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RU2010148904A - Антитела к интерлейкину-1альфа и способы применения - Google Patents

Антитела к интерлейкину-1альфа и способы применения Download PDF

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RU2010148904A
RU2010148904A RU2010148904/10A RU2010148904A RU2010148904A RU 2010148904 A RU2010148904 A RU 2010148904A RU 2010148904/10 A RU2010148904/10 A RU 2010148904/10A RU 2010148904 A RU2010148904 A RU 2010148904A RU 2010148904 A RU2010148904 A RU 2010148904A
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amino acid
specifically binds
mab
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Джон СИМАРД (US)
Джон СИМАРД
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ИксБиотеч, Инк. (CA)
ИксБиотеч, Инк.
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Abstract

1. Очищенное человеческое IgG1 mAb (моноклональное антитело), которое специфически связывается с человеческим IL-1α, содержащее тяжелую цепь, ковалентно связанную с легкой цепью, где тяжелая цепь содержит аминокислотную последовательность SEQ ID NO: 9, и легкая цепь содержит аминокислотную последовательность SEQ ID NO: 11. ! 2. Набор выделенных нуклеиновых кислот, содержащих первую нуклеиновую кислоту, кодирующую тяжелую цель человеческого IgGI mAb, которое специфически связывается с IL-1α, и вторую нуклеиновую кислоту, кодирующую легкую цепь человеческого IgGI mAb, которое специфически связывается с человеческим IL-1α, где первая нуклеиновая кислота кодирует аминокислотную последовательность SEQ ID NO: 9, и вторая нуклеиновая кислота кодирует аминокислотную последовательность SEQ ID NO: 11. ! 3. Набор выделенных нуклеиновых кислот по п.2, где первая нуклеиновая кислота содержит нуклеотидную последовательность SEQ ID NO: 10, и вторая нуклеиновая кислота содержит нуклеотидную последовательность SEQ ID NO: 12. ! 4. Набор выделенных нуклеиновых кислот по п.2, который содержится по меньшей мере в одном экспрессирующем векторе. ! 5. Набор выделенных нуклеиновых кислот по п.3, который содержится по меньшей мере в одном экспрессирующем векторе. ! 6. Набор выделенных нуклеиновых кислот по п.4, который содержится в выделенной клетке-хозяине. ! 7. Набор выделенных нуклеиновых кислот по п.6, где клетка-хозяин представляет собой клетку млекопитающих. ! 8. Способ уничтожения клетки, экспрессирующей человеческий IL-1α, включающий стадию приведения клетки в контакт с очищенным человеческим IgGI mAb, которое специфически связывается с человеческим IL-1α, где указанная клетка унич

Claims (20)

