JPH055738A - Determining method of free haptoglobin - Google Patents
Determining method of free haptoglobinInfo
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- JPH055738A JPH055738A JP18207691A JP18207691A JPH055738A JP H055738 A JPH055738 A JP H055738A JP 18207691 A JP18207691 A JP 18207691A JP 18207691 A JP18207691 A JP 18207691A JP H055738 A JPH055738 A JP H055738A
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- Prior art keywords
- haptoglobin
- type
- antibody
- free
- monoclonal
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Abstract
Description
【0001】[0001]
【産業上の利用分野】本発明は遊離ハプトグロビンの定
量方法に関する。FIELD OF THE INVENTION The present invention relates to a method for quantifying free haptoglobin.
【0002】[0002]
【従来技術】ハプトグロビン(Hp)は、血清α2−グ
ロブリンに属する糖タンパクで、溶血によって生ずるヘ
モグロビン(Hb)と強く結合して、安定な複合体を形
成する。Hp−Hb複合体形成によるマイクロ分子化
は、Hbの腎臓からの流失を防ぐだけではなく、Hbに
よる腎細尿管障害を防止する効果もある。Hp−Hb複
合体は、網内系細胞により、摂取、処理される。2. Description of the Related Art Haptoglobin (Hp) is a glycoprotein belonging to serum α 2 -globulin and strongly binds to hemoglobin (Hb) produced by hemolysis to form a stable complex. Micromolecularization by formation of Hp-Hb complex not only prevents Hb from being washed away from the kidney, but also has an effect of preventing renal tubular tract injury caused by Hb. The Hp-Hb complex is taken up and processed by reticuloendothelial cells.
【0003】ハプトグロビンには、Hp1−1型(分子
量約 10万)、Hp2−1型(分子量約40万)、H
p2−1型(分子量約22万)の3種の血清型が存在
し、更に、6種の亜型が存在する。従来、Mancin
i法により総ハプトグロビンを求める際には、3種の血
清型により、ハプトグロビンの拡散速度が異なる為、実
測値にファクターを掛けて値を算出する必要があり、直
接、総ハプトグロビンを定量することは困難だった。ま
た、従来は、遊離ハプトグロビンを直接定量する方法は
存在しなかった。Haptoglobin includes Hp1-1 type (molecular weight of about 100,000), Hp2-1 type (molecular weight of about 400,000), and Hp2-1 type.
There are 3 serotypes of p2-1 type (molecular weight about 220,000), and 6 subtypes. Traditionally, Mancin
When the total haptoglobin is determined by the i method, the diffusion rate of haptoglobin differs depending on the three serotypes, so it is necessary to multiply the measured value by a factor to calculate the value. It is not possible to directly quantify total haptoglobin. It was difficult. Further, conventionally, there has been no method for directly quantifying free haptoglobin.
【0004】本発明者は、担体に固定化した遊離ヘモグ
ロビンに遊離ハプトグロビンを結合させ、Hp−Hb複
合体を形成させた後、更に、酵素標識ハプトグロビン抗
体を結合させ、発色する方法が、遊離ハプトグロビンの
定量に最適であることを見出し、本発明を完成した。The present inventor has proposed a method of binding free haptoglobin to free hemoglobin immobilized on a carrier to form an Hp-Hb complex, and then further binding an enzyme-labeled haptoglobin antibody to develop color. The present invention has been completed by finding that it is optimum for the quantification of
【0005】[0005]
【実施例】以下に実施例を挙げて、本発明を更に詳細に
説明するが、本発明は以下の実施例には何ら限定される
ものではない。The present invention will be described in more detail with reference to the following examples, but the present invention is not limited to the following examples.
