RU2014114172A - Легковыделяемые биспецифические антитела с природным иммуноглобулиновым форматом - Google Patents
Легковыделяемые биспецифические антитела с природным иммуноглобулиновым форматом Download PDFInfo
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- 108060003951 Immunoglobulin Proteins 0.000 title claims abstract 5
- 102000018358 immunoglobulin Human genes 0.000 title claims abstract 5
- 238000000034 method Methods 0.000 claims abstract 23
- 102000006496 Immunoglobulin Heavy Chains Human genes 0.000 claims abstract 13
- 108010019476 Immunoglobulin Heavy Chains Proteins 0.000 claims abstract 13
- 102000025171 antigen binding proteins Human genes 0.000 claims abstract 13
- 108091000831 antigen binding proteins Proteins 0.000 claims abstract 13
- 150000007523 nucleic acids Chemical class 0.000 claims abstract 13
- 108020004707 nucleic acids Proteins 0.000 claims abstract 12
- 102000039446 nucleic acids Human genes 0.000 claims abstract 12
- 102000004169 proteins and genes Human genes 0.000 claims abstract 10
- 108090000623 proteins and genes Proteins 0.000 claims abstract 10
- 102000013463 Immunoglobulin Light Chains Human genes 0.000 claims abstract 7
- 108010065825 Immunoglobulin Light Chains Proteins 0.000 claims abstract 7
- 230000004048 modification Effects 0.000 claims abstract 6
- 238000012986 modification Methods 0.000 claims abstract 6
- 238000012258 culturing Methods 0.000 claims abstract 2
- 210000004027 cell Anatomy 0.000 claims 10
- 241000700605 Viruses Species 0.000 claims 4
- 210000004978 chinese hamster ovary cell Anatomy 0.000 claims 2
- 230000002163 immunogen Effects 0.000 claims 2
- 108700024394 Exon Proteins 0.000 claims 1
- 241000238631 Hexapoda Species 0.000 claims 1
- 241000282412 Homo Species 0.000 claims 1
- 238000001042 affinity chromatography Methods 0.000 claims 1
- 239000003607 modifier Substances 0.000 claims 1
- 210000001525 retina Anatomy 0.000 claims 1
- 210000002966 serum Anatomy 0.000 claims 1
- 239000007787 solid Substances 0.000 claims 1
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- C07K1/088—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length using protecting groups or activating agents using activating agents containing other elements, e.g. B, Si, As
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- C07K16/24—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against cytokines, lymphokines or interferons
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- C07K16/24—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against cytokines, lymphokines or interferons
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- 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
- C07K16/2803—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants against the immunoglobulin superfamily
- C07K16/2809—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants against the immunoglobulin superfamily against the T-cell receptor (TcR)-CD3 complex
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- 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
- C07K16/2866—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants against receptors for cytokines, lymphokines, interferons
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- 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
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Abstract
1. Способ получения антигенсвязывающего белка, который является гетеродимерным в отношении связывания белка А, включающий:культивирование клетки, содержащей(i) первую нуклеиновую кислоту, кодирующую первую тяжелую цепь иммуноглобулина, содержащую первый вариабельный домен, который распознает первый эпитоп, причем первая тяжелая цепь иммуноглобулина содержит первую область СН3 IgG человека, выбранную из IgG1, IgG2 и IgG4, где первая область СН3 связывается с белком А;(ii) вторую нуклеиновую кислоту, кодирующую вторую тяжелую цепь иммуноглобулина, содержащую второй вариабельный домен, который распознает второй эпитоп, причем вторая тяжелая цепь иммуноглобулина содержит вторую область СН3 IgG человека, выбранную из IgG1, IgG2 и IgG4, где вторая область СН3 содержит модификацию, которая снижает или предотвращает связывание второй области СН3 с белком А по сравнению с первой областью СН3;(iii) третью нуклеиновую кислоту, кодирующую легкую цепь иммуноглобулина, которая образует пару с первой и второй тяжелой цепью иммуноглобулина;в условиях, обеспечивающих экспрессию клеткой первой и второй тяжелой цепи иммуноглобулина и легкой цепи иммуноглобулина с образованием антигенсвязывающего белка, где антигенсвязывающий белок является гетеродимерным в отношении связывания белка А.2. Способ по п. 1, в котором первая и вторая тяжелые цепи иммуноглобулина представляют собой тяжелые цепи IgG человека.3. Способ по п. 2, в котором каждая из первой и второй тяжелых цепей иммуноглобулина представляет собой тяжелую цепь IgG1 человека.4. Способ по п. 1, в котором легкая цепь иммуноглобулина представляет собой легкую цепь иммуноглобулина человека.5. Спос�
Claims (19)
1. Способ получения антигенсвязывающего белка, который является гетеродимерным в отношении связывания белка А, включающий:
культивирование клетки, содержащей
(i) первую нуклеиновую кислоту, кодирующую первую тяжелую цепь иммуноглобулина, содержащую первый вариабельный домен, который распознает первый эпитоп, причем первая тяжелая цепь иммуноглобулина содержит первую область СН3 IgG человека, выбранную из IgG1, IgG2 и IgG4, где первая область СН3 связывается с белком А;
(ii) вторую нуклеиновую кислоту, кодирующую вторую тяжелую цепь иммуноглобулина, содержащую второй вариабельный домен, который распознает второй эпитоп, причем вторая тяжелая цепь иммуноглобулина содержит вторую область СН3 IgG человека, выбранную из IgG1, IgG2 и IgG4, где вторая область СН3 содержит модификацию, которая снижает или предотвращает связывание второй области СН3 с белком А по сравнению с первой областью СН3;
(iii) третью нуклеиновую кислоту, кодирующую легкую цепь иммуноглобулина, которая образует пару с первой и второй тяжелой цепью иммуноглобулина;
в условиях, обеспечивающих экспрессию клеткой первой и второй тяжелой цепи иммуноглобулина и легкой цепи иммуноглобулина с образованием антигенсвязывающего белка, где антигенсвязывающий белок является гетеродимерным в отношении связывания белка А.
