated with normal serum after sensitizing HBs antibody (Comparative Example 1),
HBs antigen at a concentration of about 10 -5 g/cc is only detected, but if treated with antiserum according to the present invention (Example 1), HBs antigen at a concentration of about 10 -8 g/cc can also be detected. be able to.
Claims (1)
の抗体感作ラテツクスを、上記と同じ抗体を含む
血清を含有する液中に分散させたのち該液から分
離することにより処理することを特徴する診断用
ラテツクス試薬の製造方法。 ２ ラテツクスがポリスチレン系ラテツクスであ
ることを特徴とする特許請求の範囲第１項記載の
診断用ラテツクス試薬の製造方法。[Claims] 1. Processing by sensitizing latex particles with antibodies, then dispersing the antibody-sensitized latex in a liquid containing serum containing the same antibodies as above, and separating it from the liquid. A method for producing a diagnostic latex reagent, characterized in that: 2. The method for producing a diagnostic latex reagent according to claim 1, wherein the latex is a polystyrene latex.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP3316980A JPS56129862A (en) | 1980-03-14 | 1980-03-14 | Production of diagnosis latex reagent |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP3316980A JPS56129862A (en) | 1980-03-14 | 1980-03-14 | Production of diagnosis latex reagent |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| JPS56129862A JPS56129862A (en) | 1981-10-12 |
| JPS6116940B2 true JPS6116940B2 (en) | 1986-05-02 |
Family
ID=12379027
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP3316980A Granted JPS56129862A (en) | 1980-03-14 | 1980-03-14 | Production of diagnosis latex reagent |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPS56129862A (en) |
Families Citing this family (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5258146A (en) * | 1988-09-26 | 1993-11-02 | 3D Systems, Inc. | Method of and apparatus for measuring and controlling fluid level in stereolithography |
| JPH0829426A (en) * | 1994-03-24 | 1996-02-02 | Yakult Honsha Co Ltd | Antibody sensitizing latex for detecting nitrate-forming bacteria and nitrite-forming bacterial |
| JPH095328A (en) * | 1995-06-19 | 1997-01-10 | Yakult Honsha Co Ltd | Antibody sensitized latex for detecting denitrifying bacteria and its detection method |
| JP4857813B2 (en) * | 2005-03-11 | 2012-01-18 | 東レ株式会社 | Coating apparatus, coating method, and method for producing coating film forming web |
| US8733275B2 (en) | 2005-03-11 | 2014-05-27 | Toray Industries, Inc. | Application apparatus, application method and method for manufacturing web having coating film |
-
1980
- 1980-03-14 JP JP3316980A patent/JPS56129862A/en active Granted
Also Published As
| Publication number | Publication date |
|---|---|
| JPS56129862A (en) | 1981-10-12 |
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