JPH0677019B2 - Enzyme-labeled antibody stabilization method - Google Patents
Enzyme-labeled antibody stabilization methodInfo
- Publication number
- JPH0677019B2 JPH0677019B2 JP59004855A JP485584A JPH0677019B2 JP H0677019 B2 JPH0677019 B2 JP H0677019B2 JP 59004855 A JP59004855 A JP 59004855A JP 485584 A JP485584 A JP 485584A JP H0677019 B2 JPH0677019 B2 JP H0677019B2
- Authority
- JP
- Japan
- Prior art keywords
- enzyme
- labeled antibody
- antibody
- afp
- peroxidase
- 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 - Lifetime
Links
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
- G01N33/531—Production of immunochemical test materials
- G01N33/532—Production of labelled immunochemicals
- G01N33/535—Production of labelled immunochemicals with enzyme label or co-enzymes, co-factors, enzyme inhibitors or enzyme substrates
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)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Description
【発明の詳細な説明】 本発明は酵素標識抗体の安定化法に関し、更に詳しくは
酵素標識抗体を凍結乾燥品などの製剤状態で、長期間に
亘って安定化することができる方法に関する。The present invention relates to a method for stabilizing an enzyme-labeled antibody, and more particularly to a method capable of stabilizing an enzyme-labeled antibody in a formulation such as a lyophilized product for a long period of time.
従来より免疫学的手法を用いて抗原抗体反応によって生
体液中の様々な物質を測定することが行なわれてきた。
この様な手法には、例えば毛細管沈降法、免疫比濁法、
ネフェロメリックイムノアッセイ、ラテックス凝集法、
ラジオイムノアッセイ等の測定法が利用されている。と
ころが、これらの測定法は、測定感度が適当でなかった
り、操作が煩雑であったり、放射性物質の処理等に問題
があるなど、通常の臨床検査において不都合を来してい
る。そこで、近年になってエンザイムイムノアッセイの
如く抗原又は抗体に酵素を標識し、これによって微量成
分を正確に測定する方法が開発され盛んに利用されてい
る。また、特願昭57−3314号明細書には、酵素を標識し
た抗原あるいは抗体を用いて抗原抗体反応を行なわせ、
会合の結果生ずる酵素活性の変化を光学的に測定し、目
的の抗原あるいは抗体を定量する均一系の分析方法が記
載されており、非常に簡便且つ正確な方法であるため、
日常的な臨床検査への利用が期待されている。ところ
が、酵素を標識した抗体は、粉末及び凍結乾燥品などの
製剤としての長期の安定性に問題があり、実用の妨げと
なっていることから、長期保存状態においても酵素活性
が低下する様なことのない酵素標識抗体を開発すること
が強く望まれていた。Conventionally, various substances in a biological fluid have been measured by an antigen-antibody reaction using an immunological method.
Such methods include, for example, capillary sedimentation method, immunoturbidimetric method,
Nepherometric immunoassay, latex agglutination method,
Measurement methods such as radioimmunoassay are used. However, these measuring methods are inconvenient in normal clinical examinations, such as inadequate measuring sensitivity, complicated operation, and problems with treatment of radioactive substances. Therefore, in recent years, a method of labeling an antigen or an antibody with an enzyme, such as an enzyme immunoassay, and thereby accurately measuring a trace amount of the component has been developed and actively used. Further, Japanese Patent Application No. 57-3314 discloses that an antigen-antibody reaction is carried out using an enzyme-labeled antigen or antibody,
Optically measuring the change in enzyme activity resulting from the association, a homogeneous analysis method for quantifying the antigen or antibody of interest has been described, and is a very simple and accurate method.
It is expected to be used for daily clinical tests. However, an enzyme-labeled antibody has a problem in long-term stability as a preparation such as a powder and a lyophilized product, which impedes its practical use. It has been strongly desired to develop a unique enzyme-labeled antibody.
本発明は、この様な実情に鑑みてなされたものであり、
その目的とするところは、製造時から使用時に亘る長期
保存状態においても酵素活性の低下が極めて少ないもの
となる、酵素標識抗体の安定化法を提供することにあ
る。The present invention has been made in view of such circumstances.
