JPH03280881A - Method for immobilizing microorganism - Google Patents
Method for immobilizing microorganismInfo
- Publication number
- JPH03280881A JPH03280881A JP8109490A JP8109490A JPH03280881A JP H03280881 A JPH03280881 A JP H03280881A JP 8109490 A JP8109490 A JP 8109490A JP 8109490 A JP8109490 A JP 8109490A JP H03280881 A JPH03280881 A JP H03280881A
- Authority
- JP
- Japan
- Prior art keywords
- solution
- microorganisms
- contact
- polymer
- free amino
- 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.)
- Pending
Links
- 244000005700 microbiome Species 0.000 title claims abstract description 29
- 230000003100 immobilizing effect Effects 0.000 title claims abstract description 9
- 238000000034 method Methods 0.000 title claims description 12
- 238000006243 chemical reaction Methods 0.000 claims abstract description 17
- 239000003431 cross linking reagent Substances 0.000 claims abstract description 13
- 229920000642 polymer Polymers 0.000 claims abstract description 13
- 125000003277 amino group Chemical group 0.000 claims abstract description 11
- 125000000524 functional group Chemical group 0.000 claims abstract description 6
- 229920000729 poly(L-lysine) polymer Polymers 0.000 claims abstract description 6
- 238000005406 washing Methods 0.000 claims description 4
- SXRSQZLOMIGNAQ-UHFFFAOYSA-N Glutaraldehyde Chemical compound O=CCCCC=O SXRSQZLOMIGNAQ-UHFFFAOYSA-N 0.000 abstract description 5
- 230000035945 sensitivity Effects 0.000 abstract description 5
- 238000004132 cross linking Methods 0.000 abstract description 2
- 241000194017 Streptococcus Species 0.000 abstract 1
- 239000000243 solution Substances 0.000 description 15
- 241000283973 Oryctolagus cuniculus Species 0.000 description 10
- 102000004190 Enzymes Human genes 0.000 description 9
- 108090000790 Enzymes Proteins 0.000 description 9
- 238000003018 immunoassay Methods 0.000 description 9
- 230000004520 agglutination Effects 0.000 description 8
- 238000005259 measurement Methods 0.000 description 7
- 210000004027 cell Anatomy 0.000 description 6
- 241000222122 Candida albicans Species 0.000 description 5
- 230000001580 bacterial effect Effects 0.000 description 5
- 229940095731 candida albicans Drugs 0.000 description 5
- 241001505901 Streptococcus sp. 'group A' Species 0.000 description 4
- 239000000725 suspension Substances 0.000 description 4
- 241000894006 Bacteria Species 0.000 description 3
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 3
- 239000000427 antigen Substances 0.000 description 3
- 102000036639 antigens Human genes 0.000 description 3
- 108091007433 antigens Proteins 0.000 description 3
- KRKNYBCHXYNGOX-UHFFFAOYSA-N citric acid Chemical compound OC(=O)CC(O)(C(O)=O)CC(O)=O KRKNYBCHXYNGOX-UHFFFAOYSA-N 0.000 description 3
- 230000000694 effects Effects 0.000 description 3
- 239000008363 phosphate buffer Substances 0.000 description 3
- 241000233866 Fungi Species 0.000 description 2
- 239000004472 Lysine Substances 0.000 description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 2
- 239000011324 bead Substances 0.000 description 2
- 238000010586 diagram Methods 0.000 description 2
- 239000004033 plastic Substances 0.000 description 2
- 238000002360 preparation method Methods 0.000 description 2
- PUZPDOWCWNUUKD-UHFFFAOYSA-M sodium fluoride Chemical compound [F-].[Na+] PUZPDOWCWNUUKD-UHFFFAOYSA-M 0.000 description 2
- 239000007790 solid phase Substances 0.000 description 2
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 1
- 241001494479 Pecora Species 0.000 description 1
- 229920001213 Polysorbate 20 Polymers 0.000 description 1
- 239000004793 Polystyrene Substances 0.000 description 1
- 108010071390 Serum Albumin Proteins 0.000 description 1
- 102000007562 Serum Albumin Human genes 0.000 description 1