2. Способ по п. 1, в котором первая и вторая тяжелые цепи иммуноглобулина представляют собой тяжелые цепи IgG человека.
3. Способ по п. 2, в котором каждая из первой и второй тяжелых цепей иммуноглобулина представляет собой тяжелую цепь IgG1 человека.
4. Способ по п. 1, в котором легкая цепь иммуноглобулина представляет собой легкую цепь иммуноглобулина человека.
5. Способ по п. 1, в котором модификация выбрана из группы, состоящей из (a) 95R и (b) 95R и 96F в нумерации экзонов IMGT, или (a′) 435R и (b′) 435R и 436F в нумерации системы EU.
6. Способ по п. 5, в котором вторая область СН3 дополнительно содержит от одной до пяти модификаций, выбранных из группы, состоящей из 16Е, 18М, 44S, 52N, 57М и 821 в системе нумерации экзонов IMGT или 356Е, 358М, 384S, 392N, 397М и 4221 в системе нумерации EU.
7. Способ по п. 1, в котором модификация не изменяет времени полужизни в сыворотке антигенсвязывающего белка по сравнению с соответствующим антигенсвязывающим белком, не имеющим модификации.
8. Способ по п. 5, в котором вторая область СН3 антигенсвязывающего белка является неиммуногенной или по существу неиммуногенной для человека.
9. Способ по п. 1, в котором клетка выбрана из клетки СНО, клетки COS, клетки 293, клетки HeLa, клетки насекомого и клетки сетчатой оболочки глаза.
10. Способ по п. 9, в котором клетка представляет собой клетку СНО.
11. Способ по п. 1, дополнительно включающий выделение антигенсвязывающего белка с использованием белка А.
12. Способ по п. 11, в котором антигенсвязывающий белок выделяют на колонке для аффинной хроматографии с белком А на твердом носителе, используя градиент pH и ионный модификатор.
13. Способ по п. 12, в котором градиент pH представляет собой ступенчатый градиент, включающий одну или несколько ступеней pH между значениями pH, равными 3 и 5.
14. Способ по п. 13, в котором антигенсвязывающий белок элюируется при значениях pH между приблизительно 3,9 и приблизительно 4,4.
15. Способ по п. 14, в котором антигенсвязывающий белок элюируется при значении pH приблизительно 4,2.
16. Способ по п. 1, в котором каждая из первой и второй нуклеиновой кислоты получены из генетически модифицированной мыши.
17. Способ по п. 16, в котором генетически модифицированная мышь содержит одну или несколько последовательностей нуклеиновых кислот вариабельной области тяжелой цепи человека.
18. Способ по п. 1, в котором первая, вторая и третья нуклеиновые кислоты введены в клетку с использованием вектора или вируса, несущего эти нуклеиновые кислоты.
19. Способ по п. 18, в котором:
(a) последовательности первой и второй нуклеиновых кислот находятся на одном и том же векторе или вирусе;
(b) последовательности первой и третьей нуклеиновых кислот находятся на одном и том же векторе или вирусе; или
(c) последовательности первой, второй и третьей нуклеиновых кислот находятся на одном и том же векторе или вирусе.
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| US22068709P | 2009-06-26 | 2009-06-26 | |
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| RU2012102602/10A Division RU2522002C2 (ru) | 2009-06-26 | 2010-06-25 | Легковыделяемые биспецифические антитела с природным иммуноглобулиновым форматом |
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| RU2014114172/10A RU2569157C2 (ru) | 2009-06-26 | 2014-04-10 | Легковыделяемые биспецифические антитела с природным иммуноглобулиновым форматом |
| RU2015140911A RU2647758C2 (ru) | 2009-06-26 | 2015-09-25 | Легковыделяемые биспецифические антитела с природным иммуноглобулиновым форматом |
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| EP (3) | EP2975051B1 (ru) |
| JP (6) | JP5816170B2 (ru) |
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