It is an object of the invention to provide a method for stabilizing an enzyme-labeled antibody, which has a very small decrease in enzyme activity even in a long-term storage state from the production to the use.
即ち、本発明はペルオキシダーゼ標識抗体を含む凍結乾
燥品において、該ペルオキシダーゼ標識抗体を安定化す
るために、シュークロースおよびEDTAを含有させること
を特徴とするものである。又、本発明はペルオキシダー
ゼ標識抗体を含む凍結乾燥品において、該ペルオキシダ
ーゼ標識抗体を安定化するために、シュークロースおよ
びフィコール400（商品名、ファルマシア・ファインケ
ミカル社製）を含有させることを特徴とするものでもあ
る。That is, the present invention is characterized in that a lyophilized product containing a peroxidase-labeled antibody contains sucrose and EDTA in order to stabilize the peroxidase-labeled antibody. The present invention is also characterized in that, in a lyophilized product containing a peroxidase-labeled antibody, sucrose and Ficoll 400 (trade name, manufactured by Pharmacia Fine Chemical Co., Ltd.) are contained in order to stabilize the peroxidase-labeled antibody. But also.
本発明に使用する酵素標識抗体は、抗α−フェトプロテ
イン（AFP）、抗IgE、抗Ｄ−フルクトース−1,6−二燐
酸（抗FDP）、抗フェリチン、抗カ−シノ−マエムブリ
オニックアンチゲン、抗β２マイクログロブリン等の抗
体をペルオキシダーゼ、アルカリ性フォスファターゼ、
β−ガラクトシダーゼ、グルコアミラーゼ、アセチルコ
リンエステラーゼ、マラテデヒドロゲナーゼ等の酵素で
標識したものである。The enzyme-labeled antibody used in the present invention includes anti-α-fetoprotein (AFP), anti-IgE, anti-D-fructose-1,6-diphosphate (anti-FDP), anti-ferritin, anti-carcino-maembrionic anti-antibody. Gen, anti-β 2 microglobulin antibody, peroxidase, alkaline phosphatase,
It is labeled with an enzyme such as β-galactosidase, glucoamylase, acetylcholinesterase, and malate dehydrogenase.
酵素標識抗体の調製には公知の試薬が使用できる。たと
えば、グルタルアルデヒド、カルボジイミド、ビスマレ
イミド、２個の異なる官能基を有する試薬等があり、ま
た、ペルオキシダーゼの糖鎖を過沃素酸で酸化してアル
デヒド基にする方法も有効である。これらの試薬を用い
抗体の反応性を保った状態で酵素を標識する。Known reagents can be used to prepare the enzyme-labeled antibody. For example, there are glutaraldehyde, carbodiimide, bismaleimide, reagents having two different functional groups, and the like, and a method of oxidizing a sugar chain of peroxidase with periodic acid to an aldehyde group is also effective. The enzyme is labeled with these reagents while maintaining the reactivity of the antibody.
本発明で使用する酵素標識抗体として特に好ましくは、
マレイミド導入ペルオキシダーゼとＦ（ab′）２とが結
合した酵素標識抗体である。抗体Ｆ（ab′）２画分の調
製は、公知の方法により、例えば抗FDP、抗IgE等をペプ
シン消化するなどして得られる。Particularly preferably as the enzyme-labeled antibody used in the present invention,
This is an enzyme-labeled antibody in which maleimide-introduced peroxidase and F (ab ′) 2 are bound. The antibody F (ab ') 2 fraction can be prepared by a known method, for example, by digesting anti-FDP, anti-IgE, etc. with pepsin.