- 238000002835 absorbance Methods 0.000 description 1
- 125000003172 aldehyde group Chemical group 0.000 description 1
- 239000000783 alginic acid Substances 0.000 description 1
- 235000010443 alginic acid Nutrition 0.000 description 1
- 229920000615 alginic acid Polymers 0.000 description 1
- 229960001126 alginic acid Drugs 0.000 description 1
- 150000004781 alginic acids Chemical class 0.000 description 1
- 229940098773 bovine serum albumin Drugs 0.000 description 1
- 239000007853 buffer solution Substances 0.000 description 1
- 210000002421 cell wall Anatomy 0.000 description 1
- 239000003610 charcoal Substances 0.000 description 1
- 125000005442 diisocyanate group Chemical group 0.000 description 1
- 239000011521 glass Substances 0.000 description 1
- 239000002991 molded plastic Substances 0.000 description 1
- 230000003287 optical effect Effects 0.000 description 1
- 239000002245 particle Substances 0.000 description 1
- 102000013415 peroxidase activity proteins Human genes 0.000 description 1
- 108040007629 peroxidase activity proteins Proteins 0.000 description 1
- 239000000256 polyoxyethylene sorbitan monolaurate Substances 0.000 description 1
- 235000010486 polyoxyethylene sorbitan monolaurate Nutrition 0.000 description 1
- 229920001282 polysaccharide Polymers 0.000 description 1
- 239000005017 polysaccharide Substances 0.000 description 1
- 150000004804 polysaccharides Chemical class 0.000 description 1
- 229920002223 polystyrene Polymers 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- 239000011775 sodium fluoride Substances 0.000 description 1
- 235000013024 sodium fluoride Nutrition 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
Landscapes
- Immobilizing And Processing Of Enzymes And Microorganisms (AREA)
Abstract
Description
【発明の詳細な説明】
［産業上の利用分野］
本発明は、抗原−抗体反応を行なうプレートの表面に微
生物を固相化する方法に間する。DETAILED DESCRIPTION OF THE INVENTION [Industrial Field of Application] The present invention relates to a method of immobilizing microorganisms on the surface of a plate for performing an antigen-antibody reaction.
［従来の技術］
マイクロプレート、プラスチックビーズ、プラスチック
試験管等は、酵素免疫測定法において、抗原な固相化し
、検体中の抗体を測定するための菌として有用である。[Prior Art] Microplates, plastic beads, plastic test tubes, and the like are useful as bacteria for immobilizing antigens and measuring antibodies in specimens in enzyme immunoassay methods.
［発明が解決しようとする課題］
然しなから、抗原が細菌、真菌、原虫等の微生物である
時、微生物は上記器具に固相化されにくく、ブランクが
高い、感度が悪いという問題点かある。[Problems to be solved by the invention] However, when the antigen is a microorganism such as bacteria, fungi, or protozoa, the microorganism is difficult to immobilize on the above-mentioned instrument, resulting in problems such as high blanking and poor sensitivity. .
そこで従来、検体中の微生物に対する抗体価を測定する
場合には、酵素免疫測定法によることなく、微生物懸濁
液を用いた凝集反応法が採用されている。凝集反応法は
、酵素免疫測定法に比較し、感度的に劣る、擬反応が起
こり易い、光学的測定か困難等の欠点を有している。Therefore, conventionally, when measuring the antibody titer against microorganisms in a specimen, an agglutination reaction method using a microorganism suspension has been adopted, instead of an enzyme immunoassay method. The agglutination reaction method has disadvantages compared to the enzyme immunoassay method, such as being inferior in sensitivity, prone to false reactions, and difficult to perform optical measurements.
従って、前記器具に微生物を効率的に固相化てきるので
あれば、検体中の微生物に対する抗体価の測定を、凝集
反応法よりも優れた酵素免疫測定法を利用して行なうこ
とができる。Therefore, if the microorganisms can be efficiently immobilized on the device, the antibody titer against the microorganisms in the specimen can be measured using the enzyme immunoassay method, which is superior to the agglutination reaction method.
本発明は、抗原−抗体反応を行なう器具の表面に微生物
を効率的に固相化することを目的とする。An object of the present invention is to efficiently immobilize microorganisms on the surface of a device that performs an antigen-antibody reaction.
