JPH01223349A - Reagent for judging anti d blood type - Google Patents
Reagent for judging anti d blood typeInfo
- Publication number
- JPH01223349A JPH01223349A JP5071088A JP5071088A JPH01223349A JP H01223349 A JPH01223349 A JP H01223349A JP 5071088 A JP5071088 A JP 5071088A JP 5071088 A JP5071088 A JP 5071088A JP H01223349 A JPH01223349 A JP H01223349A
- Authority
- JP
- Japan
- Prior art keywords
- reagent
- monoclonal antibody
- type
- agglutination
- blood type
- 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.)
- Granted
Links
- 239000003153 chemical reaction reagent Substances 0.000 title claims abstract description 35
- 239000008280 blood Substances 0.000 title claims description 40
- 210000004369 blood Anatomy 0.000 title claims description 31
- 108060003951 Immunoglobulin Proteins 0.000 claims abstract description 17
- 102000018358 immunoglobulin Human genes 0.000 claims abstract description 17
- 102000004169 proteins and genes Human genes 0.000 claims abstract description 8
- 108090000623 proteins and genes Proteins 0.000 claims abstract description 8
- 230000004520 agglutination Effects 0.000 abstract description 31
- 108091003079 Bovine Serum Albumin Proteins 0.000 abstract description 4
- PXIPVTKHYLBLMZ-UHFFFAOYSA-N Sodium azide Chemical compound [Na+].[N-]=[N+]=[N-] PXIPVTKHYLBLMZ-UHFFFAOYSA-N 0.000 abstract description 4
- QYSGYZVSCZSLHT-UHFFFAOYSA-N octafluoropropane Chemical compound FC(F)(F)C(F)(F)C(F)(F)F QYSGYZVSCZSLHT-UHFFFAOYSA-N 0.000 abstract description 4
- 239000000203 mixture Substances 0.000 abstract description 3
- 230000002421 anti-septic effect Effects 0.000 abstract 1
- 238000012360 testing method Methods 0.000 description 27
- 210000002966 serum Anatomy 0.000 description 16
- 210000000601 blood cell Anatomy 0.000 description 12
- 108010088751 Albumins Proteins 0.000 description 8
- 102000009027 Albumins Human genes 0.000 description 8
- 239000007788 liquid Substances 0.000 description 7
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 6
- 238000000034 method Methods 0.000 description 6
- 239000011780 sodium chloride Substances 0.000 description 6
- 238000004220 aggregation Methods 0.000 description 5
- 230000002776 aggregation Effects 0.000 description 5
- 239000000427 antigen Substances 0.000 description 4
- 102000036639 antigens Human genes 0.000 description 4
- 108091007433 antigens Proteins 0.000 description 4
- 238000001514 detection method Methods 0.000 description 4
- 238000004519 manufacturing process Methods 0.000 description 4
- 239000000725 suspension Substances 0.000 description 4
- 239000006285 cell suspension Substances 0.000 description 3
- 238000010790 dilution Methods 0.000 description 3
- 239000012895 dilution Substances 0.000 description 3
- 230000000694 effects Effects 0.000 description 3
- 238000005516 engineering process Methods 0.000 description 3
- 239000002504 physiological saline solution Substances 0.000 description 3
- 238000011160 research Methods 0.000 description 3
- 210000004408 hybridoma Anatomy 0.000 description 2
- 230000001965 increasing effect Effects 0.000 description 2
- 238000005259 measurement Methods 0.000 description 2
- 238000004062 sedimentation Methods 0.000 description 2
- 239000000243 solution Substances 0.000 description 2
- 102000016550 Complement Factor H Human genes 0.000 description 1
- 108010053085 Complement Factor H Proteins 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 230000007910 cell fusion Effects 0.000 description 1
- 230000015271 coagulation Effects 0.000 description 1
- 238000005345 coagulation Methods 0.000 description 1
- 238000012790 confirmation Methods 0.000 description 1
- 230000002708 enhancing effect Effects 0.000 description 1
- 239000011521 glass Substances 0.000 description 1
- 230000003053 immunization Effects 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 230000035935 pregnancy Effects 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 239000003755 preservative agent Substances 0.000 description 1
- 230000002335 preservative effect Effects 0.000 description 1
- 238000009666 routine test Methods 0.000 description 1
Landscapes
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
Description
【発明の詳細な説明】
〔産業上の利用分野〕
本発明はＲｈ式血液型のＲｈｏ （Ｄ）因子の検出を行
うための抗り血液型判定用試薬に関する。DETAILED DESCRIPTION OF THE INVENTION [Industrial Field of Application] The present invention relates to a reagent for determining blood type for detecting the Rho (D) factor of Rh blood type.