酵素標識抗体は、通常、リン酸緩衝液、トリス緩衝液等
のpH6.5〜8.0の適宜の緩衝液と混合され液状（以下、製
剤原液という）にした後、常法に従って、凍結乾燥、噴
霧乾燥などにより粉粒状製剤とされる。この場合、本発
明で使用するシュークロースおよびEDTAあるいはシュー
クロースおよびフィコール400（商品名、ファルマシア
・ファインケミカル社製）は、前記緩衝液との混合時に
酵素標識抗体に添加されるのが好適である。なお、使用
する前記緩衝液はリン酸緩衝液又はトリス緩衝液である
ことが好ましいが、これらに限定されない。The enzyme-labeled antibody is usually mixed with an appropriate buffer solution having a pH of 6.5 to 8.0 such as a phosphate buffer solution or a Tris buffer solution to give a liquid (hereinafter, referred to as a stock solution of the formulation), and then freeze-dried and sprayed according to a conventional method. It is made into a powdery and granular preparation by drying or the like. In this case, sucrose and EDTA or sucrose and Ficoll 400 (trade name, manufactured by Pharmacia Fine Chemical Co.) used in the present invention are preferably added to the enzyme-labeled antibody when mixed with the buffer solution. The buffer solution used is preferably a phosphate buffer solution or a Tris buffer solution, but is not limited thereto.
本発明で使用する二糖類以上の非還元糖類及び糖アルコ
ールとしては、シュークロース、トレハロース、ツラノ
ース、ラフィノース、メチレトース等の少糖類、デキス
トラン等の多糖類などの糖類、フィコール400（商品
名、ファルマシア・ファインケミカル社製；シュクロー
スと、エピクロルヒドリンとの分岐を有する反応生成
物；分子量40万）糖の等誘導体、及びイノシトール、マ
ンニトール、ソルビトール等の糖アルコールが挙げられ
る。これらの糖類及び糖アルコールの使用量は、製剤原
液に対して、一般に、１重量％以上で十分な効果が得ら
れる。また、シュクロースを例にとれば、2.5重量％以
上の添加で更に良好な結果が得られる。As non-reducing sugars and sugar alcohols of disaccharides or more used in the present invention, sucrose, trehalose, turanose, raffinose, oligosaccharides such as methyletose, saccharides such as polysaccharides such as dextran, Ficoll 400 (trade name, Pharmacia. Fine Chemical Co .; a reaction product having a branch of sucrose and epichlorohydrin; a molecular weight of 400,000) a derivative of sugar or the like, and a sugar alcohol such as inositol, mannitol or sorbitol. The amount of these sugars and sugar alcohols used is generally 1% by weight or more with respect to the stock solution of the preparation, and a sufficient effect is obtained. In addition, taking sucrose as an example, even better results can be obtained by adding 2.5% by weight or more.
本発明によれば、酵素標識抗体製剤の安定化に極めて甚
大な効果が奏されるが、とりわけ凍結乾燥品を得るなど
製剤時の安定化、ならびに、凍結乾燥品などの製剤とし
ての長期に亘る安定化に良好なる結果をもたらす。According to the present invention, an extremely enormous effect is exerted on the stabilization of an enzyme-labeled antibody preparation, but especially during preparation such as obtaining a lyophilized product, and for a long time as a preparation such as a lyophilized product. It gives good stabilization results.
以下に示す試験例により、本発明を更に詳しく説明す
る。The present invention will be described in more detail by the following test examples.
試験例１ α−フェトプロテイン（AFP）の測定におけ
る各種安定化剤の効果 特願昭57−3314号明細書に記載された方法に基き、酵素
標識抗体の調製、並びにAFPの測定を行なった。Test Example 1 Effects of various stabilizers on the measurement of α-fetoprotein (AFP) Based on the method described in Japanese Patent Application No. 57-3314, enzyme-labeled antibodies were prepared and AFP was measured.
ａ）抗AFP抗体Ｆ（ab′）２画分の調製： 西等の方法（Cancer Res.,30,2507〜2513,1970）により
臍帯血清より抽出し精製したAFPをフロイントの完全ア
ジュバントと等量混合し、ウサギに免疫して抗AFPウサ
ギ血清を得た。この抗血清よりエベレイ等の方法（J.So
lidphase Biochem.,2,45〜78,1977）によって抗体を精
製した。0.1M酢酸緩衝液（pH4.5）に対して透析した抗
体に２％重量のペプシン（ベーリンガー社製品）を加え
37℃において48時間消化した後、セファデックスＧ−20
0カラムを用いて抗AFP抗体Ｆ（ab′）２画分を得た。a) Preparation of anti-AFP antibody F (ab ′) 2 fraction: AFP extracted and purified from umbilical cord serum by the method of Nishi et al. (Cancer Res., 30, 2507 to 2513, 1970) and equivalent to Freund's complete adjuvant After mixing, the rabbit was immunized to obtain anti-AFP rabbit serum. From this antiserum, the method of Everey et al.
