CN1136230C - 泌尿生殖癌tlp复合体肽及其抗体 - Google Patents
泌尿生殖癌tlp复合体肽及其抗体 Download PDFInfo
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- CN1136230C CN1136230C CNB971912130A CN97191213A CN1136230C CN 1136230 C CN1136230 C CN 1136230C CN B971912130 A CNB971912130 A CN B971912130A CN 97191213 A CN97191213 A CN 97191213A CN 1136230 C CN1136230 C CN 1136230C
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- 108090000765 processed proteins & peptides Proteins 0.000 title claims abstract description 20
- 201000009030 Carcinoma Diseases 0.000 title abstract description 8
- 102000004196 processed proteins & peptides Human genes 0.000 title description 5
- 239000003153 chemical reaction reagent Substances 0.000 abstract description 5
- 125000003275 alpha amino acid group Chemical group 0.000 abstract description 4
- 239000008194 pharmaceutical composition Substances 0.000 abstract description 2
- 206010028980 Neoplasm Diseases 0.000 description 8
- 239000007983 Tris buffer Substances 0.000 description 4
- 201000011510 cancer Diseases 0.000 description 4
- 239000006228 supernatant Substances 0.000 description 4
- 210000001550 testis Anatomy 0.000 description 4
- LENZDBCJOHFCAS-UHFFFAOYSA-N tris Chemical compound OCC(N)(CO)CO LENZDBCJOHFCAS-UHFFFAOYSA-N 0.000 description 4
- 238000000034 method Methods 0.000 description 3
- 235000018102 proteins Nutrition 0.000 description 3
- 102000004169 proteins and genes Human genes 0.000 description 3
- 108090000623 proteins and genes Proteins 0.000 description 3
- 102000007474 Multiprotein Complexes Human genes 0.000 description 2
- 108010085220 Multiprotein Complexes Proteins 0.000 description 2
- 208000008385 Urogenital Neoplasms Diseases 0.000 description 2
- 235000001014 amino acid Nutrition 0.000 description 2
- 150000001413 amino acids Chemical class 0.000 description 2
- 201000006662 cervical adenocarcinoma Diseases 0.000 description 2
- 238000001962 electrophoresis Methods 0.000 description 2
- 238000001914 filtration Methods 0.000 description 2
- 238000002360 preparation method Methods 0.000 description 2
- 208000037964 urogenital cancer Diseases 0.000 description 2
- QKNYBSVHEMOAJP-UHFFFAOYSA-N 2-amino-2-(hydroxymethyl)propane-1,3-diol;hydron;chloride Chemical compound Cl.OCC(N)(CO)CO QKNYBSVHEMOAJP-UHFFFAOYSA-N 0.000 description 1
- 238000009007 Diagnostic Kit Methods 0.000 description 1
- 206010058467 Lung neoplasm malignant Diseases 0.000 description 1
- 239000002033 PVDF binder Substances 0.000 description 1
- 208000009956 adenocarcinoma Diseases 0.000 description 1
- 238000003277 amino acid sequence analysis Methods 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 230000000890 antigenic effect Effects 0.000 description 1
- 208000019065 cervical carcinoma Diseases 0.000 description 1
- 210000003679 cervix uteri Anatomy 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 238000003745 diagnosis Methods 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 230000005611 electricity Effects 0.000 description 1
- 238000010828 elution Methods 0.000 description 1
- 230000008014 freezing Effects 0.000 description 1
- 238000007710 freezing Methods 0.000 description 1
- 210000003734 kidney Anatomy 0.000 description 1
- 201000005202 lung cancer Diseases 0.000 description 1