［課題を解決するための手段］
請求項１に記載の本発明は、抗原−抗体反応を行なう器
具の表面に微生物を固相化する方法において、該器具の
表面に遊離アミノ基を有する高分子溶液を接触、吸着せ
しめ、該高分子溶液を除去、洗浄後、遊離アミノ基と化
学的に結合する官能基な分子内に２つ以上もつ架橋剤溶
液を接触、反応させ、該架橋剤溶液を除去、洗浄後、微
生物溶液を接触、反応させ、該微生物溶液を除去、洗浄
するようにしたものである。[Means for Solving the Problems] The present invention as set forth in claim 1 provides a method for immobilizing microorganisms on the surface of a device for performing an antigen-antibody reaction. After contacting and adsorbing the solution, removing and washing the polymer solution, contacting and reacting with a crosslinking agent solution having two or more functional groups in the molecule that chemically binds to free amino groups, and causing the crosslinking agent solution to react. After removal and washing, the microorganism solution is contacted and reacted, and the microorganism solution is removed and washed.
請求項２に記載の本発明は、前記高分子がＰｏ１ｙ−Ｌ
−Ｌｙｓｉｎｅであるようにしたものである。In the present invention according to claim 2, the polymer is Poly-L.
-Lysine.
［作用］
請求項１に記載の本発明によれば、下記■の作用がある
。[Action] According to the present invention as set forth in claim 1, there is the following effect (2).
■酵素免疫測定法において、通常、抗原を固相化し、反
応を行なうためのマイクロプレート、ビーズ、試験管等
の器具は、プラスチック、主としてポリスチレンを成形
して作られる。そして、これらの器具はその表面に蛋白
質を吸着する性質を有する。他方、細菌、真菌、原虫等
の微生物はその外側を多糖からなる細胞壁でおおわれて
おり、これらの器具表面に吸着しない、或いはしにく
し＼ 。■In the enzyme immunoassay method, the instruments such as microplates, beads, test tubes, etc. for immobilizing antigens and carrying out reactions are usually made of molded plastic, mainly polystyrene. These devices have the property of adsorbing proteins on their surfaces. On the other hand, microorganisms such as bacteria, fungi, and protozoa are covered with a cell wall made of polysaccharide, so they do not adsorb to the surface of these devices or are difficult to adhere to.
death\ .
そこで、本発明にあっては、先ず器具表面に吸着する性
質を有している高分子（これは架橋剤との化学的結合性
を有する遊離アミノ基を備えている）を該器具表面に吸
着させた０次に、上記高分子と、固相化しようとする微
生物とを架橋するための架橋剤を反応させた。この時、
架橋剤の一方の反応基は高分子の遊離アミノ基と結合し
、他方の反応基は官能基として残る。従って、微生物を
接触せしめれば、上記架橋剤の残存している官能基か微
生物のアミノ基と反応する結果、器具表面に微生物を効
率良く固相化てきる。Therefore, in the present invention, first, a polymer having the property of adsorbing to the surface of the device (which has free amino groups that have chemical bonding properties with a crosslinking agent) is adsorbed onto the surface of the device. Next, a crosslinking agent for crosslinking the polymer and the microorganism to be solidified was reacted. At this time,
One reactive group of the crosslinking agent binds to the free amino group of the polymer, while the other reactive group remains as a functional group. Therefore, when microorganisms are brought into contact, the remaining functional groups of the crosslinking agent react with the amino groups of the microorganisms, and as a result, the microorganisms are efficiently immobilized on the surface of the instrument.
請求項２に記載の本発明によれば、下記■の作用かある
。According to the present invention as set forth in claim 2, there is the following effect (2).
■前記高分子としてＰｏ１ｙ−Ｌ−Ｌｙｓｉｎｅを用い
たから、器具表面に対する吸着か確実である。(2) Since Poly-L-Lysine was used as the polymer, it is certain that it is adsorbed onto the surface of the instrument.
尚、本発明の実施において、前記高分子としては、Ｐｏ
１ｙ−Ｌ−Ｌｙｓｉｎｅの他、血清アルブミン、アルギ
ン酸を用いることができる。In the practice of the present invention, as the polymer, Po
In addition to 1y-L-Lysine, serum albumin and alginic acid can be used.