Ｒｈ式血液型はＬａｎｄｓｔｅｉｎｅｒとＷｉｅｎｅｒ
により発見されて以来、不適合輸血や不適合妊娠の検査
としてＡＢＯ式血液型と並び重要視されてきた。現在Ｒ
ｈ因子（抗原）は４０数種類が見つけられているが、通
常Ｒｈ （＋）または（−）の表現がＤ因子の陽性、陰
性を意味するように、日常検査においてこのＲｈｏ （
Ｄ）因子の検出が最も多く行われている。Rh blood type is Landsteiner and Wiener
Since its discovery, it has been considered as important as the ABO blood type as a test for incompatible blood transfusions and incompatible pregnancies. Currently R
More than 40 types of factor H (antigen) have been discovered, and as the expression Rh (+) or (-) usually means positive or negative for factor D, this Rho (antigen) is commonly used in routine tests.
D) Factors are most commonly detected.
Ｒｈｏ（Ｄ）因子の検出には抗り血液型判定用試薬を用
いて行うことができる。七ころがＲｈ式血液型の検査で
はＡＢＯ式血液型の検査のようにオモテ検査とウラ検査
という確認の手段がない。Detection of the Rho (D) factor can be performed using a reagent for blood type determination. In the Rh blood type test, unlike the ABO blood type test, there is no front and back test for confirmation.
従って、Ｒｈ式血液型の検査を行うにあたっては操作に
留意すると共に、用いる試薬は特に力価や特異性につい
て十分に保証されていることが必要である。Therefore, when testing Rh blood type, it is necessary to pay attention to the operation and to ensure that the reagents used are sufficiently guaranteed, particularly in terms of potency and specificity.
従来より抗り血液型判定用試薬としては、ヒトまたはそ
の他の動物を免疫して得られた抗血清である抗Ｄポリク
ローナル抗体を用いた抗り血液型判定用血清がある。Conventional reagents for determining blood type include serum for determining blood type using an anti-D polyclonal antibody, which is an antiserum obtained by immunizing humans or other animals.
一方、最近では１９７５年にＫａｈｌｅｒとＭｉｌｓｔ
ｅｉｎらの研究に始まるモノクローン抗体産生ハイブリ
ドーマの作成技術、所謂細胞融合の技術による抗Ｄモノ
クローナル抗体も作成されている（例えば、特開昭５９
−１３２８８８、特開昭６Ｏ−１３６５９９）、このモ
ノクローナル抗体は、従来のボリフローナル抗体に比べ
て、単クーロンより産生されるため高い特異性を有し、
均質なものが永続的に得られるので、これを使用した抗
り血液型判定用試薬の製造供給が大いに期待されている
ところである。On the other hand, recently in 1975 Kahler and Milst
Anti-D monoclonal antibodies have also been created using the so-called cell fusion technology, which is a technology for creating monoclonal antibody-producing hybridomas that began with research by Ein et al.
-132888, JP-A-6O-136599), this monoclonal antibody has higher specificity than conventional vorifloral antibodies because it is produced from a single clone;
Since a homogeneous product can be obtained permanently, there are great expectations for the production and supply of a reagent for determining blood type using this method.
しかし、抗Ｄモノクローナル抗体は特異性は優れている
が、これをそのまま用いてＲｈｏ（Ｄ）因子の検出を肉
眼で凝集として観察する場合、実用上刃価の点に問題が
残っていた。However, although the anti-D monoclonal antibody has excellent specificity, when using this antibody as it is to observe the detection of Rho (D) factor with the naked eye as aggregation, there remains a problem in terms of the practical edge value.