- 208000020816 lung neoplasm Diseases 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 230000001338 necrotic effect Effects 0.000 description 1
- 229920001184 polypeptide Polymers 0.000 description 1
- 229920002981 polyvinylidene fluoride Polymers 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 239000000243 solution Substances 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
- 210000004881 tumor cell Anatomy 0.000 description 1
- 208000012991 uterine carcinoma Diseases 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
Classifications
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/46—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates
- C07K14/47—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals
- C07K14/4701—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals not used
- C07K14/4748—Tumour specific antigens; Tumour rejection antigen precursors [TRAP], e.g. MAGE
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
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- Molecular Biology (AREA)
- Biochemistry (AREA)
- Biophysics (AREA)
- Gastroenterology & Hepatology (AREA)
- Genetics & Genomics (AREA)
- Zoology (AREA)
- Toxicology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Immunology (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- General Chemical & Material Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Pharmacology & Pharmacy (AREA)
- Animal Behavior & Ethology (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Peptides Or Proteins (AREA)
- Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Investigating Or Analysing Biological Materials (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
Abstract
本文描述了含有特异泌尿生殖癌TLP表位的肽，其中TLP包含氨基酸序列：Gly Pro Pro Glu Val Gln Asn Ala Asn。另外也描述了检测所述泌尿生殖TLPs的试剂及药物组合物。
Description
本发明涉及从泌尿生殖器癌中分离出来的TLP复合体(肿瘤释放蛋白)的肽。
具体地讲，本发明涉及来自人泌尿生殖器的具有抗原活性的TLP蛋白复合体的肽，以及能与所述肽反应的抗体，所述抗体可用于诊断及临床。
TLP复合体是存在于人肿瘤细胞中的蛋白复合体。有人描述了一种214KDa的TLP蛋白(Tarro G.，Oncology 40，248-253，1983)。可用欧洲专利283443中所描述的方法来从肿瘤组织中分离TLP。意大利专利申请RM92A000506从肺癌中识别了一种TLP蛋白。本发明的发明人惊奇地发现来自泌尿生殖器癌的TLP含有新肽，这些新肽含有不同于已知TLP肽的序列。
因此，需要从泌尿生殖癌中鉴别出起表位作用的TLP肽，以制备作为抗体的特异试剂。
本发明的发明人已识别出具有一定序列的肽，所述序列包含于来自子宫颈或睾丸腺癌、以及肾瘤的214KDa的TLP蛋白的序列中。
因此本发明的一个目是提供含有特异性泌尿生殖癌TLP表位的肽，其中所述的TLP的特征是含有SEQ ID No.1的氨基酸序列：
Gly Pro Pro Glu Val Gln Asn Ala Asn
按照优选实施方案，其中的肽含有SEQ ID No.1的氨基酸序列。
本发明的再一目的是提供能从泌尿生殖癌中识别TLP的特异性试剂，所述试剂优选含有TLP抗体。更优选地，所述抗体可识别具有SEQID No.1序列的肽片断。
本发明的又一目的是提供诊断试剂盒，所述试剂盒含有本发明抗体作为特异性试剂，以从样品中识别TLP。
本发明的另一目的是提供含有本发明肽作为活性剂的药物组合物。
现在用说明性并非限制性的实例对本发明进行描述。实施例1 肿瘤浸出液的制备
将来自一患有子宫癌的45岁妇女的12.55g的肿瘤在室温下解冻，并且用外科方法除去坏死组织。当将所得到的物质匀化后，用Tris 1×(10mM Tris-HCl，pH7.2)进行多次洗涤，并且将该组织进行三轮冷冻-解冻循环。
收集使用过的Tris洗涤液并在33,000rpm下离心1小时。收集上清液并进行冷冻。
将组织用声波处理三次达3分钟，随后在33,000rpm下超离心120分钟。收集上清液(5.7ml)，在Agrodisc过滤器(0.45μm)上过滤，并将过滤后得的组织悬浮在Tris溶液中，比例为1g/ml Tris，并将其在33,000rpm下超离心60分钟。过滤上清液(0.5ml)并将其加入上一次的上清液中。
对睾丸癌(总产量1.9ml)及肾瘤样品进行同样的处理。实施例2 对来自泌尿生殖器癌的TLP蛋白中包含的肽的识别
如欧洲专利283443中所述，将TLP复合物从泌尿生殖器癌浸出液中分离出来。所使用的样品是：
1)子宫颈腺癌；
2)睾丸癌；
3)肾瘤。
所得到的蛋白含量如下：
表1
样品 总mg数 mg/ml TLP
子宫颈腺癌 59.5 9.6
睾丸癌 7.4 3.9
肾瘤 7.4* 1.5
*总肾蛋白浓度不允许足够的TLP纯化产量。
用变性电泳分析方法(SDS)，通过分子量测定(214KDa)来进行TLP识别。
通过取下相应于表观分子量214kDa的凝胶部分并进一步用电洗脱仪(AMICON)进行电洗脱以去除大部分的污染物。
随后通过变性电泳分析确定TLP的纯度。然后将TLP移至具有很高的蛋白质结合能力的PVDF(聚二氟乙烯(Polyvinylidifluoride))膜上，再用Applied Biosystems的蛋白质自动测序仪按照Edman方法进行氨基酸序列分析。
在所有样品中发现的氨基酸序列如下：
Gly Pro Pro Glu Val Gln Asn Ala Asn(SEQ ID No.1)
序列表(1)一般信息(i)申请人
(A)名称：Istituto Farmacoterapico Italiano S.p.A.
