JPH0570438B2 - - Google Patents
Info
- Publication number
- JPH0570438B2 JPH0570438B2 JP61189810A JP18981086A JPH0570438B2 JP H0570438 B2 JPH0570438 B2 JP H0570438B2 JP 61189810 A JP61189810 A JP 61189810A JP 18981086 A JP18981086 A JP 18981086A JP H0570438 B2 JPH0570438 B2 JP H0570438B2
- Authority
- JP
- Japan
- Prior art keywords
- ala
- arg
- peptide
- phe
- monoclonal antibody
- 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 - Fee Related
Links
- 108090000765 processed proteins & peptides Proteins 0.000 claims description 16
- 102000054727 Serum Amyloid A Human genes 0.000 claims description 6
- 108700028909 Serum Amyloid A Proteins 0.000 claims description 6
- 210000004408 hybridoma Anatomy 0.000 claims description 4
- 210000004027 cell Anatomy 0.000 claims description 3
- PHJPKNUWWHRAOC-PEFMBERDSA-N Asn-Ile-Gln Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CCC(=O)N)C(=O)O)NC(=O)[C@H](CC(=O)N)N PHJPKNUWWHRAOC-PEFMBERDSA-N 0.000 claims description 2
- 102000014914 Carrier Proteins Human genes 0.000 claims description 2
- 108010078791 Carrier Proteins Proteins 0.000 claims description 2
- 206010003445 Ascites Diseases 0.000 claims 3
- 241000124008 Mammalia Species 0.000 claims 3
- JZDHUJAFXGNDSB-WHFBIAKZSA-N Glu-Ala Chemical compound OC(=O)[C@H](C)NC(=O)[C@@H](N)CCC(O)=O JZDHUJAFXGNDSB-WHFBIAKZSA-N 0.000 claims 1
- 241001465754 Metazoa Species 0.000 claims 1
- 206010035226 Plasma cell myeloma Diseases 0.000 claims 1
- 108010049041 glutamylalanine Proteins 0.000 claims 1
- 230000003053 immunization Effects 0.000 claims 1
- 201000000050 myeloid neoplasm Diseases 0.000 claims 1
- 210000003200 peritoneal cavity Anatomy 0.000 claims 1
- 210000004989 spleen cell Anatomy 0.000 claims 1
- 206010002022 amyloidosis Diseases 0.000 description 9
- 150000001413 amino acids Chemical group 0.000 description 6
- 208000023769 AA amyloidosis Diseases 0.000 description 4
- 206010039811 Secondary amyloidosis Diseases 0.000 description 4
- 102000009091 Amyloidogenic Proteins Human genes 0.000 description 3
- 108010048112 Amyloidogenic Proteins Proteins 0.000 description 3
- 201000010099 disease Diseases 0.000 description 3
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 3
- 206010016202 Familial Amyloidosis Diseases 0.000 description 2
- 208000017105 hereditary amyloidosis Diseases 0.000 description 2
- 238000010827 pathological analysis Methods 0.000 description 2
- 102000004196 processed proteins & peptides Human genes 0.000 description 2
- VZQHRKZCAZCACO-PYJNHQTQSA-N (2s)-2-[[(2s)-2-[2-[[(2s)-2-[[(2s)-2-amino-5-(diaminomethylideneamino)pentanoyl]amino]propanoyl]amino]prop-2-enoylamino]-3-methylbutanoyl]amino]propanoic acid Chemical compound OC(=O)[C@H](C)NC(=O)[C@H](C(C)C)NC(=O)C(=C)NC(=O)[C@H](C)NC(=O)[C@@H](N)CCCNC(N)=N VZQHRKZCAZCACO-PYJNHQTQSA-N 0.000 description 1
- 208000023761 AL amyloidosis Diseases 0.000 description 1
- SVBXIUDNTRTKHE-CIUDSAMLSA-N Ala-Arg-Glu Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCC(O)=O)C(O)=O SVBXIUDNTRTKHE-CIUDSAMLSA-N 0.000 description 1
- MNBHKGYCLBUIBC-UFYCRDLUSA-N Arg-Phe-Phe Chemical compound C([C@H](NC(=O)[C@H](CCCNC(N)=N)N)C(=O)N[C@@H](CC=1C=CC=CC=1)C(O)=O)C1=CC=CC=C1 MNBHKGYCLBUIBC-UFYCRDLUSA-N 0.000 description 1