1. Очищенное человеческое IgG1 mAb (моноклональное антитело), которое специфически связывается с человеческим IL-1α, содержащее тяжелую цепь, ковалентно связанную с легкой цепью, где тяжелая цепь содержит аминокислотную последовательность SEQ ID NO: 9, и легкая цепь содержит аминокислотную последовательность SEQ ID NO: 11.
2. Набор выделенных нуклеиновых кислот, содержащих первую нуклеиновую кислоту, кодирующую тяжелую цель человеческого IgGI mAb, которое специфически связывается с IL-1α, и вторую нуклеиновую кислоту, кодирующую легкую цепь человеческого IgGI mAb, которое специфически связывается с человеческим IL-1α, где первая нуклеиновая кислота кодирует аминокислотную последовательность SEQ ID NO: 9, и вторая нуклеиновая кислота кодирует аминокислотную последовательность SEQ ID NO: 11.
3. Набор выделенных нуклеиновых кислот по п.2, где первая нуклеиновая кислота содержит нуклеотидную последовательность SEQ ID NO: 10, и вторая нуклеиновая кислота содержит нуклеотидную последовательность SEQ ID NO: 12.
4. Набор выделенных нуклеиновых кислот по п.2, который содержится по меньшей мере в одном экспрессирующем векторе.
5. Набор выделенных нуклеиновых кислот по п.3, который содержится по меньшей мере в одном экспрессирующем векторе.
6. Набор выделенных нуклеиновых кислот по п.4, который содержится в выделенной клетке-хозяине.
7. Набор выделенных нуклеиновых кислот по п.6, где клетка-хозяин представляет собой клетку млекопитающих.
8. Способ уничтожения клетки, экспрессирующей человеческий IL-1α, включающий стадию приведения клетки в контакт с очищенным человеческим IgGI mAb, которое специфически связывается с человеческим IL-1α, где указанная клетка уничтожается.
9. Способ по п.8, где человеческое IgGI mAb содержит тяжелую цепь, содержащую аминокислотную последовательность SEQ ID NO: 9, и легкую цепь, содержащую аминокислотную последовательность SEQ ID NO: 11.
10. Способ ингибирования миграции клетки через матрикс базальной мембраны, включающий стадию добавления очищенного mAb, которое специфически связывается с человеческим IL-1α, к смеси, содержащей матрикс базальной мембраны и указанную клетку.
11. Способ по п.10, где очищенное mAb представляет собой человеческое IgGI mAb, содержащее тяжелую цепь, ковалентно связанную с легкой цепью.
12. Способ по п.11, где тяжелая цепь содержит аминокислотную последовательность SEQ ID NO: 9, и легкая цепь содержит аминокислотную последовательность SEQ ID NO: 11.
13. Способ ингибирования IL-1α-индуцированного увеличения экспрессии Е-селектина на поверхности человеческой эндотелиальной клетки, включающий стадию добавления очищенного mAb, которое специфически связывается с человеческим IL-1α, к смеси, содержащей эндотелиальную клетку и IL-1α, где количество Е-селектина на поверхности клетки уменьшается.
14. Способ по п.13, где очищенное mAb представляют собой человеческое IgGI mAb, содержащее тяжелую цепь, ковалентно связанную с легкой цепью.
15. Способ по п.14, где тяжелая цепь содержит аминокислотную последовательность SEQ ID NO: 9, и легкая цепь содержит аминокислотную последовательность SEQ ID NO: 11.
16. Способ контролирования воспаления у субъекта-человека, ранее подвергнутого стадиям получения от субъекта первого образца мононуклеарных клеток периферической крови в первый раз; приведения первого образца в контакт с очищенным mAb, которое специфически связывается с человеческим IL-1α; и определения в первом образце процента клеток, которые связываются с моноклональным Аb, включающий стадии:
(а) получение от субъекта второго образца мононуклеарных клеток периферической крови во второй раз;
(б) приведение второго образца в контакт с очищенным mAb, которое специфически связывается с человеческим IL-1α;
(в) определение во втором образце процента клеток, которые связываются с моноклональным Аb; и
(г) сравнение процента клеток, которые связываются с mAb, в первом образце с процентом клеток, которые связываются с моноклональным Аb, во втором образце.
17. Способ по п.16, где очищенное mAb представляют собой человеческое IgGI mAb, содержащее тяжелую цепь, ковалентно связанную с легкой цепью.
18. Способ по п.17, где тяжелая цепь содержит аминокислотную последовательность SEQ ID NO: 9, и легкая цепь содержит аминокислотную последовательность SEQ ID NO: 11.
19. Способ, включающий стадии:
(а) обогащение биологического образца, полученного от субъекта-человека с использованием фильтра, который разделяет молекулы в соответствии с молекулярной массой на первую фракцию, содержащую интактный IgG, образующий комплекс с IL-1α, и вторую фракцию, содержащую молекулы менее 100 кДа; и
(б) определение количества IL-1α в первой фракции.
20. Способ по п.19, включающий стадии:
(а) обогащение образца плазмы, полученной от субъекта-человека с использованием фильтра, который разделяет молекулы в соответствии с молекулярной массой на первую фракцию, содержащую интактный IgG, образующий комплекс с IL-1α, и вторую фракцию, содержащую молекулы менее 100 кДа;
(б) добавление первой фракции к субстрату, содержащему иммобилизованные Abs против IgG человека, в условиях, которые позволяют IgG в первой фракции специфически связываться с Abs против IgG человека, иммобилизованными на субстрате;
(в) промывание субстрата для удаления из первой фракции вещества, которое не связывается специфически с иммобилизованными Abs против IgG человека;
(г) приведение субстрата, промытого на стадии (в), в контакт с АЬ, которое специфически связывается с человеческим IL-1α, в условиях, которые позволяют АЬ, которое специфически связывается с человеческим IL-1α, специфически связываться с любым человеческим IL-1α, связанными с указанным субстратом;
(д) промывание субстрата для удаления любого Аb, которое специфически связывается с человеческим IL-1α, который не связан с субстратом; и
(е) определение количества Аb, которое специфически связывается с человеческим IL-1α, оставшегося связанным с субстратом после стадии (д).
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