(B)街道：Via Paolo Frisi 21/23
(C)城市：罗马
(E)国家：意大利
(F)邮政编码(ZIP)：00197
(A)名称：Tarro Giulio c/o Istituto Farmacoterapica Italiano
S.p.A.
(B)街道：Via Paolo Frisi 21/23
(C)城市：罗马
(E)国家：意大利
(F)邮政编码(ZIP)：00197(ii)发明名称：泌尿生殖癌TLP复合体肽及其抗体(iii)序列数：1(iv)计算机可读形式：
(A)媒体类型：软盘
(B)计算机：IBM PC兼容机
(C)操作系统：PC-DOS/MS-DOS
(D)软件：Patentin Release#1.0，Version#1.30(EPO)(2)SEQ ID NO：1的信息：(i)序列特征：
(A)长度：9个氨基酸
(B)类型：氨基酸
(C)链型：单链
(D)拓朴学：线性(ii)分子类型：肽(xi)序列描述：SEQ ID NO：1：
Gly Pro Pro Glu Val Gln Asn Ala Asn
1 5
Claims (1)
1.一种泌尿生殖器TLP肽，所述肽具有SEQ ID No.1序列。
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| ITRM96A000496 | 1996-07-10 | ||
| IT96RM000496A IT1284870B1 (it) | 1996-07-10 | 1996-07-10 | Peptidi dei complessi tlp da carcinomi dell'apparato urogenitale e anticorpi diretti contro di essi |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN1205012A CN1205012A (zh) | 1999-01-13 |
| CN1136230C true CN1136230C (zh) | 2004-01-28 |
Family
ID=11404336
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CNB971912130A Expired - Fee Related CN1136230C (zh) | 1996-07-10 | 1997-07-02 | 泌尿生殖癌tlp复合体肽及其抗体 |
Country Status (16)
| Country | Link |
|---|---|
| US (3) | US6222010B1 (zh) |
| EP (1) | EP0851872B1 (zh) |
| CN (1) | CN1136230C (zh) |
| AT (1) | ATE200678T1 (zh) |
| AU (1) | AU716721B2 (zh) |
| CA (1) | CA2234151C (zh) |
| DE (1) | DE69704606T2 (zh) |
| DK (1) | DK0851872T3 (zh) |
| ES (1) | ES2156393T3 (zh) |
| GR (1) | GR3036057T3 (zh) |
| IT (1) | IT1284870B1 (zh) |
| NO (1) | NO982227D0 (zh) |
| PT (1) | PT851872E (zh) |
| RU (1) | RU2203679C2 (zh) |
| SK (1) | SK47098A3 (zh) |
| WO (1) | WO1998001462A2 (zh) |
Families Citing this family (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| IT1284870B1 (it) | 1996-07-10 | 1998-05-22 | Ist Farmacoterapico It Spa | Peptidi dei complessi tlp da carcinomi dell'apparato urogenitale e anticorpi diretti contro di essi |
| DE10001717C1 (de) * | 2000-01-18 | 2001-04-26 | Xcellsis Gmbh | Brennstoffzellensystem |
| ITMI20011380A1 (it) * | 2001-06-29 | 2002-12-29 | Unihart Corp | Antigene tlp e sue applicazioni diagnostiche |
| CN101712946B (zh) * | 2009-09-27 | 2013-02-27 | 南通大学附属医院 | 构建生殖结节器官培养尿道体外发育模型的方法 |
Family Cites Families (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| IT1206250B (it) | 1987-02-18 | 1989-04-14 | Ist Farmacoterapico It Spa | Stiche umane e relativi prodotti metodo per l'estrazione di antigeni di membrana aventi proprieta' immunogene a partire da cellule neopla- |
| IT1262954B (it) | 1992-07-03 | 1996-07-23 | Ist Farmacoterapico It Spa | Regioni antigeniche dei complessi tpl e anticorpi diretti contro di essi. |
| IT1284870B1 (it) | 1996-07-10 | 1998-05-22 | Ist Farmacoterapico It Spa | Peptidi dei complessi tlp da carcinomi dell'apparato urogenitale e anticorpi diretti contro di essi |
-
1996
- 1996-07-10 IT IT96RM000496A patent/IT1284870B1/it active IP Right Grant
-
1997
- 1997-07-02 WO PCT/IT1997/000158 patent/WO1998001462A2/en active IP Right Grant