又、本発明の実施において、前記架橋剤としては、グル
タルアルデヒドやビスイミデート系、ジイソシアネート
系の架橋剤を用いることができる。Further, in the practice of the present invention, as the crosslinking agent, a glutaraldehyde, bisimidate, or diisocyanate type crosslinking agent can be used.
［実施例コ
（実施例１）
本発明を用いて、Ａ群連釦球菌に対するウサギ抗血清に
抗体価を測定した。[Example 1 (Example 1) Using the present invention, the antibody titer was measured for rabbit antiserum against group A Streptococcus.
（Ａ） Ａ群連鎖球菌固相化マイクロプレートの作製
■９６穴マイクロプレートの各ウェルに、１０μｇ／ａ
ｎのＰｏ１ｙ−Ｌ−Ｌｙｓｉｎｅ炭ＷＩＩＩＩ衝液（Ｐ
Ｈ９，ｌ１ｔ）を１００μβずつ分注し、遊離アミノ基
を有するＰｏ１ｙ−Ｌ−Ｌｙｓｉｎｅを接触、吸着せし
める。室温にて３０分間靜１した後、各ウェル内液を除
去、０．１５ＭＮａＣｌ２を含むリン酸緩衝液（ＰＨ７
，４） （以下ＰＢＳと略記する）で洗浄する。(A) Preparation of group A streptococcus-immobilized microplate ■ 10 μg/a in each well of a 96-well microplate
Poly-L-Lysine charcoal WIII buffer solution (P
H9, l1t) is dispensed in 100 μβ portions and brought into contact with and adsorbed with Poly-L-Lysine having a free amino group. After incubating at room temperature for 30 minutes, the solution in each well was removed and diluted with phosphate buffer (PH7) containing 0.15M NaCl2.
, 4) (hereinafter abbreviated as PBS).
■次に、上述の各ウェルに、架橋剤としての０．２５％
グルタルアルデヒド−ＰＢＳ溶液を １００μβ／ウエ
ルずつ分注し、接触、反応せしめる。室温にて３０分間
靜１した後、各ウェル内液を除去、ＰＢＳで洗浄する。■Next, add 0.25% as a crosslinking agent to each well mentioned above.
A glutaraldehyde-PBS solution is dispensed at 100 μβ/well, and brought into contact and reacted. After incubating at room temperature for 30 minutes, the solution in each well was removed and washed with PBS.
０次に、Ａ群連錆球菌をＰＢＳに浮遊させた懸濁液（約
１０？個７１０を１００μβずつ各ウェルに分注し、接
触、反応せしめる。室温にて３０分間放置した後、各ウ
ェル内懸濁液を除去、ＰＢＳで洗浄する。0 Next, a suspension of group A streptococci (approximately 10 710 particles of 100μβ) is dispensed into each well and brought into contact and reacted. After standing at room temperature for 30 minutes, each well is Remove the internal suspension and wash with PBS.
■クリシンーＢＳＡ （ウシ血清アルブミン）溶液を
各ウェルに １００μβずつ分注し、室温にて３０分間
靜１し、遊離のアルデヒド基をマスクする。各ウェルを
ＰＢＳにて洗浄する。■ Dispense 100 μβ of chrysin-BSA (bovine serum albumin) solution to each well and incubate at room temperature for 30 minutes to mask free aldehyde groups. Wash each well with PBS.
ＣＢ）ウサギ抗血清の抗体価測定
■Ａ群連鎖球菌で免疫したウサギの抗血清を、０．５Ｍ
塩化ナトリウム、 ０．１％ＢＳＡ、 ０．５％Ｔｗ
ｅｅｎ２０を含む１０ｍＭリン酸緩衝液て２倍ずつ段階
希釈する。CB) Antibody titer measurement of rabbit antiserum ■ Antiserum from a rabbit immunized with group A streptococcus was added to 0.5M
Sodium chloride, 0.1%BSA, 0.5%Tw
Serially dilute 2-fold in 10 mM phosphate buffer containing een20.
■上記（Ａ）の本発明方法にて作製した９６穴マイクロ
プレートの各ウェルに、各希釈抗血清を１００μβずつ
分取する。室温にて１時間静置の後、各ウェル内液を除
去、０．０５％Ｔｖｅｅｎ２０を含むＰＢＳにて洗浄す
る。(2) Aliquot 100 μβ of each diluted antiserum into each well of the 96-well microplate prepared by the method of the present invention described in (A) above. After standing at room temperature for 1 hour, the solution in each well was removed and washed with PBS containing 0.05% Tveen20.
