RU2224766C2 - Способ получения рецепторов для человеческих антигенов и их применение - Google Patents
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- 102000036639 antigens Human genes 0.000 title claims abstract 36
- 108091007433 antigens Proteins 0.000 title claims abstract 36
- 238000000034 method Methods 0.000 title claims abstract 21
- 210000003719 b-lymphocyte Anatomy 0.000 claims abstract 8
- 108060003951 Immunoglobulin Proteins 0.000 claims abstract 6
- 239000012634 fragment Substances 0.000 claims abstract 6
- 102000018358 immunoglobulin Human genes 0.000 claims abstract 6
- 206010028980 Neoplasm Diseases 0.000 claims abstract 5
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- 108010079585 Immunoglobulin Subunits Proteins 0.000 claims 1
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- C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
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- C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
- C07K16/28—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants
- C07K16/30—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants from tumour cells
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- C07K—PEPTIDES
- C07K2317/00—Immunoglobulins specific features
- C07K2317/30—Immunoglobulins specific features characterized by aspects of specificity or valency
- C07K2317/34—Identification of a linear epitope shorter than 20 amino acid residues or of a conformational epitope defined by amino acid residues
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Abstract
Изобретение относится к области иммунобиотехнологии и может найти применение в медицине. Изобретение касается способа получения рецептора для человеческого антигена, который является низкоиммуногенным или неиммуногенным у людей, включающий в себя стадии, на которых отбирают комбинацию функционально перегруппированных VH и VL цепей иммуноглобулина, где по меньшей мере указанная VH цепь получена из по существу непримированных зрелых человеческих В-лимфоцитов или из по существу анергических человеческих В клеток, а указанная VL цепь получена из природной популяции человеческих В клеток, причем указанные цепи экспрессируются с рекомбинантного вектора и для отбора используют in vitro дисплей-систему для связывания с человеческим антигеном. Описаны также рецепторы, которые являются низкоиммуногенными или неиммуногенными у людей и направлены на человеческие антигены, причем указанные рецепторы можно получить с помощью способа по изобретению. Рецепторы предпочтительно представляют собой антитела или их фрагменты, либо иммуноконъюгаты, содержащие VH/VL цепи указанного антитела. В частности, описываются рецепторы, направленные на человеческие опухолевые антигены, предпочтительно на человеческий опухолевый антиген 17-1 А, также известный как ЕрСАМ, EGP или GA 733-2. Создан набор, полезный для осуществления способа по изобретению, и фармацевтическая композиция, способная конъюгировать антигены в человеческом организме, содержащие вышеупомянутые рецепторы. 5 с. и 21 з.п. ф-лы, 55 ил., 3 табл.
Description
Текст описания в факсимильном виде (см. графическую часть)н
Claims (26)
1. Способ получения рецептора для человеческого антигена, который низкоиммуногенен или неиммуногенен у людей, включающий стадию отбора комбинации функционально перегруппированных VH и VL цепей иммуноглобулина, отличающийся тем, что по меньшей мере VH цепь получают из непримированных зрелых человеческих В-лимфоцитов, а VL цепь - из природного репертуара человеческих В клеток, причем указанные цепи экспрессируются с рекомбинантного вектора, используя для отбора дисплей-систему in vitro для связывания с человеческим антигеном.
2. Способ по п.1, отличающийся тем, что рецептор для человеческого антигена представляет собой иммуноглобулин или его фрагмент.
3. Способ по п.2, отличающийся тем, что фрагмент иммуноглобулина представляет собой Fv-фрагмент.
4. Способ по любому из пп.1-3, отличающийся тем, что человеческий антиген представляет собой человеческий опухолевый антиген.
5. Способ по любому из пп.1-4, отличающийся тем, что человеческий опухолевый антиген представляет собой человеческий 17-1А антиген.
6. Способ по любому из пп.1-5, отличающийся тем, что дисплей-система in vitro представляет собой фаговую дисплей-систему.