The antibody was purified by lidphase Biochem., 2, 45-78, 1977). To the antibody dialyzed against 0.1M acetate buffer (pH 4.5), add 2% by weight of pepsin (product of Boehringer).
After digesting at 37 ℃ for 48 hours, Sephadex G-20
An anti-AFP antibody F (ab ') 2 fraction was obtained using a 0 column.
ｂ）マレイミド基導入HRPの調製： ナカネ等の方法（The J. of Histochem.＆ Cytochem.,2
2−12,1084〜1091,1974）に準じ、西洋ワサビペルオキ
シダーゼ（HRP）9mgとテトラメチレンジアミン15mgを結
合させ、アミノ基を導入したHRPを得た。このアミノ基
導入HRP2.3mgに北川らの方法（臨床化学,6巻,3号,178−
186,1978）によりメタ−マレイミドベンゾイル−Ｎ−ヒ
ドロキシスクシンイミドエステル（ピアスケミカル社製
品）1.8mgを加え30℃において30分間反応させた後、セ
ファデックスＧ−25カラムを用いて分画しマレイミド基
を導入したHRPを得た。b) Preparation of maleimide group-introduced HRP: Nakane et al. (The J. of Histochem. & Cytochem., 2
2-12,1084-1091,1974), 9 mg of horseradish peroxidase (HRP) and 15 mg of tetramethylenediamine were combined to obtain HRP having an amino group introduced. To this amino group-introduced HRP 2.3 mg, the method of Kitagawa et al. (Clinical Chemistry, Volume 6, Issue 3, 178-
186, 1978) 1.8 mg of meta-maleimidobenzoyl-N-hydroxysuccinimide ester (produced by Pierce Chemical Co., Ltd.) was added and reacted at 30 ° C. for 30 minutes, then fractionated using a Sephadex G-25 column to remove maleimide groups. The introduced HRP was obtained.
ｃ）酵素標識抗体の調製： ａ）で調製した抗体Ｆ（ab′）２に最終濃度12.5mMの２
−メルカプトエチルアミンを加え90分間反応後セファデ
ックスＧ−25カラムを用いて分画して得られた抗体Fa
b′6mgに実施例２−ｂ）で調製したマレイミド基導入HR
P1.5mgを加え37℃において30分間反応後、室温で一夜静
置した。これをセファデックスＧ−200カラムを用いて
分画し酵素標識抗体を得た。c) Preparation of enzyme-labeled antibody: The antibody F (ab ′) 2 prepared in a) was added with 2 at a final concentration of 12.5 mM.
-An antibody Fa obtained by adding mercaptoethylamine for 90 minutes and fractionating using a Sephadex G-25 column
b′6 mg with maleimide group-introduced HR prepared in Example 2-b)
After adding P1.5 mg and reacting at 37 ° C. for 30 minutes, the mixture was allowed to stand at room temperature overnight. This was fractionated using a Sephadex G-200 column to obtain an enzyme-labeled antibody.
ｄ）AFPの測定： リン酸緩衝液（pH7.0）で調製したAFP希釈系列より0.05
ml、６％ポリエチレングリコール＃6000のリン酸緩衝溶
液より0.05ml、並びにｃ）の酵素標識抗体0.05mlをそれ
ぞれ試験管にとり、37℃において20分間反応させた後、
0.75mM4−アミノアンチピリン、25mMフェノール10mM過
酸化水素からなる基質呈色液0.5mlを加え37℃10分間反
応後、反応停止液2.0mlを加え反応を停止した後、波長5
00nmにおける吸光度を測定する。d) Measurement of AFP: 0.05 from AFP dilution series prepared with phosphate buffer (pH 7.0)
ml, 0.05 ml of a 6% polyethylene glycol # 6000 phosphate buffer solution, and 0.05 ml of the enzyme-labeled antibody of c) were placed in test tubes and reacted at 37 ° C. for 20 minutes.