【0006】実施例1 A.遊離ハプトグロビン定量キットの構成 (1)遊離ヘモグロビンを固定化したアガロースを封入
したカートリッジ (2)標準遊離ハプトグロビン (3)酵素標識遊離ハプトグロビン抗体(モノクローナ
ル1−1 型抗体) 酵素標識遊離ハプトグロビン抗体(モノクローナル2−
1 型抗体) 酵素標識遊離ハプトグロビン抗体(モノクローナル2−
2 型抗体) (4)洗浄液 (5)酵素基質 (6)緩衝液 (7)反応停止液 B.測定操作 (1)小試験管に、検体1ml(1検体につき3本)と
遊離ヘモグロビンを固定化したアガロースを封入レたカ
ートリッジ各1個を分取、混和し、室温で30分間イン
キュベートした。 (2)各試験管の内容液を捨て、試験管内の遊離ヘモダ
ロビンを固定化したアガロースを封入したカートリッジ
を洗浄液2mlで2回洗浄、水切りをした。 (3)(2)の操作をした各試験管(1検体につき3
本)に、3種類の酵素標識遊離ハプトグロビン抗体(モ
ノクローナル1−1型抗体または、モノクローナル2−
1型抗体、または、モノクローナル2−2型抗体)2m
lを別々に添加、混和し、37℃で30分間インキュベ
ートした。 (4)(3)の操作をした各試験管の内容液を捨て、試
験管内の遊離ヘモグロビンを固定化したアガロースを封
入したカートリッジを洗浄液2mlで3回洗浄、水切り
をした。 (5)(4)の操作をした各試験管に酵素基質液500
ulを添加、混和し、37℃で30分間インキュベート
した。 (6)(5)の操作をした各試験管に反応停止液2ml
を添加、混和した。 (7)ブランクを対照にして、492nmの吸光度を測
定した。 (8)標準遊離ハプトグロビンを用いて同一操作をして
描いた3種類(1−1型、2−1型、2−2型)の検量
線から検体の遊離ハプトグロビン量を求めた。Example 1 A. Composition of Free Haptoglobin Quantification Kit (1) Cartridge encapsulating agarose with immobilized free hemoglobin (2) Standard free haptoglobin (3) Enzyme-labeled free haptoglobin antibody (monoclonal type 1-1 antibody) Enzyme-labeled free haptoglobin antibody (monoclonal 2) −
Type 1 antibody) Enzyme-labeled free haptoglobin antibody (monoclonal 2-
Type 2 antibody) (4) Wash solution (5) Enzyme substrate (6) Buffer solution (7) Reaction stop solution B. Measurement operation (1) 1 ml of each sample (3 per 1 sample) and 1 cartridge each containing agarose on which free hemoglobin was immobilized were placed in a small test tube, mixed, and incubated at room temperature for 30 minutes. (2) The content liquid in each test tube was discarded, and the cartridge containing agarose on which free hemodalobin was immobilized was washed twice with 2 ml of the washing solution and drained. (3) Each test tube operated in (2) (3 per sample)
3) enzyme-labeled free haptoglobin antibody (monoclonal type 1-1 antibody or monoclonal 2-
Type 1 antibody or monoclonal type 2-2 antibody) 2m
1 was added separately, mixed, and incubated at 37 ° C. for 30 minutes. (4) The content liquid of each test tube subjected to the operation of (3) was discarded, and the cartridge containing agarose in which free hemoglobin was immobilized was washed with 2 ml of the washing solution three times and drained. (5) The enzyme substrate solution 500 is placed in each of the test tubes subjected to the operations of (4).
ul was added, mixed, and incubated at 37 ° C. for 30 minutes. (6) 2 ml of reaction stop solution in each test tube operated in (5)
Was added and mixed. (7) Using the blank as a control, the absorbance at 492 nm was measured. (8) The amount of free haptoglobin of the sample was determined from the calibration curves of three types (1-1 type, 2-1 type, 2-2 type) drawn by performing the same operation using standard free haptoglobin.