7. Способ по любому из пп.1-6, отличающийся тем, что VH и возможно VL цепи иммуноглобулина получают из репертуара человеческого IgD.
8. Способ по любому из пп.1-7, отличающийся тем, что указанная комбинация перегруппированных цепей экспрессируется с одной или более различных библиотек.
9. Способ по любому из пп.1-8, отличающийся тем, что VH цепь имеет одну из двух аминокислотных последовательностей, описанных в SEQ ID No 143 (аминокислотные положения со 108 по 234) и SEQ ID No 145 (аминокислотные положения со 139 по 251), и/или VI цепь имеет одну из двух аминокислотных последовательностей, описанных в SEQ ID No 141 (аминокислотные положения с 1 по 107) и SEQ ID No 147 (аминокислотные положения со 125 по 231).
10. Способ по любому из пп.1-9, отличающийся тем, что отбор включает (1) связывание дисплей-носителя, экспрессирующего рецептор антигена (а) на иммобилизованном антигене-мишени или его фрагментах, (б) на возможно меченых клетках, экспрессирующих антиген-мишень или его фрагменты, или (в) с растворимым, предпочтительно меченым антигеном-мишенью или его фрагментами, (2) отмывку неспецифически связывающегося дисплей-носителя (а и б) и последующую элюцию специфически связывающегося дисплей-носителя или (3) позитивное обогащение дисплей-носителя, связанного с антигеном-мишенью (б и в), из раствора антигена-мишени или из суспензий клеток, экспрессирующих антиген-мишень, выделенные дисплей-носители, включающие рецепторы антигена, при необходимости размножают репликацией и подвергают дальнейшему отбору in vitro соответствии со стадиями (1)-(3).
11. Способ по любому из пп.1-10, отличающийся тем, что перед стадией отбора VH или VL цепь отбирают по способности связывания с антигеном вместе с заместительной V цепью.
12. Способ по п.11, отличающийся тем, что заместительная V цепь представляет собой мышиную VH или VL цепь.
13. Способ по любому из пп.1-12, отличающийся тем, что отбор, подходящей комбинации VH и VL цепей осуществляют путем тестирования одной и той же VH или VL цепи в комбинации со множеством различных VL или VH цепей на способность связывания с человеческим антигеном.
14. Способ по любому из пп.1-13, отличающийся тем, что после отбора дополнительно получают человеческие VH или VL цепи или соответствующие кодирующие их нуклеиновые кислоты и сливают эти цепи с теми же или другими VH или VL цепями, либо с константными областями тяжелой (СН) или легкой (CL) цепей иммуноглобулина или с их участками, либо с неиммуноглобулиновыми цепями и соответствующими нуклеиновыми кислотами соответственно, и/или после отбора дополнительно получают человеческие VH и VL цепи и физически соединяют эти цепи с небелковыми фармацевтическими средствами и/или другими биологически активными молекулами.
15. Способ по п.14, отличающийся тем, что константные области СН и CL цепей получены из человеческого IgGI или IgGS.
16. Рецептор для человеческого антигена, который является низкоиммуногенным или неиммуногенным у людей, содержащий комбинацию функционально перегруппированных VH и VL цепей, где по меньшей мере указанная VH цепь получена из непримированных зрелых человеческих В-лимфоцитов, а указанная VL цепь получена из природного репертуара человеческих B клеток.
17. Рецептор для человеческого антигена по п.16, отличающийся тем, что он получен способом по любому из пп.1-15.
18. Рецептор для человеческого антигена по п.16 или 17, отличающийся тем, что этот рецептор представляет собой антитело или его фрагмент.
19. Рецептор для человеческого антигена по любому из пп.16-18, отличающийся тем, что этот рецептор является специфичным для человеческого опухолевого антигена.
20. Рецептор для человеческого антигена по любому из пп.16-19, отличающийся тем, что этот рецептор является специфичным для нативного человеческого 17-1А антигена.