After adding 0.5 ml of a substrate coloring solution consisting of 0.75 mM 4-aminoantipyrine and 25 mM phenol 10 mM hydrogen peroxide and reacting at 37 ° C for 10 minutes, 2.0 ml of a reaction stop solution was added to stop the reaction, and then a wavelength of 5
Measure the absorbance at 00 nm.
上記測定法を用いて以下の如く調製した凍結乾燥品の安
定性を調べた。The stability of the freeze-dried product prepared as described below was examined using the above-mentioned measurement method.
403nmの吸光度0.025となるAFP.酵素標識抗体に表−１に
示した0.5〜５％の各種安定化剤と2.5％牛血清アルブミ
ンと20mMの各種緩衝液を含有する原液を調製しその1ml
を10mlバイアル瓶に分注し凍結乾燥を行った。An AFP enzyme-labeled antibody with an absorbance of 403 nm of 0.025 was prepared as a stock solution containing 0.5 to 5% of various stabilizers shown in Table-1, 2.5% bovine serum albumin and 20 mM of various buffer solutions, and 1 ml thereof was prepared.
Was dispensed into a 10 ml vial and freeze-dried.
AFP測定時には生理食塩水2.5mlにて１バイアルを溶解し
その0.02mlを上記測定時に用いた。At the time of AFP measurement, one vial was dissolved with 2.5 ml of physiological saline and 0.02 ml thereof was used for the above measurement.
表−１に各種条件での凍結乾燥直後と37℃にて１ケ月間
保存した酵素標識抗体のAFP測定時のAFP希釈系列の吸光
度を各々、原液、凍結乾燥直後を100％として残存活性
（％）であらわした。Table 1 shows the residual activity (%) of the absorbance of the AFP dilution series immediately after freeze-drying under various conditions and at the time of AFP measurement of the enzyme-labeled antibody stored at 37 ° C for 1 month as 100% for the stock solution and immediately after freeze-drying, respectively. ).
安定剤を添加した場合においては凍結乾燥時の低下及び
長期保存に対して著明な効果が見受けられた。When a stabilizer was added, a remarkable effect was observed on the decrease during freeze-drying and long-term storage.
一般的に安定剤としてよく用いられる牛血清アルブミン
による安定化（No.2とNo.9を対比）は弱く本発明の安定
化方法では極めて良好に安定化されている。またトリス
緩衝液とリン酸緩衝液とによる差は見られない。Stabilization (both No. 2 and No. 9) by bovine serum albumin, which is commonly used as a stabilizer, is weak, and it is extremely well stabilized by the stabilization method of the present invention. Further, no difference between the Tris buffer solution and the phosphate buffer solution is observed.
試験例2.pH及び二種混合による影響 試験例１の酵素標識抗体を用い各種化合物、及び安定剤
を添加し、20mMの各種緩衝液にてpHを変化させた原液を
調製しその1mを10mlバイアル瓶に分注し凍結乾燥を行っ
た。Test Example 2. Effect of pH and mixing of two kinds Using the enzyme-labeled antibody of Test Example 1, various compounds and a stabilizer were added, and a stock solution was prepared by changing the pH with various buffers of 20 mM. It was dispensed into a vial and freeze-dried.
37℃１ケ月間の保存安定性を試験例1.と同様にAFP測定
時のAFP希釈系列の吸光度を凍結乾燥直後を100％として
残存活性（％）で表−２に示した。The storage stability at 37 ° C. for one month was shown in Table 2 as the residual activity (%) with the absorbance of the AFP dilution series at the time of AFP measurement as 100% immediately after lyophilization, as in Test Example 1.
pH変化による影響は極めて少く、安定化剤の二種混合に
おいても極めて良好な保存性が得られた。還元糖との組
合せでは還元糖の影響を受け若干安定性が不良であっ
た。安定化剤無添加の場合では残存活性は殆ど無くなっ
ている。The effect of pH change was extremely small, and very good storage stability was obtained even when two kinds of stabilizers were mixed. In the combination with reducing sugar, the stability was slightly poor due to the influence of reducing sugar. When the stabilizer is not added, the residual activity is almost lost.