- OBMZMSLWNNWEJA-XNCRXQDQSA-N C1=CC=2C(C[C@@H]3NC(=O)[C@@H](NC(=O)[C@H](NC(=O)N(CC#CCN(CCCC[C@H](NC(=O)[C@@H](CC4=CC=CC=C4)NC3=O)C(=O)N)CC=C)NC(=O)[C@@H](N)C)CC3=CNC4=C3C=CC=C4)C)=CNC=2C=C1 Chemical compound C1=CC=2C(C[C@@H]3NC(=O)[C@@H](NC(=O)[C@H](NC(=O)N(CC#CCN(CCCC[C@H](NC(=O)[C@@H](CC4=CC=CC=C4)NC3=O)C(=O)N)CC=C)NC(=O)[C@@H](N)C)CC3=CNC4=C3C=CC=C4)C)=CNC=2C=C1 OBMZMSLWNNWEJA-XNCRXQDQSA-N 0.000 description 1
- 206010016207 Familial Mediterranean fever Diseases 0.000 description 1
- 208000005531 Immunoglobulin Light-chain Amyloidosis Diseases 0.000 description 1
- 241000699670 Mus sp. Species 0.000 description 1
- 101710176384 Peptide 1 Proteins 0.000 description 1
- 206010036673 Primary amyloidosis Diseases 0.000 description 1
- 230000000890 antigenic effect Effects 0.000 description 1
- 230000008021 deposition Effects 0.000 description 1
- 239000000835 fiber Substances 0.000 description 1
- 230000009760 functional impairment Effects 0.000 description 1
- 230000002163 immunogen Effects 0.000 description 1
- 238000011835 investigation Methods 0.000 description 1
- 210000000056 organ Anatomy 0.000 description 1
- 208000008798 osteoma Diseases 0.000 description 1
- 230000001575 pathological effect Effects 0.000 description 1
- 108010084525 phenylalanyl-phenylalanyl-glycine Proteins 0.000 description 1
- 230000000750 progressive effect Effects 0.000 description 1
- 230000009257 reactivity Effects 0.000 description 1
- 238000010561 standard procedure Methods 0.000 description 1
Description
産業上の利用分野
本発明は、ヒトのアミロイドＡ蛋白質に対する
単クローン性抗体に関する。本発明の単クローン
性抗体は続発性アミロイド−シスの検出に有用で
ある。
従来技術
アミロイドＡ蛋白質に対する単クローン性抗体
としては、ザ・ジヤーナル・オブ・ヒストケミス
トリー・エンド・サイトケミストー（J.
Histochem.Cytochem.）32，322〜328，1984に
報告があるが、アミロイド−シス患者組織から精
製したアミロイドＡ蛋白質を免疫源に用いてお
り、抗体の認織する抗原部位（エピトープ）は明
らかでない。
発明が解決しようとする問題
アミロイド−シスは、不溶性のアミロイド蛋白
質が種々の組織に沈着し、アミロイド線維やアミ
ロイド班を形成し、その沈着した組織（臓器）の
機能障害を引き起こし、進行性に病態が悪化し、
ついには死に到る難病である。昭和50年に厚生省
特定疾患アミロイド−シス調査研究班が組織さ
れ、病理学的に、(1)原発性アミロイド−シス、(2)
多発性骨随腫に合併するアミロイド−シス、(3)続
発性アミロイド−シス、(4)分類困難なアミロイド
−シス、(5)遺伝性アミロイド−シスおよび、(6)限
局性アミロイド−シスに分類されている。その分
類によつて、沈着するアミロイド蛋白質の種類も
異なり、アミロイドＡ蛋白質は、続発性アミロイ
ド−シスのほとんどすべてと、家族性地中海熱に
伴うアミロイド−シス（遺伝性アミロイド−シ
ス）の一部に沈着が認められる。一種類の単クロ
ーン性抗体によつて、全てのアミロイド−シスの
病理診断を行うことは、原理的にも不可能である
が、それぞれのアミロイド蛋白質に特異的な単ク
ローン性抗体を確立すば、アミロイド−シスの病
理診断上有用である。
本発明者は、続発性アミロイド−シスを検出す
るのに有用な単クローン性抗体を製造すべく研究
を行つた。すなわち、下記第１表で示されるヒト
のアミロイドＡ蛋白質の中から、１〜12番目のア
ミノ酸配列で示されるペプチド（以下ペプチド１
という）、15〜25番目のアミノ酸配列で示される
ペプチド（以下ペプチド２という）、37〜47番目
のアミノ酸配列で示されるペプチド（以下ペプチ
ド３という）、56〜66番目のアミノ酸配列で示さ
れるペプチド（以下ペプチド４という）および66
〜76番目のアミノ酸配列で示されるペプチド（以
下ペプチド５という）を合成し、これらをそれぞ
れ担体蛋白質と結合させ、結合物を用いてマウス
を免疫し、常法に従つて単クローン性抗体産生ハ
イブリドーマ細胞株を造成した。その結果、ペプ
チド３には反応するが、ペプチド１，２，４およ
び５には実質的に反応性を示さない単クローン性
抗体を生産するハイブリドーマ細胞株を得、本発
明を完成するに至つた。
INDUSTRIAL APPLICATION FIELD OF THE INVENTION The present invention relates to monoclonal antibodies against human amyloid A protein. The monoclonal antibodies of the invention are useful for detecting secondary amyloidosis. Prior Art Monoclonal antibodies against amyloid A protein are available from The Journal of Histochemistry and Cytochemistry (J.