21. Рецептор для человеческого антигена по п.20, отличающийся тем, что VH цепь имеет одну из двух аминокислотных последовательностей, описанных SEQ ID No 143 (аминокислотные положения со 108 по 234) и SEQ ID No 145 (аминокислотные положения со 139 по 251), и/или VL цепь имеет одну из двух аминокислотных последовательностей, описанных SEQ ID No 141 (аминокислотные положения с 1 по 107) и SEQ ID No 147 (аминокислотные положения со 125 по 231).
22. Набор для отбора рецептора для человеческого антигена, содержащий комбинацию функционально перегруппированных VH и VL цепей иммуноглобулина, отличающийся тем, что по меньшей мере одна из VH и VL цепей получена из непримированных зрелых человеческих В-лимфоцитов, причем указанные цепи экспрессируются с рекомбинантных векторов in vitro дисплей-системы.
23. Рецептор для человеческого антигена, распознающий эпитоп внеклеточного домена нативного человеческого 17-1А антигена, отличающийся тем, что содержит человеческую VH цепь и человеческую VL цепь, которые были функционально перегруппированы.
24. Рецептор по п.23, отличающийся тем, что указанный эпитоп содержит аминокислотную последовательность по меньшей мере одного из пептидов №8, 11, 13, 14, 59, 60, 77 и 79, как определено в таблице 3.
25. Рецептор по п.24, отличающийся тем, что VH цепь имеет одну из двух аминокислотных последовательностей, описанных в SEQ ID No 143 (аминокислотные положения со 108 по 234) и SEQ ID No 145 (аминокислотные положения со 139 по 251), и/или VL цепь имеет одну из двух аминокислотных последовательностей, описанных в SEQ ID No 141 (аминокислотные положения с 1 по 107) и SEQ ID No 147 (аминокислотные положения со 125 по 231).
26. Фармацевтическая композиция, способная конъюгировать антигены в человеческом организме, содержащая рецептор по любому из пп.16-21 и/или рецептор для человеческого антигена по любому из пп.23-25 и фармацевтически приемлемый носитель.
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| Application Number | Priority Date | Filing Date | Title |
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| EP97106109 | 1997-04-14 | ||
| EP97106109.8 | 1997-04-14 |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| RU2552161C2 (ru) * | 2007-07-12 | 2015-06-10 | Пьер Фабр Медикамент | Новые антитела, ингибирующие димеризацию с-мет, и их применения |
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| EP1982990A1 (en) | 1998-03-19 | 2008-10-22 | Human Genome Sciences, Inc. | Cytokine receptor common gamma chain like |
| US7157418B1 (en) | 1998-07-22 | 2007-01-02 | Osprey Pharmaceuticals, Ltd. | Methods and compositions for treating secondary tissue damage and other inflammatory conditions and disorders |
| JP2002537769A (ja) | 1999-02-26 | 2002-11-12 | ヒューマン ジノーム サイエンシーズ， インコーポレイテッド | ヒトエンドカインαおよび使用方法 |
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| US20030031675A1 (en) | 2000-06-06 | 2003-02-13 | Mikesell Glen E. | B7-related nucleic acids and polypeptides useful for immunomodulation |
| CA2413160A1 (en) | 2000-06-15 | 2001-12-20 | Human Genome Sciences, Inc. | Human tumor necrosis factor delta and epsilon |
| MXPA02012434A (es) | 2000-06-16 | 2004-09-06 | Cambridge Antibody Tech | Anticuerpos que se unen inmunoespecificamente a estimulador de linfocitos ii. |
| TWI327600B (en) | 2000-11-28 | 2010-07-21 | Medimmune Llc | Methods of administering/dosing anti-rsv antibodies for prophylaxis and treatment |
| DK1355919T3 (da) | 2000-12-12 | 2011-03-14 | Medimmune Llc | Molekyler med længere halveringstider, sammensætninger og anvendelser deraf |
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| RU2552161C2 (ru) * | 2007-07-12 | 2015-06-10 | Пьер Фабр Медикамент | Новые антитела, ингибирующие димеризацию с-мет, и их применения |
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