試験例3.安定化剤の濃度と安定性 試験例１の酵素標識抗体を用い安定化剤の濃度を変化さ
せた20mMトリス緩衝液pH7.5を含む原液を調製しその1ml
を10mlバイアル瓶に分注し凍結乾燥を行った。 Test Example 3 Stabilizer Concentration and Stability Using the enzyme-labeled antibody of Test Example 1 to prepare a stock solution containing 20 mM Tris buffer pH 7.5 in which the concentration of the stabilizer was changed, 1 ml thereof was prepared.
Was dispensed into a 10 ml vial and freeze-dried.
図−１に37℃１ケ月間の残存活性（％）（AFP濃度800ng
/mlで曲線１、100ng/mlで曲線２）を試験例１と同様にA
FP測定時のAFP希釈系列の吸光度を凍結乾燥直後の吸光
度を100％として表した。安定化剤として好ましくは2.5
％以上あれば良いことが判る。Figure 1 shows the residual activity (%) for 1 month at 37 ℃ (AFP concentration 800 ng)
curve 1 at 100 ng / ml, curve 2 at 100 ng / ml) as in Test Example 1
The absorbance of the AFP dilution series at the time of FP measurement was expressed with the absorbance immediately after freeze-drying as 100%. As a stabilizer preferably 2.5
It turns out that it is good if it is at least%.
図面は、本発明方法により安定化した酵素標識抗体の安
定化剤濃度による活性安定化の変化を示した図である。 曲線１……AFP濃度800ng/ml 曲線２……AFP濃度100ng/mlThe drawings are diagrams showing changes in activity stabilization of enzyme-labeled antibodies stabilized by the method of the present invention depending on the concentration of the stabilizer. Curve 1 …… AFP concentration 800ng / ml Curve 2 …… AFP concentration 100ng / ml
Claims (2)
品において、シュークロースおよびEDTAを含有させるこ
とを特徴とするペルオキシダーゼ標識抗体の安定化法。1. A method for stabilizing a peroxidase-labeled antibody, which comprises adding sucrose and EDTA to a lyophilized product containing the peroxidase-labeled antibody.
品において、シュークロースおよびフィコール400（商
品名、ファルマシア・ファインケミカル社製）を含有さ
せることを特徴とするペルオキシダーゼ標識抗体の安定
化法。2. A method for stabilizing a peroxidase-labeled antibody, which comprises adding sucrose and Ficoll 400 (trade name, manufactured by Pharmacia Fine Chemical Co., Ltd.) to a freeze-dried product containing the peroxidase-labeled antibody.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP59004855A JPH0677019B2 (en) | 1984-01-17 | 1984-01-17 | Enzyme-labeled antibody stabilization method |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP59004855A JPH0677019B2 (en) | 1984-01-17 | 1984-01-17 | Enzyme-labeled antibody stabilization method |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| JPS60149972A JPS60149972A (en) | 1985-08-07 |
| JPH0677019B2 true JPH0677019B2 (en) | 1994-09-28 |
Family
ID=11595290
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP59004855A Expired - Lifetime JPH0677019B2 (en) | 1984-01-17 | 1984-01-17 | Enzyme-labeled antibody stabilization method |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPH0677019B2 (en) |
Families Citing this family (12)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| ATE68524T1 (en) * | 1985-07-09 | 1991-11-15 | Quadrant Bioresources Ltd | PROTECTION OF PROTEINS AND THE LIKE. |
| JPS63131065A (en) * | 1986-11-20 | 1988-06-03 | Yatoron:Kk | Refining of antibody, measurement of isozyme and reagent |
| GB8716826D0 (en) * | 1987-07-16 | 1987-08-19 | Tills D | Protection of proteinaceous reagents |
| WO1989009402A1 (en) * | 1988-03-30 | 1989-10-05 | Toray Industries, Inc. | FREEZE-DRIED COMPOSITION CONTAINING ENZYME-LABELED ANTIHUMAN INTERFERON-beta ANTIBODY AND ENZYMATIC IMMUNOASSAY KIT CONTAINING THE COMPOSITION |
| US5102788A (en) * | 1988-11-21 | 1992-04-07 | Hygeia Sciences, Inc. | Immunoassay including lyophilized reactant mixture |
| JP3659985B2 (en) * | 1993-12-28 | 2005-06-15 | 三栄源エフ・エフ・アイ株式会社 | Trehalase and method for producing saccharides using the enzyme |