通常、生理食塩水中の血球浮遊液（抗原）はマイナス側
に荷電しているため、血球間で互いに反発しあっている
。従って血球間は一定の距離を保っていて、それ以上近
づ（ことはない、一方抗体であるＩｇＧの抗原結合部位
の長さは血球間を架橋させるには至らず、ＩｇＧ抗体と
血球は反応していてもＩｇＧ抗体のみでは肉眼的に血球
を凝集させることはできない、このため抗Ｄポリクロー
ナル抗体を用いた場合には、この輯集反応を補うのに普
通アルブミンを添加している。その結果血球間の反発エ
ネルギーが押さえられ、Ｒｈｏ（Ｄ）因子との凝集反応
が増強されて実用上刃価が上がり、肉眼で充分に凝集を
観察することができる。Normally, the blood cell suspension (antigen) in physiological saline is negatively charged, so the blood cells repel each other. Therefore, blood cells maintain a certain distance and do not get closer than that. On the other hand, the length of the antigen-binding site of IgG, which is an antibody, is not enough to bridge blood cells, and IgG antibodies and blood cells react. However, IgG antibodies alone cannot visually aggregate blood cells, so when anti-D polyclonal antibodies are used, albumin is usually added to compensate for this aggregation reaction. The repulsive energy between blood cells is suppressed, the agglutination reaction with Rho (D) factor is enhanced, the blade value is increased in practical use, and agglutination can be sufficiently observed with the naked eye.
しかし、抗Ｄモノクローナル抗体を用いた場合には、こ
のような蛋白質の添加だけではＲｈｏ （Ｄ）因子との
凝集反応が十分には増強されず、肉眼での凝集の観察は
困難であり、厚生省で定める抗り血液型判定用血清の製
造基準に達することが出来ない、従って、抗Ｄモノクロ
ーナル抗体を用いた抗り血液型判定用試薬は未だ実用化
されていないのが実情である。However, when anti-D monoclonal antibodies are used, the agglutination reaction with Rho (D) factor is not sufficiently enhanced by the addition of such proteins alone, and it is difficult to observe agglutination with the naked eye. The actual situation is that a reagent for anti-blood group determination using an anti-D monoclonal antibody has not yet been put into practical use.
本発明の目的は、Ｒｈｏ（Ｄ）因子の検出にあたり抗Ｄ
モノクローナル抗体を用いても、その凝集反応が充分に
増強され、実用上刃価の高い抗り血液型判定用試薬を提
供することにある。The purpose of the present invention is to detect anti-D
The object of the present invention is to provide a reagent for blood type determination whose agglutination reaction is sufficiently enhanced even when a monoclonal antibody is used and which has a high resistance value in practical use.
本発明者らは、このような課題を解決し上述の目的を達
成するため鋭意研究を進めた結果、蛋白質の存在下に、
抗Ｄモノクローナル抗体としてイムノグロブリンサブク
ラスがＩｇＧタイプのものとイムノグロブリンサブクラ
スが１８Ｍタイプのものを併用させると、Ｒｈｏ（Ｄ）
因子との凝集反応が顕著に増強され、肉眼で十分に凝集
を観察することができることを見い出し、さらに研究を
重ねて本発明を完成するに至った。The present inventors have carried out intensive research to solve these problems and achieve the above objectives, and as a result, in the presence of proteins,
When an anti-D monoclonal antibody with an immunoglobulin subclass of IgG type and an immunoglobulin subclass of 18M type are used together, Rho(D)
It was discovered that the agglutination reaction with the factor was significantly enhanced and the agglutination could be sufficiently observed with the naked eye, and after further research, the present invention was completed.
すなわち、本発明はイムノグロブリンサブクラスがＩｇ
Ｇタイプの抗Ｄモノクローナル抗体、イムノグロブリン
サブクラスが１８Ｍタイプの抗Ｄモノクローナル抗体お
よび蛋白質よりなることを特徴とするＲｈｏ（Ｄ）因子
検出のための抗り血液型判定用試薬に関するものである
。That is, the present invention provides that the immunoglobulin subclass is Ig.
The present invention relates to a reagent for determining blood type for detecting Rho (D) factor, which is characterized by comprising an anti-D monoclonal antibody of G type, an anti-D monoclonal antibody of immunoglobulin subclass 18M type, and a protein.
本発明の抗り血液型判定用試薬に用いる抗Ｄモノクロー
ナル抗体としては、イムノグロブリンサブクラスがＩｇ
Ｇタイプの抗Ｄモノクローナル抗体とイムノグロブリン
サブクラスが１８Ｍタイプの抗Ｄモノクローナル抗体が
併用して使用される。The anti-D monoclonal antibody used in the anti-blood type determination reagent of the present invention has an immunoglobulin subclass of Ig.