【0007】実施例2. A.遊離ハプトグロビン定量キットの構成 (1)反応性官能基を固相化し、更に遊離ヘモグロビン
を共有結合させたビーズ (2)標準遊離ハプトグロビン (3)酵素標識遊離ハプトグロビン抗体(モノクローナ
ル1−1 型抗体) 酵素標識遊離ハプトグロビン抗体(モノクローナル2−
1 型抗体) 酵素標識遊離ハプトグロビン抗体(モノクローナル2−
2 型抗体) (4)洗浄液 (5)酵素基質 (6)緩衝液 (7)反応停止液 B.測定操作 (1)小試験管に、検体1ml(1検体につき 3本)
と遊離ヘモグロビンを共有結合させたビーズ各1個を分
取、混和し、室温で30分間インキュベートした。 (2)各試験管の内容液を捨て、試験管内の遊離ヘモグ
ロビンを共有結合させたビーズを洗浄液2mlで2回洗
浄した。 (3)(2)の操作をした各試験管(1検体につき3
本)に、3種類の酵素標識遊離ハプトグロビン抗体(モ
ノクローナル1−1 型抗体または、モノクローナル2
−1型抗体、または、モノクローナル 2−2 型抗
体)2mlを別々に添加、混和し、37℃で30分間イ
ンキュベートした。 (4)(3)の操作をした各試験管の内容液を捨て、試
験管内の遊離ヘモグロビンを共有結合させたビーズを洗
浄液2mlで3回洗浄した。 (5)新たに準備した各試験管に酵素基質液500ul
を秤量し、(4)の操作をしたビーズを移し、混和後、
37℃で30分間インキュベートした。 (6)(5)の操作をした各試験管に反応停止液2ml
を添加、混和した。 (7)ブランクを対照にして、492nmの吸光度を測
定した。 (8)標準遊離ハプトグロビンを用いて同一操作をして
描いた3種類(1−1型、2−1型、2−2型)の検量
線から検体の遊離ハプトグロビン量を求めた。Example 2. A. Composition of Free Haptoglobin Quantification Kit (1) Beads on which reactive functional groups are immobilized and further covalently bound to free hemoglobin (2) Standard free haptoglobin (3) Enzyme-labeled free haptoglobin antibody (monoclonal 1-1 antibody) enzyme Labeled free haptoglobin antibody (monoclonal 2-
Type 1 antibody) Enzyme-labeled free haptoglobin antibody (monoclonal 2-
Type 2 antibody) (4) Wash solution (5) Enzyme substrate (6) Buffer solution (7) Reaction stop solution B. Measurement operation (1) 1 ml sample (3 per sample) in a small test tube
And 1 bead to which free hemoglobin was covalently bound were collected, mixed, and incubated at room temperature for 30 minutes. (2) The content liquid of each test tube was discarded, and the beads in the test tube to which the free hemoglobin was covalently bound were washed twice with 2 ml of the washing solution. (3) Each test tube operated in (2) (3 per sample)
3) enzyme-labeled free haptoglobin antibody (monoclonal type 1-1 antibody or monoclonal 2
-1 type antibody or 2 ml of monoclonal 2-2 type antibody) was added separately, mixed, and incubated at 37 ° C for 30 minutes. (4) The content liquid of each test tube subjected to the operation of (3) was discarded, and the beads in the test tube to which the free hemoglobin was covalently bound were washed 3 times with 2 ml of the washing solution. (5) Add 500ul of enzyme substrate solution to each newly prepared test tube.
Is weighed, and the beads obtained in step (4) are transferred and mixed,
Incubated at 37 ° C for 30 minutes. (6) 2 ml of reaction stop solution in each test tube operated in (5)
Was added and mixed. (7) Using the blank as a control, the absorbance at 492 nm was measured. (8) The amount of free haptoglobin of the sample was determined from the calibration curves of three types (1-1 type, 2-1 type, 2-2 type) drawn by performing the same operation using standard free haptoglobin.