■パーオキシダーゼ標識抗つサギーＩｇＧ抗体（ヒツジ
由来）溶液を、各ウェルに１００μρずつ分注し、室温
にて１時間静置する。(2) Dispense 100 μρ of peroxidase-labeled anti-Saggy IgG antibody (derived from sheep) solution to each well and leave it at room temperature for 1 hour.
■各つェルを０．０５％Ｔｗｅｅｎ２０を含むＰＢＳに
て洗浄し、各ウェルに２．２−ａｚｉｎｏ−ｄｉ−［３
−ｅｔｈｙｌ−ｂｅｎｚｔｈｉａｚｏｌｉｎ ５ｕｌｐ
ｈｏｎｉｃ ａｃｉｄｌ ０．１５ｍｇ／ｍＡ、３０％
ＨＪｔ ５ｕ Ａ ／１０ｍＡを含ムｏ、ＩＭクエン
酸／ Ｎａ０Ｈ（ＰＨ４，０）溶液を２００μρずつ分
注する。室温にて３０分間靜装の後、 １Ｍフッ化ナト
リウム液１００μβを加え、反応を停止させ、４１５ｒ
＋＋ｗの吸光度を測定する（第１図参照）。■ Wash each well with PBS containing 0.05% Tween20, and add 2.2-azino-di-[3
-ethyl-benzthiazolin 5ulp
honic acidl 0.15mg/mA, 30%
Dispense 200 μρ of IM citric acid/NaOH (PH 4,0) solution containing HJt 5u A /10 mA. After incubating at room temperature for 30 minutes, 100μβ of 1M sodium fluoride solution was added to stop the reaction, and 415r
Measure the absorbance of ++w (see Figure 1).
尚、上記（Ｂ）■で用いるマイクロプレートとして、Ｐ
ｏ１ｙ−Ｌ−Ｌｙｓｉｎｅのみを処理したプレートに菌
体を固相化したもの、及びｐｏｌｙ−１−Ｌｙｓｉｎｅ
とグルタルアルデヒドの両処理をしないプレートに菌体
を固相化したものを用いた場合の測定結果を第１図に併
せ示した。In addition, as the microplate used in (B)■ above, P
A plate in which bacterial cells were immobilized on a plate treated with only o1y-L-Lysine, and a plate treated with poly-1-Lysine.
Figure 1 also shows the measurement results when using a plate with immobilized bacterial cells that was not treated with either glutaraldehyde or glutaraldehyde.
第１図によれば、本発明の実施により、マイクロプレー
ト表面に菌体な効率良く固相化できたことが認められる
。According to FIG. 1, it is recognized that by implementing the present invention, bacterial cells could be efficiently immobilized on the surface of the microplate.
（Ｃ） ＩＩ集反応によるウサギ抗血清の抗体価測定Ａ
群連鎖球菌浮遊液（約１０８個／ｌΩ）とウサギ抗血清
希釈液をスライドガラス上て１滴ずつ適下、混合し、凝
集の有無を調べた。(C) Antibody titer measurement A of rabbit antiserum by II collection reaction
A suspension of group streptococci (approximately 108 cells/lΩ) and a diluted rabbit antiserum solution were dropped drop by drop onto a slide glass and mixed, and the presence or absence of agglutination was examined.
Ａ群連鎖球菌抗血清の抗体価測定において、前記（Ｂ）
の本発明を用いた酵素免疫測定法による結果と、上記（
Ｃ）の凝集反応法による結果とを表１に比較して示した
。In measuring the antibody titer of group A streptococcal antiserum, the above (B)
The results of enzyme immunoassay using the present invention and the above (
Table 1 shows a comparison with the results obtained by the agglutination reaction method in C).
表１によれば、本発明を用いた酵素免疫測定法の実施に
より、抗体価を極めて高感度で測定できることが認めら
れる。According to Table 1, it is recognized that antibody titers can be measured with extremely high sensitivity by implementing the enzyme immunoassay method using the present invention.