A G-type anti-D monoclonal antibody and an 18M-type anti-D monoclonal antibody are used in combination.
とりわけ、そのうちでも特異性が安定している点から、
ヒト−ヒトハイブリドーマから産生されたヒト型抗Ｄモ
ノクローナル抗体が好ましい。Especially, since the specificity is stable among them,
Humanized anti-D monoclonal antibodies produced from human-human hybridomas are preferred.
ＩｇＧタイプと１８Ｍタイプとの配合割合は、本発明の
目的を達成しうるものであれば特に限定はないが、通常
それぞれの力価として１：４から４：１、好ましくは１
：２から２：１の割合となるように配合される。The blending ratio of the IgG type and the 18M type is not particularly limited as long as it can achieve the purpose of the present invention, but usually the potency of each is 1:4 to 4:1, preferably 1.
:2 to 2:1 ratio.
本発明の抗り血液型判定用試薬に用いる蛋白質としては
、本発明の目的を満足するものはすべて用いることがで
きる。具体的な蛋白質の例としては、アルブミンがあげ
られる。As the protein used in the reagent for determining blood type of the present invention, any protein that satisfies the purpose of the present invention can be used. A specific example of the protein is albumin.
その添加量は、−概に限定されないが、通常５〜３０％
、好ましくは１０〜２５％の濃度になるように添加され
る。The amount added is generally 5 to 30%, but not limited to
, preferably at a concentration of 10 to 25%.
本発明の抗り血液型判定用試薬は上述のように構成され
ているが、当該試薬は厚生省の定める抗り血液型判定用
血清の製造基準に適合するものである。この製造基準の
うち凝集反応に関しては、凝集力試験と凝集価試験があ
り、凝集力試験は定められた条件のもとて陽性検体を用
い、凝集開始までの時間（秒）および凝集塊の大きさが
ｌａｗｎ”に達するまでの時間（秒）を測定する。また
凝集価試験は定められた条件のもとで２倍連続希釈し、
陽性を示す最高希釈倍数から凝集価を求める。The reagent for determining a negative blood type of the present invention is constructed as described above, and the reagent complies with the manufacturing standards for serum for determining a negative blood type set by the Ministry of Health and Welfare. Regarding the agglutination reaction in this manufacturing standard, there are an agglutination force test and an agglutination value test.The agglutination force test uses a positive sample under specified conditions, and measures the time (seconds) until the start of agglutination and the size of the aggregate. The time (seconds) it takes for the liquid to reach "lawn" is measured.In addition, for the agglutination titer test, the sample is serially diluted 2 times under specified conditions.
Calculate the agglutination titer from the highest dilution that shows positivity.
この際、アルブミン液抗体血清では凝集力試験と凝集価
試験に規定があるが、食塩法抗体血清では凝集価試験の
みである。従来のポリクローナル抗体を使用した抗り血
液型判定用血清は、単独でアルブミン液抗体血清および
食塩法抗体血清の両者の特性を満足することができない
が、本発明の抗り血液型判定用試薬では、この両者の特
性が満足され、どちらの方法でも使用することができる
。At this time, there are regulations for agglutination test and agglutination titer test for albumin liquid antibody serum, but only agglutination titer test is required for saline method antibody serum. Conventional anti-blood type determination serum using polyclonal antibodies cannot satisfy the characteristics of both albumin liquid antibody serum and saline method antibody serum alone, but the anti-blood type determination reagent of the present invention does not satisfy the characteristics of both albumin liquid antibody serum and saline antibody serum. , both of these characteristics are satisfied and either method can be used.
さらに、本発明の抗り血液型判定用試薬は、ＤＩ＋因子
の検出のための間接クームス試験を行うことができる。Furthermore, the reagent for determining blood type of the present invention can perform indirect Coombs test for detection of DI+ factor.