【0008】実施例3. A.遊離ハプトグロビン定量キットの構成 (1)遊離ヘモグロビンを共有結合させたマイクロタイ
タープレート (2)標準遊離ヘモグロビン (3)酵素標識遊離ハプトグロビン抗体(モノクローナ
ル1−1 型抗体) 酵素標識遊離ハプトグロビン抗体(モノクローナル2−
1 型抗体) 酵素標識遊離ハプトグロビン抗体(モノクローナル2−
2 型抗体) (4)洗浄液 (5)酵素基質 (6)緩衝液 (7)反応停止液 B.測定操作 (1)遊離ヘモグロビンを共有結合させたマイクロタイ
タープレートに、検体200ulを秤量、混和し、室温
で30分間インキュベートした。 (2)各マイクロタイタープレートの内容液を捨て、各
マイクロタイタープレートを洗浄液300ulで3回洗
浄した。 (3)(2)の操作をしたマイクロタイタープレートの
各穴(1検体につき3穴に、3種類の酵素標識遊離ハプ
トグロビン抗体(モノクローナル1−1型抗体または、
モノクローナル2−1型抗体、または、モノクローナル
2−2型抗体)200ulを別々に添加、混和し、37
℃で30分間インキュベートした。 (4)(3)の操作をした各マイクロタイタープレート
の内容液を捨て、各試験管を洗浄液300ulで3回洗
浄した。 (5)(4)の操作をした各マイクロタイタープレート
に酵素基質液200ulを添加、混和し、37℃で30
分間インキュベートした。 (6)(5)の操作をした各試験管に反応停止液100
ulを添加、混和した。 (7)ブランクを対照にして、492nmの吸光度を測
定した。 (8)標準遊離ハプトグロビンを用いて同一操作をして
描いた3種類(1−1型、2−1型、2−2型)の検量
線から検体の遊離ハプトグロビン量を求めた。Embodiment 3. A. Structure of Free Haptoglobin Assay Kit (1) Microtiter plate to which free hemoglobin was covalently bound (2) Standard free hemoglobin (3) Enzyme-labeled free haptoglobin antibody (monoclonal type 1-1 antibody) Enzyme-labeled free haptoglobin antibody (monoclonal 2-
Type 1 antibody) Enzyme-labeled free haptoglobin antibody (monoclonal 2-
Type 2 antibody) (4) Wash solution (5) Enzyme substrate (6) Buffer solution (7) Reaction stop solution B. Measurement operation (1) 200 μl of the sample was weighed and mixed in a microtiter plate to which free hemoglobin was covalently bonded, and the mixture was incubated at room temperature for 30 minutes. (2) The content liquid of each microtiter plate was discarded, and each microtiter plate was washed 3 times with 300 ul of the washing liquid. (3) Each hole of the microtiter plate subjected to the operation of (2) (three kinds of one sample, three kinds of enzyme-labeled free haptoglobin antibody (monoclonal type 1-1 antibody, or
Monoclonal 2-1 type antibody or monoclonal 2-2 type antibody) 200 ul are separately added and mixed, and 37
Incubated at 0 ° C for 30 minutes. (4) The content liquid of each microtiter plate subjected to the operation of (3) was discarded, and each test tube was washed 3 times with 300 ul of the washing liquid. (5) Add 200 ul of enzyme substrate solution to each microtiter plate obtained in the procedure of (4), mix, and mix at 37 ° C. for 30 minutes.
Incubated for minutes. (6) The reaction stop solution 100 is added to each of the test tubes operated in (5).
ul was added and mixed. (7) Using the blank as a control, the absorbance at 492 nm was measured. (8) The amount of free haptoglobin of the sample was determined from the calibration curves of three types (1-1 type, 2-1 type, 2-2 type) drawn by performing the same operation using standard free haptoglobin.