Histochem.Cytochem.) 32 , 322-328, 1984, but amyloid A protein purified from amyloidosis patient tissue is used as an immunogen, and the antigenic site (epitope) to which the antibody is identified is unclear. . Problems to be Solved by the Invention Amyloidosis is a disease in which insoluble amyloid protein is deposited in various tissues, forming amyloid fibers and plaques, causing functional impairment of the deposited tissues (organs), and leading to a progressive pathological condition. worsens,
It is an incurable disease that eventually leads to death. In 1975, the Ministry of Health and Welfare's specified disease amyloidosis investigation research team was organized, and pathologically investigated (1) primary amyloidosis, (2)
Amyloidosis associated with multiple osteoma, (3) secondary amyloidosis, (4) amyloidosis that is difficult to classify, (5) hereditary amyloidosis, and (6) localized amyloidosis. Classified. Depending on the classification, the type of amyloid protein deposited differs, and amyloid A protein is present in almost all secondary amyloidosis and some amyloidosis associated with familial Mediterranean fever (hereditary amyloidosis). Deposition is observed. Although it is impossible in principle to perform pathological diagnosis of all amyloidosis using one type of monoclonal antibody, it is possible to establish monoclonal antibodies specific to each amyloid protein. , is useful in the pathological diagnosis of amyloidosis. The inventor conducted research to produce monoclonal antibodies useful for detecting secondary amyloidosis. That is, from among the human amyloid A proteins shown in Table 1 below, the peptides shown by the 1st to 12th amino acid sequences (hereinafter referred to as peptide 1)
), the peptide represented by the amino acid sequence 15th to 25th (hereinafter referred to as peptide 2), the peptide represented by the amino acid sequence 37th to 47th (hereinafter referred to as peptide 3), the peptide represented by the amino acid sequence 56th to 66th. (hereinafter referred to as peptide 4) and 66
Synthesize the peptide shown by the amino acid sequence of ~76th amino acid (hereinafter referred to as peptide 5), bind these to carrier proteins, immunize mice with the conjugate, and generate monoclonal antibody-producing hybridomas according to standard methods. A cell line was created. As a result, they obtained a hybridoma cell line that produces a monoclonal antibody that reacts with peptide 3 but has substantially no reactivity with peptides 1, 2, 4, and 5, leading to the completion of the present invention. .
【表】
61 62 63 64 65 66 67 68 69 70
Ala Arg Glu Asn Ile Gln Arg Phe Phe Gly
【table】
61 62 63 64 65 66 67 68 69 70
Ala Arg Glu Asn Ile Gln Arg Phe Phe Gly
Claims (1)
arg−Gly−Asn−Tyr−Asp−Ala−Ala−Lys−
Argで示されるペプチドと反応し、かつArg−
Ser−Phe−Phe−Ser−Phe−Leu−Gly−Glu−
Ala−Phe−Asp，Arg−Asp−Met−Trp−Arg
−Ala−Tyr−Ser−Asn−Met−Arg，Glu−Ala
−Ile−Ser−Asp−Ala−Arg−Glu−Asn−Ile−
GlnおよびGln−Arg−Phe−Phe−Gly−His−
Gly−Ala−Glu−Asn−Serで示されるペプチド
とは実質的に反応しない単クローン性抗体。 ２ IgGまたはIgMクラスに属する特許請求の範
囲第１項記載の単クローン性抗体。 ３ His−Ala−Arg−Gly−Asn−Tyr−Asp−
Ala−Ala−Lys−Argで示されるペプチドまたは
その担体蛋白質との結合物で哺乳動物を免疫し、
該免疫動物の脾細胞と哺乳動物の滑髄腫細胞とを
融合させ、得られるハイブリドーマ細胞株から該
ペプチドに反応するハイブリドーマ細胞株を選
び、該細胞株を培地に培養するか、哺乳動物の腹
腔に投与して腹水化し、培養物または腹水中に該
ペプチドに反応する単クローン性抗体を蓄積さ
せ、該培養物または腹水から該単クローン性抗体
を採取することによつて得られる特許請求の範囲
第１項記載の単クローン性抗体。[Claims] 1. Reacts with amyloid A protein, His-Ala-
arg−Gly−Asn−Tyr−Asp−Ala−Ala−Lys−
Reacts with the peptide represented by Arg, and Arg−
Ser−Phe−Phe−Ser−Phe−Leu−Gly−Glu−
Ala−Phe−Asp, Arg−Asp−Met−Trp−Arg
−Ala−Tyr−Ser−Asn−Met−Arg, Glu−Ala
−Ile−Ser−Asp−Ala−Arg−Glu−Asn−Ile−
Gln and Gln−Arg−Phe−Phe−Gly−His−
A monoclonal antibody that does not substantially react with the peptide represented by Gly-Ala-Glu-Asn-Ser. 2. The monoclonal antibody according to claim 1, which belongs to the IgG or IgM class. 3 His−Ala−Arg−Gly−Asn−Tyr−Asp−