一般にＤｌ′因子は抗り抗体との反応が弱いものの総称
とされ、明瞭な凝集反応が認められなかった場合、Ｒｈ
ｏ （Ｄ）因子が陰性かどうかの確認が必要となる。こ
のような場合、本発明の抗り血液型判定用試薬では、間
接クームス試験を行い、その凝集の有無から真のＲｈｏ
（Ｄ）陰性について判定することができる。In general, Dl' factor is a general term for factors that react weakly with anti-antibodies, and if no clear agglutination reaction is observed, Rh
o It is necessary to confirm whether factor (D) is negative. In such cases, the indirect Coombs test is performed using the reagent for blood type determination of the present invention, and the true Rho is determined from the presence or absence of agglutination.
(D) A negative determination can be made.
本発明の抗り血液型判定用試薬は、モノクローナル抗体
を用いることにより、ポリクローナル抗体に比べて高い
特異性のものが永続的に産生できる。従って、本発明の
抗り血液型判定用試薬は優れた均質な特性のものとして
ロフト開運なく製造することができるという効果を有す
る。By using a monoclonal antibody, the reagent for blood type determination of the present invention can be permanently produced with higher specificity than polyclonal antibodies. Therefore, the reagent for blood type determination of the present invention has excellent homogeneous properties and has the advantage that it can be manufactured without any loft problems.
特に、抗Ｄモノクローナル抗体としてイムノグロブリン
サブクラスがＩｇＧタイプの抗Ｄモノクローナル抗体と
イムノグロブリンサブクラスがＩｇＭタイプの抗Ｄモノ
クローナル抗体とを併用することによって、Ｒｈｏ （
Ｄ）因子に対する凝集反応が増強されて実用上刃価が上
がり、かつアルブミン液抗体血清および食塩法抗体血清
の両者の特性を満足することができ、とちらの方法も使
用することができるという効果を有する。In particular, by using together an anti-D monoclonal antibody whose immunoglobulin subclass is IgG type and an anti-D monoclonal antibody whose immunoglobulin subclass is IgM type, Rho (
D) The effect of enhancing the agglutination reaction against the factor, increasing the practical blade value, and satisfying the characteristics of both the albumin liquid antibody serum and the saline method antibody serum, so that both methods can also be used. has.
また、本発明の抗り血液型判定用試薬は、Ｄυ因子の検
出のための間接クームス試験を行うことができるという
効果をも有する。これはＩｇＭタイプのモノクローナル
抗体では行うことができない。Furthermore, the reagent for determining blood type of the present invention also has the effect that indirect Coombs test for detection of Dυ factor can be performed. This cannot be done with monoclonal antibodies of the IgM type.
以上のことから明らかなように、本発明の抗り血液型判
定用試薬は臨床検査の分野に有用なものとして供するこ
とができる。As is clear from the above, the reagent for determining blood type of the present invention can be useful in the field of clinical testing.
以下に本発明の実施例を示すが、本発明はこれら実施例
に限定されるものではない。Examples of the present invention are shown below, but the present invention is not limited to these Examples.
実施例１
本発明の抗り血液型判定用試薬は次のようにして作製す
ることができる。まず抗Ｄモノクローナル抗体のイムノ
グロブリンサブクラスがＩｇＧタイプのバルクをとり、
それと同じ力価の抗Ｄモノクローナル抗体のイムノグロ
ブリンサブクラスがＩｇＭタイプのバルクを加える。こ
れにウシアルブミンを２０％の濃度になるように加え、
さらに防腐剤としてアジ化ナトリウムを０．１％の濃度
になるように加えたあと、除菌し無菌的にバイアルに小
分けして作製する。Example 1 The reagent for determining blood type of the present invention can be produced as follows. First, the immunoglobulin subclass of the anti-D monoclonal antibody takes on the bulk of the IgG type,
The same titer of anti-D monoclonal antibody immunoglobulin subclass adds bulk to the IgM type. Add bovine albumin to this to a concentration of 20%,
Furthermore, sodium azide is added as a preservative to a concentration of 0.1%, and then sterilized and aseptically divided into vials for preparation.
試験例１
本発明の抗り血液型判定用試薬の凝集力試験については
次のように実施した。まずＤ陽性血球の４０ｖ／ｖ％浮
遊液をウシアルブミン溶液を用いて作製し、載せガラス
上でこの２滴に実施例１で作製した抗り血液型判定用試
薬の１滴を混合し、凝集が開始するまでの時間を測定し
た（測定１）。Test Example 1 A coagulation force test of the reagent for blood type determination of the present invention was carried out as follows. First, a 40v/v% suspension of D-positive blood cells was prepared using a bovine albumin solution, and one drop of the anti-blood type determination reagent prepared in Example 1 was mixed with two drops of this on a mounting glass, and the aggregation was carried out. The time required for this to start was measured (Measurement 1).