| TW426737B (en) * | 1994-06-27 | 2001-03-21 | Hayashibara Biochem Lab | Saccharide composition with reduced reducibility, and preparation and uses thereof |
| TW466116B (en) | 1997-03-04 | 2001-12-01 | Hayashibara Biochem Lab | Reduction inhibitory agent for active-oxygen eliminating activity, method for inhibiting the reduction of said activity, and composition containing said agent |
| KR20110091822A (en) | 2002-02-14 | 2011-08-12 | 추가이 세이야쿠 가부시키가이샤 | Antibody-containing solution pharmaceuticals |
| JP7269906B2 (en) * | 2019-10-29 | 2023-05-09 | 三洋化成工業株式会社 | Immunoassay reagent, immunoassay kit, and immunoassay method |
| CN116298317A (en) * | 2023-03-14 | 2023-06-23 | 浙江夸克生物科技有限公司 | Alpha fetoprotein determination kit based on latex immunoturbidimetry |
| CN116466092A (en) * | 2023-03-21 | 2023-07-21 | 浙江夸克生物科技有限公司 | Kit for quantitatively determining uroretinol binding protein |
Family Cites Families (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS57118790A (en) * | 1981-01-14 | 1982-07-23 | Takeda Chem Ind Ltd | Freeze-dried substance containing beta-d-galactosidase |
| JPS58149700A (en) * | 1982-03-02 | 1983-09-06 | Takeda Chem Ind Ltd | Composite containing peroxidase, its preparation and reagent |
-
1984
- 1984-01-17 JP JP59004855A patent/JPH0677019B2/en not_active Expired - Lifetime
Also Published As
| Publication number | Publication date |
|---|---|
| JPS60149972A (en) | 1985-08-07 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CA1206877A (en) | Immunoassay for nonenzymatically glucosylated proteins and protein fragments - an index of glycemia | |
| US4863875A (en) | Dye labelled antibodies as reagents for use in immunoassay systems | |
| US4959306A (en) | Labeling design for a binding assay reagent | |
| JPH0677019B2 (en) | Enzyme-labeled antibody stabilization method | |
| EP0840895B1 (en) | Methods of obtaining receptor:release ligand (reland) complexes and test assays | |
| JPS63292061A (en) | Selective immunological measuring method of complete flaw-free pracollagen peptide (iii type) and procollagen (iii type) | |
| EP0166583B1 (en) | Compositions for enzyme immunoassay of prostaglandins | |
| US4966838A (en) | Process for the immunological quantitative determination of T3 and/or T4 thyroid hormones, using thyroglobulin | |
| US5130234A (en) | Method of quantitative determination utilizing enzyme aggregation | |
| US4458013A (en) | Method for the immunoassay of elastase-1 and a set of reagents for such immunoassay | |
| JPS6223823B2 (en) | ||
| JP2968910B2 (en) | Antibodies against 1α, 25 (OH) 2 vitamin D3 and uses thereof | |
| EP0132292A2 (en) | Method of measuring biological ligands | |
| EP0327312B1 (en) | Receptor preincubation enzyme assay | |
| JP2866151B2 (en) | Method and kit for detecting calpastatin abnormality | |
| EP0079962B1 (en) | Immunoprecipitation assay | |
| JPS61241665A (en) | Stabilized sold phase reagent | |
| JPH06105252B2 (en) | Enzyme-linked immunosorbent assay for free thyroxine | |
| EP0184701B1 (en) | A method for determining a ligand | |
| JPH07103978A (en) | Measurement of free hemoglobin | |
| FR2596867A1 (en) | Immunological method of assaying erythropoietin | |
| CA1192490A (en) | Immunoprecipitation assay | |
| Hata et al. | Enzyme immunoassay of free triiodothyronine in serum. | |
| JPH11125635A (en) | Stabilized lyophilization product containing sensitized metal colloid and production thereof | |
| JPH0236353A (en) | Immunoassay |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| EXPY | Cancellation because of completion of term |