immunizing a mammal with a peptide represented by Ala-Ala-Lys-Arg or its conjugate with a carrier protein,
The spleen cells of the immunized animal and the myeloma cells of the mammal are fused, and a hybridoma cell line that reacts with the peptide is selected from the resulting hybridoma cell lines, and the cell line is cultured in a medium or injected into the peritoneal cavity of the mammal. Claims obtained by administering the peptide to ascites, accumulating a monoclonal antibody that reacts with the peptide in a culture or ascites, and collecting the monoclonal antibody from the culture or ascites. Monoclonal antibody according to item 1.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP61189810A JPS6344895A (en) | 1986-08-13 | 1986-08-13 | Anti-amyloid a protein monoclonal antibody |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP61189810A JPS6344895A (en) | 1986-08-13 | 1986-08-13 | Anti-amyloid a protein monoclonal antibody |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| JPS6344895A JPS6344895A (en) | 1988-02-25 |
| JPH0570438B2 true JPH0570438B2 (en) | 1993-10-05 |
Family
ID=16247585
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP61189810A Granted JPS6344895A (en) | 1986-08-13 | 1986-08-13 | Anti-amyloid a protein monoclonal antibody |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPS6344895A (en) |
Families Citing this family (12)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| BE1003316A5 (en) * | 1990-11-27 | 1992-02-25 | Will L F & Cie Sa | MONOCLONAL ANTIBODY USEFUL FOR THE DIAGNOSIS OF ALZHEIMER'S DISEASE, SECRETARY HYBRIDOMA OF SUCH A MONOCLONAL ANTIBODY AND PROCESS FOR PREPARING THE SAME. |
| AU711163B2 (en) * | 1995-07-21 | 1999-10-07 | Provost, Fellows And Scholars Of The College Of The Holy And Undivided Trinity Of Queen Elizabeth Near Dublin, The | Method for the quantitative measurement of human acute phase serum amyloid a protein; recombinant protein; specific antibody |
| US20070021345A1 (en) | 2003-06-30 | 2007-01-25 | Ehud Gazit | Peptides antibodies directed thereagainst and methods using same for diagnosing and treating amyloid-associated diseases |
| US20040052928A1 (en) | 2002-09-06 | 2004-03-18 | Ehud Gazit | Peptides and methods using same for diagnosing and treating amyloid-associated diseases |
| JP2006506942A (en) * | 2002-01-31 | 2006-03-02 | テル アヴィヴ ユニヴァーシティ フューチャー テクノロジー ディヴェロップメント エル．ピー． | Peptides for diagnosing and treating amyloid-related diseases, antibodies thereto, and methods of use thereof |
| US7491699B2 (en) | 2002-12-09 | 2009-02-17 | Ramot At Tel Aviv University Ltd. | Peptide nanostructures and methods of generating and using the same |
| ATE426575T1 (en) | 2003-01-07 | 2009-04-15 | Univ Ramot | PEPTIDE ANOSTRUCTURES CONTAINING FOREIGN MATERIAL AND METHOD FOR PRODUCING THE SAME |
| WO2005027901A1 (en) | 2003-09-25 | 2005-03-31 | Tel Aviv University Future Technology Development L.P. | Compositions and methods using same for treating amyloid-associated diseases |
| US7732479B2 (en) | 2004-08-19 | 2010-06-08 | Tel Aviv University Future Technology Development L.P. | Compositions for treating amyloid associated diseases |
| US7786086B2 (en) | 2004-09-08 | 2010-08-31 | Ramot At Tel-Aviv University Ltd. | Peptide nanostructures containing end-capping modified peptides and methods of generating and using the same |
| SG186689A1 (en) * | 2007-12-28 | 2013-01-30 | Univ Tennessee Res Foundation | Treatment and prophylaxis of amyloidosis |
| CA2817830A1 (en) | 2010-11-15 | 2012-05-24 | Ramot At Tel Aviv University Ltd. | Dipeptide analogs for treating conditions associated with amyloid fibril formation |
-
1986
- 1986-08-13 JP JP61189810A patent/JPS6344895A/en active Granted
Also Published As
| Publication number | Publication date |
|---|---|
| JPS6344895A (en) | 1988-02-25 |
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Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| LAPS | Cancellation because of no payment of annual fees |