（実施例２）
本発明を用いて、Ｃａｎｄｉｄａ ａｌｂｉｃａｎｓに
対するウサギ抗血清の抗体価を測定した。(Example 2) Using the present invention, the antibody titer of rabbit antiserum against Candida albicans was measured.
（Ａ） Ｃａｎｄｉｄａ ａｌｂｉｃａｎｓ 固相化
マイクロプレートの作製
前記実施例１の（Ａ）におけると同様にして、Ｃａｎｄ
ｉｄａ ａｌｂｉｃａｎｓ 固相化マイクロプレート
を作製した。(A) Preparation of Candida albicans solid-phase microplate In the same manner as in (A) of Example 1, Candida albicans
ida albicans solid-phase microplate was prepared.
（Ｂ）ウサギ抗血清の抗体価測定
前記実施例１の（Ｂ）におけると同様にして、ウサギ抗
血清の抗体価を測定し、第２図を得た。(B) Measurement of antibody titer of rabbit antiserum The antibody titer of rabbit antiserum was measured in the same manner as in (B) of Example 1 above, and FIG. 2 was obtained.
尚、ｐｏｌｙ−Ｌ−Ｌｙｓｉｎｅとグルタルアルデヒド
の両処理をしないプレートに菌体を固相化したものを用
いた場合の測定結果を、第２図に併せ示した。Incidentally, the measurement results when using a plate in which bacterial cells were immobilized on a plate that was not treated with both poly-L-Lysine and glutaraldehyde are also shown in FIG.
第２図によれば、本発明の実施により、マイクロプレー
ト表面に菌体を効率良く固相化できたことが認められる
。According to FIG. 2, it is recognized that by implementing the present invention, bacterial cells could be efficiently immobilized on the surface of the microplate.
（Ｃ）凝集反応によるウサギ抗血清の抗体価測定前記実
施例１の（Ｃ）におけると同様にして、凝集反応による
ウサギ抗血清の抗体価を測定した。(C) Measurement of antibody titer of rabbit antiserum by agglutination reaction The antibody titer of rabbit antiserum was measured by agglutination reaction in the same manner as in (C) of Example 1 above.
Ｃａｎｄｉｄａ ａｌｂｉｃａｎｓ抗血清の抗体価測定
におし＼て、前記ＣＢ）の本発明を用いた酵素免疫測定
法による結果と、上記（Ｃ）の凝集反応法による結果と
を表２に比較して示した。In measuring the antibody titer of Candida albicans antiserum, Table 2 shows a comparison between the results of the enzyme immunoassay method using the present invention in CB) and the agglutination reaction method in (C) above. Ta.
表２によれば、本発明を用いた酵素免疫測定法の実施に
より、抗体価を極めて高感度で測定てきることが認めら
れる。According to Table 2, it is recognized that antibody titers can be measured with extremely high sensitivity by implementing the enzyme immunoassay method using the present invention.
［発明の効果］
以上のように本発明によれば、抗原−抗体反応を行なう
器具の表面に微生物を効率的に固相化することがてきる
。[Effects of the Invention] As described above, according to the present invention, microorganisms can be efficiently immobilized on the surface of a device that performs an antigen-antibody reaction.
第１図はＡ群連鎖球菌抗血清の抗体価測定結果を示す線
図、第２図はＣａｎｄｉｄａ ａｌｂｉｃａｎｓ抗血清
の抗体価測定結果を示す線図である。FIG. 1 is a diagram showing the results of measuring the antibody titer of group A streptococcus antiserum, and FIG. 2 is a diagram showing the results of measuring the antibody titer of Candida albicans antiserum.
Claims (2)
相化する方法において、該器具の表面に遊離アミノ基を
有する高分子溶液を接触、吸着せしめ、該高分子溶液を
除去、洗浄後、遊離アミノ基と化学的に結合する官能基
を分子内に２つ以上もつ架橋剤溶液を接触、反応させ、
該架橋剤溶液を除去、洗浄後、微生物溶液を接触、反応
させ、該微生物溶液を除去、洗浄することを特徴とする
微生物の固相化方法。(1) In a method of immobilizing microorganisms on the surface of a device for performing an antigen-antibody reaction, the surface of the device is brought into contact with and adsorbed with a polymer solution having free amino groups, and the polymer solution is removed and washed. , contacting and reacting with a crosslinking agent solution having two or more functional groups in the molecule that chemically bond with free amino groups,
A method for immobilizing microorganisms, which comprises removing and washing the crosslinking agent solution, contacting and reacting with a microorganism solution, and removing and washing the microorganism solution.