- 1997-07-02 PT PT97932002T patent/PT851872E/pt unknown
- 1997-07-02 AU AU35572/97A patent/AU716721B2/en not_active Ceased
- 1997-07-02 CN CNB971912130A patent/CN1136230C/zh not_active Expired - Fee Related
- 1997-07-02 RU RU98106385/14A patent/RU2203679C2/ru not_active IP Right Cessation
- 1997-07-02 CA CA002234151A patent/CA2234151C/en not_active Expired - Fee Related
- 1997-07-02 US US09/051,357 patent/US6222010B1/en not_active Expired - Fee Related
- 1997-07-02 AT AT97932002T patent/ATE200678T1/de not_active IP Right Cessation
- 1997-07-02 SK SK470-98A patent/SK47098A3/sk unknown
- 1997-07-02 DE DE69704606T patent/DE69704606T2/de not_active Expired - Fee Related
- 1997-07-02 DK DK97932002T patent/DK0851872T3/da active
- 1997-07-02 EP EP97932002A patent/EP0851872B1/en not_active Expired - Lifetime
- 1997-07-02 ES ES97932002T patent/ES2156393T3/es not_active Expired - Lifetime
-
1998
- 1998-05-15 NO NO982227A patent/NO982227D0/no unknown
-
2000
- 2000-02-15 US US09/504,089 patent/US20030170750A1/en not_active Abandoned
- 2000-09-14 US US09/661,503 patent/US6664378B1/en not_active Expired - Fee Related
-
2001
- 2001-06-14 GR GR20010400899T patent/GR3036057T3/el not_active IP Right Cessation
Also Published As
| Publication number | Publication date |
|---|---|
| PT851872E (pt) | 2001-08-30 |
| CA2234151C (en) | 2004-02-03 |
| CA2234151A1 (en) | 1998-01-15 |
| ITRM960496A1 (it) | 1998-01-10 |
| US20030170750A1 (en) | 2003-09-11 |
| EP0851872A3 (en) | 1998-10-14 |
| WO1998001462A2 (en) | 1998-01-15 |
| NO982227L (no) | 1998-05-15 |
| RU2203679C2 (ru) | 2003-05-10 |
| WO1998001462A3 (en) | 1998-08-06 |
| US6222010B1 (en) | 2001-04-24 |
| DE69704606D1 (de) | 2001-05-23 |
| ES2156393T3 (es) | 2001-06-16 |
| DE69704606T2 (de) | 2001-11-08 |
| DK0851872T3 (da) | 2001-06-18 |
| IT1284870B1 (it) | 1998-05-22 |
| SK47098A3 (en) | 1999-02-11 |
| EP0851872A2 (en) | 1998-07-08 |
| CN1205012A (zh) | 1999-01-13 |
| GR3036057T3 (en) | 2001-09-28 |
| NO982227D0 (no) | 1998-05-15 |
| AU3557297A (en) | 1998-02-02 |
| AU716721B2 (en) | 2000-03-02 |
| ATE200678T1 (de) | 2001-05-15 |
| EP0851872B1 (en) | 2001-04-18 |
| US6664378B1 (en) | 2003-12-16 |
| ITRM960496A0 (zh) | 1996-07-10 |
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| Date | Code | Title | Description |
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| C06 | Publication | ||
| PB01 | Publication | ||
| C53 | Correction of patent of invention or patent application | ||
| COR | Change of bibliographic data |
Free format text: CORRECT: APPLICANT; FROM: ISTITUTO FARMACEOTERAPICO ITALIANO S. P. A. TO: UNIHART CORP |
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| CP03 | Change of name, title or address |
Address after: Dublin, Ireland Applicant after: Unelhatt Corp. Address before: Rome Italy Applicant before: Istituto Farmaceoterapico Italiano S. P. A. |
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