更にその凝集塊の大きさが１鴎８に達した時間も測定し
た（測定２）、このとき対照として抗り血液型判定用試
薬の代わりに、抗Ｄモノクローナル抗体のイムノグロブ
リンサブクラスがＩｇＧタイプのものおよび抗Ｄポリク
ローナル抗体からなる市販のＤａｄｓ社製造品も同様に
試験した。Furthermore, we measured the time it took for the size of the aggregate to reach 18 mm (Measurement 2).At this time, as a control, instead of using a reagent for determining blood type, we used a reagent in which the immunoglobulin subclass of the anti-D monoclonal antibody was IgG type. A commercial Dads product consisting of anti-D polyclonal antibody and anti-D polyclonal antibody was similarly tested.
その結果は表１のようになり、本発明の抗り血液型判定
用試薬は凝集力試験において満足できる成績を示した。The results are shown in Table 1, and the reagent for determining blood type of the present invention showed satisfactory results in the agglutination test.
試験例２
本発明の抗り血液型判定用試薬のアルブミン液抗体血清
としての凝集価試験については次のように実施した。ま
ずＤ陽性血球の２ ｖ ／ ｖ％浮遊液をウシアルブミ
ン溶液を用いて作製する０次に実施例１で作製した抗り
血液型判定用試薬をＡＢ型血清で２０連続希釈し、それ
ぞれＯ，ｌ Ｉｄを試験管にとり、これに上記の血球浮
遊液０．１ ｍを加え混和する。これを遠心沈澱し凝集
を観察する。この凝集を示す最高希釈倍数から凝集価を
求める。このとき対照として抗Ｄモノクローナル抗体の
イムノグロブリンサブクラスがＩｇＧタイプのものおよ
び抗Ｄポリクローナル抗体からなる市販のＤａｄｅ社製
造品も同様に試験した。Test Example 2 An agglutination titer test of the anti-blood type determination reagent of the present invention as an albumin liquid antibody serum was carried out as follows. First, a 2 v/v% suspension of D-positive blood cells was prepared using a bovine albumin solution.Next, the reagent for determining the anti-blood type prepared in Example 1 was serially diluted 20 times with AB type serum, and diluted with O, respectively. l Take the Id in a test tube, add 0.1 ml of the above blood cell suspension, and mix. This is centrifuged for sedimentation and aggregation is observed. The agglutination value is determined from the highest dilution ratio that shows this agglutination. At this time, as controls, anti-D monoclonal antibodies whose immunoglobulin subclass was IgG type and commercially available anti-D polyclonal antibodies manufactured by Dade were also tested in the same manner.
その結果は表２のようになり、本発明の抗り血液型判定
用試薬はアルブミン液抗体血清としての凝集価試験にお
いても満足できる成績を示した。The results are shown in Table 2, and the anti-blood type determination reagent of the present invention showed satisfactory results in the agglutination titer test as an albumin liquid antibody serum.
（以下余白）
表２
試験例３
本発明の抗り血液型判定用試薬の食塩液抗体血清として
の凝集価試験については次のように実施した。まずＤ陽
性血球の２ｖ／ｖ％浮遊液を生理食塩液浮遊液を用いて
作製する０次に実施例１で作製した抗り血液型判定用試
薬を生理食塩液で２倍連続希釈し、それぞれ０．１ ｍ
を試験管にとり、これに上記の血球浮遊液０．１ ｍを
加え混和する。(The following is a blank space) Table 2 Test Example 3 The agglutination titer test of the anti-blood type determination reagent of the present invention as a saline antibody serum was carried out as follows. First, a 2v/v% suspension of D-positive blood cells was prepared using a physiological saline suspension.Next, the reagent for determining blood type determined in Example 1 was serially diluted 2 times with physiological saline, and each 0.1 m
Place in a test tube, add 0.1 ml of the above blood cell suspension, and mix.