請求項１記載の微生物の固相化方法。(2) The method for immobilizing microorganisms according to claim 1, wherein the polymer is Poly-L-Lysine.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP8109490A JPH03280881A (en) | 1990-03-30 | 1990-03-30 | Method for immobilizing microorganism |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP8109490A JPH03280881A (en) | 1990-03-30 | 1990-03-30 | Method for immobilizing microorganism |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| JPH03280881A true JPH03280881A (en) | 1991-12-11 |
Family
ID=13736803
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP8109490A Pending JPH03280881A (en) | 1990-03-30 | 1990-03-30 | Method for immobilizing microorganism |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPH03280881A (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH07318561A (en) * | 1994-05-24 | 1995-12-08 | Daiichi Rajio Isotope Kenkyusho:Kk | Biochemical fine particle, production thereof and immunoassay |
| WO2004108918A1 (en) * | 2003-06-05 | 2004-12-16 | Sony Corporation | Immobilization support, process for producing the same, electrode, process for producing the same, electrode reaction utilizing apparatus and process for producing the same |
| JP2008545470A (en) * | 2005-05-24 | 2008-12-18 | ステリス インコーポレイテッド | Biological indicator |
-
1990
- 1990-03-30 JP JP8109490A patent/JPH03280881A/en active Pending
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH07318561A (en) * | 1994-05-24 | 1995-12-08 | Daiichi Rajio Isotope Kenkyusho:Kk | Biochemical fine particle, production thereof and immunoassay |
| WO2004108918A1 (en) * | 2003-06-05 | 2004-12-16 | Sony Corporation | Immobilization support, process for producing the same, electrode, process for producing the same, electrode reaction utilizing apparatus and process for producing the same |
| JP2008545470A (en) * | 2005-05-24 | 2008-12-18 | ステリス インコーポレイテッド | Biological indicator |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| US4184849A (en) | Mixed agglutination | |
| US4495296A (en) | Labeled anti-hapten antibodies and their use as a universal reagent for solid phase radio- and/or enzyme-immunoassays | |
| US4670383A (en) | Immune-chemical measurement process for haptens and proteins | |
| US5017474A (en) | Wash solution, test kit and method for the determination of an immunological ligand | |
| JPH0731197B2 (en) | Lower alcohol sulphate washing solution, test kit and method for measuring immunoligand | |
| US20070184504A1 (en) | Assay for anti-INGAP antibodies | |
| JPS6217188B2 (en) | ||
| JP3186831B2 (en) | Aggregation complex as blood group reagent | |
| JPH03280881A (en) | Method for immobilizing microorganism | |
| JP3282129B2 (en) | Solid phase non-separable enzyme analysis | |
| JP2736058B2 (en) | Manufacturing method of immunoassay device | |
| JPH04236353A (en) | Measuring method for antibody | |
| JPH03170058A (en) | Reagent complex for immunoassay | |
| EP0291479A1 (en) | Immunoassay method for the diagnosis of chlamydia infection | |
| WO1995002184A1 (en) | Water soluble polymers for use in immunoassays and dna hybridization assays | |
| WO1987007956A1 (en) | Preparation for immunoassay and method for producing the preparation | |
| US5185128A (en) | Test kit containing labeled receptor and wash solution for determination of an immunological ligand | |
| CN117949668A (en) | Detection reagent and kit for alpha tumor necrosis factor | |
| JPH0389166A (en) | Method for measuring antibody activity | |
| JPH02259569A (en) | Method for measuring rheumatism factor | |
| JP3936678B2 (en) | Anti-syphilis treponema antibody measurement reagent and anti-syphilis treponema antibody measurement method | |
| CN114924065A (en) | Detection kit for pepsinogen I as well as preparation method and application thereof | |
| JPS63151856A (en) | Immunological measurement reagent using monoclonal antibody for human protein s | |
| JPS63235868A (en) | Determination of rheumatism factor | |
| EP0133553A2 (en) | Reagent and process for the quantitative determination of antibodies |