【0009】実施例4. A.遊離ハプトグロビン定量キットの構成 (1)反応性官能基に遊離ヘモグロビンを共有結合させ
た試験管 (2)標準遊離ハプトグロビン (3)酵素標識遊離ハプトグロビン抗体(モノクローナ
ル1−1 型抗体) 酵素標識遊離ハプトグロビン抗体(モノクローナル2−
1 型抗体) 酵素標識遊離ハプトグロビン抗体(モノクローナル2−
2 型抗体) (4)洗浄液 (5)酵素基質 (6)緩衝液 (7)反応停止液 B.測定操作 (1)反応性官能基を固体相化し、更に遊離ヘモグロビ
ンを共有結合させた試験管に、検体1mlを秤量、混和
し、室温で30分間インキュベートした。 (2)各試験管の内容液を捨て、各試験管を洗浄液3m
lで3回洗浄した。 (3)(2)の操作をした各試験管(1検体につき3
本)に、3種類の酵素標識遊離ハプトグロビン抗体(モ
ノクローナル1−1型抗体または、モノクローナル2−
1型抗体、または、モノクローナル2−2 型抗体)2
mlを別々に添加、混和し、37℃で30分間インキュ
ベートした。 (4)(3)の操作をした各試験管の内容液を捨て、各
試験管を洗浄液3mlで3回洗浄した。 (5)(4)の操作をした各試験管に酵素基質液1ml
を添加、混和し、37℃で30分間インキュベートし
た。 (6)(5)の操作をした各試験管に反応停止液1ml
を添加、混和した。 (7)ブランクを対照にして、492nmの吸光度を測
定した。 (8)標準遊離ハプトグロビンを用いて同一操作をして
描いた3種類(1−1型、2−1型、2−2型)の検量
線から検体の遊離ハプトグロビン量を求めた。Example 4. A. Composition of Free Haptoglobin Quantitative Kit (1) Test tube with free hemoglobin covalently bound to reactive functional group (2) Standard free haptoglobin (3) Enzyme-labeled free haptoglobin antibody (monoclonal type 1-1 antibody) Enzyme-labeled free haptoglobin antibody (Monoclonal 2-
Type 1 antibody) Enzyme-labeled free haptoglobin antibody (monoclonal 2-
Type 2 antibody) (4) Wash solution (5) Enzyme substrate (6) Buffer solution (7) Reaction stop solution B. Measurement procedure (1) 1 ml of a sample was weighed and mixed in a test tube in which a reactive functional group was solid-phased and free hemoglobin was covalently bonded, and the mixture was incubated at room temperature for 30 minutes. (2) Discard the contents of each test tube and wash each test tube with 3m of washing liquid.
Wash 3 times with 1. (3) Each test tube operated in (2) (3 per sample)
3) enzyme-labeled free haptoglobin antibody (monoclonal type 1-1 antibody or monoclonal 2-
Type 1 antibody or monoclonal 2-2 type antibody) 2
ml was added separately, mixed, and incubated at 37 ° C for 30 minutes. (4) The liquid content of each test tube subjected to the operation of (3) was discarded, and each test tube was washed 3 times with 3 ml of the washing solution. (5) 1 ml of enzyme substrate solution in each test tube that was operated in (4)
Was added, mixed, and incubated at 37 ° C. for 30 minutes. (6) 1 ml of reaction stop solution in each test tube operated in (5)
Was added and mixed. (7) Using the blank as a control, the absorbance at 492 nm was measured. (8) The amount of free haptoglobin of the sample was determined from the calibration curves of three types (1-1 type, 2-1 type, 2-2 type) drawn by performing the same operation using standard free haptoglobin.
【0010】実施例5. A.遊離ハプトグロビン定量キットの構成 (1)遊離ヘモグロビンを固定化したアガロースを封入
したカートリッジ (2)標準遊離ハプトグロビン (3)酵素標識遊離ハプトグロビン抗体(モノクローナ
ル1−1 型抗体) 酵素標識遊離ハプトグロビン抗体(モノクローナル2−
1 型抗体) 酵素標識遊離ハプトグロビン抗体(モノクローナル2−
2 型抗体) (4)洗浄液 (5)酵素基質 (6)緩衝液 (7)反応停止液 (8)ハプトグロビン抗体感作ラテックス(モノクロー
ナル1−1 型抗体) ハプトグロビン抗体感作ラテックス(モノクローナル2
−1 型抗体) ハプトグロビン抗体感作ラテックス(モノクローナル2
−2 型抗体) B.測定操作 (1)スライドグラスの3ヶ所に検体各30ulを取
り、3種類のハプトグロビン感作ラテックス30ulを
添加、混和し、室温で5分間インキュベートし、凝集像
を観察し、ハプトグロビン型を決定した。 (2)小試験管に、検体1mlと遊離ヘモグロビンを固
定化したアガロースを封入したカートリッジ各1個を分
取、混和し、室温で30分間インキュベートした。 (3)各試験管の内容液を捨て、試験管内の遊離ヘモグ
ロビンを固定化したアガロースを封入したカートリッジ
を洗浄液2mlで2回洗浄、水切りをした。 (4)(3)の操作をした各試験管に、既知の型の酵素
標識遊離ハプトグロビン抗体(モノクローナル1−1型
抗体または、モノクローナル2−1型抗体、または、モ
ノクローナル2−2型抗体)2mlを添加、混和し、3
7℃で30分間インキュベートした。 (5)(4)の操作をした各試験管の内容液を捨て、試
験管内の遊離ヘモグロビンを固定化したアガロースを封
入したカートリッジを洗浄液2mlで3回洗浄、水切り
をした。 (6)(5)の操作をした各試験管に酵素基質液500
ulを添加、混和し、37℃で30分間インキュベート
した。 (7)(6)の操作をした各試験管に反応停止液2ml
を添加、混和した。 (8)ブランクを対照にして、492nmの吸光度を測
定した。 (9)標準遊離ハプトグロビンを用いて同一操作をして
描いた3種類(1−1型、2−1型、2−2型)の検量
線から検体の遊離ハプトグロビン量を求めた。Embodiment 5. A. Composition of Free Haptoglobin Quantification Kit (1) Cartridge encapsulating agarose with immobilized free hemoglobin (2) Standard free haptoglobin (3) Enzyme-labeled free haptoglobin antibody (monoclonal type 1-1 antibody) Enzyme-labeled free haptoglobin antibody (monoclonal 2) −