これを遠心沈澱し凝集を観察する。この凝集を示す最高
希釈倍数から凝集価を求める。このとき対照として抗Ｄ
モノクローナル抗体のイムノグロブリンサブクラスがＩ
ｇＧタイプのものも同様に試験した。This is centrifuged for sedimentation and aggregation is observed. The agglutination value is determined from the highest dilution ratio that shows this agglutination. At this time, as a control, anti-D
The immunoglobulin subclass of monoclonal antibodies is I.
The gG type was also tested in the same way.
その結果は表３のようになり、本発明の抗り血液型判定
用試薬は食塩液抗体血清としての凝集価試験においても
満足できる成績を示した。The results are shown in Table 3, and the reagent for determining blood type of the present invention showed satisfactory results in the agglutination titer test as a saline antibody serum.
なお、この際、対象として用いた抗Ｄモノクローナル抗
体（ｒｇｃタイプ）は厚生省の製造基準に適合すること
が出来なかった。In addition, at this time, the anti-D monoclonal antibody (rgc type) used as a subject could not meet the manufacturing standards of the Ministry of Health and Welfare.
表３Table 3
Claims (1)
クローナル抗体、イムノグロブリンサブクラスがＩｇＭ
タイプの抗Ｄモノクローナル抗体および蛋白質よりなる
ことを特徴とするＲｈｏ（Ｄ）因子検出のための抗Ｄ血
液型判定用試薬。Anti-D monoclonal antibody whose immunoglobulin subclass is IgG type, immunoglobulin subclass is IgM
1. A reagent for anti-D blood type determination for detecting Rho (D) factor, characterized by comprising a type of anti-D monoclonal antibody and protein.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP63050710A JPH0623760B2 (en) | 1988-03-03 | 1988-03-03 | Anti-D blood typing reagent |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP63050710A JPH0623760B2 (en) | 1988-03-03 | 1988-03-03 | Anti-D blood typing reagent |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| JPH01223349A true JPH01223349A (en) | 1989-09-06 |
| JPH0623760B2 JPH0623760B2 (en) | 1994-03-30 |
Family
ID=12866451
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP63050710A Expired - Lifetime JPH0623760B2 (en) | 1988-03-03 | 1988-03-03 | Anti-D blood typing reagent |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPH0623760B2 (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113341163A (en) * | 2021-06-03 | 2021-09-03 | 上海市血液中心 | D-rare blood type screening reagent and application thereof |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS59132888A (en) * | 1982-12-30 | 1984-07-31 | ビオテスト アウチエンゲゼルシャフト | Monoclonal antibody reacted by direct aggultination test andpeculiar to humna blood antigen d and hybridoma cell line for producing same |
| JPS60136599A (en) * | 1983-12-26 | 1985-07-20 | Isao Ono | Monoclonal antibody |
| JPS6344881A (en) * | 1986-04-25 | 1988-02-25 | セントラル、ブラツド、ラボラトリ−ズ、オ−ソリテイ− | Heterohybridoma producing human anti-rhesus-d |
| JPS63127162A (en) * | 1986-06-16 | 1988-05-31 | Nippon Sekijiyuujishiya | Antigen-antibody composite and its use |
-
1988
- 1988-03-03 JP JP63050710A patent/JPH0623760B2/en not_active Expired - Lifetime
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS59132888A (en) * | 1982-12-30 | 1984-07-31 | ビオテスト アウチエンゲゼルシャフト | Monoclonal antibody reacted by direct aggultination test andpeculiar to humna blood antigen d and hybridoma cell line for producing same |
| JPS60136599A (en) * | 1983-12-26 | 1985-07-20 | Isao Ono | Monoclonal antibody |
| JPS6344881A (en) * | 1986-04-25 | 1988-02-25 | セントラル、ブラツド、ラボラトリ−ズ、オ−ソリテイ− | Heterohybridoma producing human anti-rhesus-d |
| JPS63127162A (en) * | 1986-06-16 | 1988-05-31 | Nippon Sekijiyuujishiya | Antigen-antibody composite and its use |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113341163A (en) * | 2021-06-03 | 2021-09-03 | 上海市血液中心 | D-rare blood type screening reagent and application thereof |
| CN113341163B (en) * | 2021-06-03 | 2024-01-19 | 上海市血液中心 | -D-rare blood group screening reagent and application thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| JPH0623760B2 (en) | 1994-03-30 |
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