Type 1 antibody) Enzyme-labeled free haptoglobin antibody (monoclonal 2-
Type 2 antibody) (4) washing solution (5) enzyme substrate (6) buffer solution (7) reaction stop solution (8) haptoglobin antibody-sensitized latex (monoclonal type 1-1 antibody) haptoglobin antibody-sensitized latex (monoclonal 2)
-1 type antibody) Haptoglobin antibody-sensitized latex (monoclonal 2
-2 type antibody) B. Measurement operation (1) 30 ul of each sample was taken in 3 places on a slide glass, 30 ul of 3 types of haptoglobin-sensitized latex were added and mixed, and the mixture was incubated at room temperature for 5 minutes, and an agglutination image was observed to determine the haptoglobin type. (2) 1 ml of each of the cartridges containing 1 ml of the sample and agarose on which free hemoglobin was immobilized were collected in a small test tube, mixed, and incubated at room temperature for 30 minutes. (3) The content liquid in each test tube was discarded, and the cartridge containing agarose on which free hemoglobin was immobilized was washed twice with 2 ml of the washing solution and drained. (4) 2 ml of a known type of enzyme-labeled free haptoglobin antibody (monoclonal type 1-1 antibody, monoclonal 2-1 type antibody, or monoclonal 2-2 type antibody) in each test tube subjected to the operation of (3) Add and mix 3
Incubated at 7 ° C for 30 minutes. (5) The liquid content of each test tube operated in (4) was discarded, and the cartridge containing agarose in which free hemoglobin was immobilized was washed 3 times with 2 ml of the washing liquid and drained. (6) The enzyme substrate solution 500 is added to each of the test tubes subjected to the operations of (5).
ul was added, mixed, and incubated at 37 ° C. for 30 minutes. (7) 2 ml of reaction stop solution in each test tube operated in (6)
Was added and mixed. (8) Using the blank as a control, the absorbance at 492 nm was measured. (9) The amount of free haptoglobin of the sample was determined from the calibration curves of three types (1-1 type, 2-1 type, 2-2 type) drawn by performing the same operation using standard free haptoglobin.
【0011】[0011]
【発明の効果】本発明は、遊離ヘモグロビンを固定化し
た実験器具、または遊離ヘモグロビンを固定化したアフ
ィニティークロマトグラフィー担体を封入したカートリ
ッジを用いることにより、従来、検査方法が存在しなか
った遊離ハプトグロビンを定量する為に考案された。本
発明によれば、 (1)従来、不可能であった遊離ハプトグロビンの定量
が可能となる。 (2)従来法(非特異吸着法)に比較して、遊離ヘモグ
ロビンを固定化したアフィニティークロマトグラフィー
担体を封入したカートリッジは、検体を多量に吸着する
ことが可能となり、検体の希釈が不要になる。 (3)従来、定量が不能であった遊離ハブトグロビンの
定量が可能となる。 (4)EIA法(サンドイッチ法)と比較して、抗体使
用量が半減する。 という利点がある。INDUSTRIAL APPLICABILITY The present invention uses a laboratory instrument having free hemoglobin immobilized thereon, or a cartridge containing an affinity chromatography carrier having free hemoglobin immobilized therein, to eliminate free haptoglobin which has not been present in the conventional test method. It was devised to quantify. According to the present invention, (1) it becomes possible to quantify free haptoglobin, which has been impossible in the past. (2) Compared with the conventional method (non-specific adsorption method), the cartridge containing the affinity chromatography carrier on which free hemoglobin is immobilized can adsorb a large amount of the sample and does not require dilution of the sample. .. (3) It becomes possible to quantify free habutoglobin, which was conventionally impossible to quantify. (4) The amount of antibody used is halved as compared with the EIA method (sandwich method). There is an advantage that.
Claims (1)
したカートリッジを用いたハプトグロビンの定量方法。 【請求項2】−N=CH−(CH2)n−CH=N−、
−CONH−等の反応性官能基を固相化し、その官能基
に、遊離ヘモグロビンを共有結合させた試験管、ビー
ズ、マイクロタイタープレート、磁性粒子等の実験器具
を用いたハプトグロビンの定量方法。 【請求項3】「請求項1」、「請求項2」、「請求項
3」の実施の為に構成された遊離ハプトグロビン定量キ
ット。 【請求項4】「請求項1」、「請求項2」、「請求項
3」に記載された遊離ヘモグロビンを共有結合させた実
験器具。 【請求項5】ハプトダロビン抗体(モノクローナル1−
1型抗体、モノクローナル2−1型抗体、モノクローナ
ル2−2型抗体)を感作したラテックスまたは、赤血球
を用いた凝集法によるハプトグロビン型の判定方法。What is claimed is: 1. A method for quantifying haptoglobin using a cartridge in which a carrier on which free hemoglobin is immobilized is enclosed. Wherein -N = CH- (CH 2) n -CH = N-,
A method for quantifying haptoglobin using a test tube, beads, microtiter plate, magnetic particles, or other experimental device in which a reactive functional group such as -CONH- is immobilized on a solid phase and free hemoglobin is covalently bonded to the functional group. 3. A free haptoglobin quantification kit configured for carrying out "claim 1", "claim 2" and "claim 3". 4. An experimental instrument to which the free hemoglobin according to claim 1, claim 2 or claim 3 is covalently bound. 5. A haptodalobin antibody (monoclonal 1-
Haptoglobin type determination method by agglutination method using latex sensitized with type 1 antibody, monoclonal 2-1 type antibody, monoclonal 2-2 type antibody) or red blood cells.
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP18207691A JPH055738A (en) | 1991-04-20 | 1991-04-20 | Determining method of free haptoglobin |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP18207691A JPH055738A (en) | 1991-04-20 | 1991-04-20 | Determining method of free haptoglobin |
Publications (1)
Publication Number | Publication Date |
---|---|
JPH055738A true JPH055738A (en) | 1993-01-14 |
Family
ID=16111932
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
JP18207691A Pending JPH055738A (en) | 1991-04-20 | 1991-04-20 | Determining method of free haptoglobin |
Country Status (1)
Country | Link |
---|---|
JP (1) | JPH055738A (en) |
Cited By (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO1999024833A1 (en) * | 1997-11-12 | 1999-05-20 | The University Court Of The University Of Glasgow | Haptoglobin assay |
JP2003511701A (en) * | 1999-10-12 | 2003-03-25 | ザ ユニバーシティー コート オブ ザ ユニバーシティー オブ グラスゴウ | Assay method for mastitis to detect haptoglobin in milk |
-
1991
- 1991-04-20 JP JP18207691A patent/JPH055738A/en active Pending
Cited By (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO1999024833A1 (en) * | 1997-11-12 | 1999-05-20 | The University Court Of The University Of Glasgow | Haptoglobin assay |
US6451550B1 (en) | 1997-11-12 | 2002-09-17 | The University Court Of The University Of Glasgow | Haptoglobin assay |
JP2003511701A (en) * | 1999-10-12 | 2003-03-25 | ザ ユニバーシティー コート オブ ザ ユニバーシティー オブ グラスゴウ | Assay method for mastitis to detect haptoglobin in milk |
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