RU2014128324A - Антигенсвязывающая молекула, способная к многократному связыванию двух или более молекул антигена - Google Patents
Антигенсвязывающая молекула, способная к многократному связыванию двух или более молекул антигена Download PDFInfo
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- histidine
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- 239000000427 antigen Substances 0.000 title claims abstract 34
- 108091007433 antigens Proteins 0.000 title claims abstract 34
- 102000036639 antigens Human genes 0.000 title claims abstract 34
- 230000027455 binding Effects 0.000 title claims abstract 21
- HNDVDQJCIGZPNO-UHFFFAOYSA-N histidine Natural products OC(=O)C(N)CC1=CN=CN1 HNDVDQJCIGZPNO-UHFFFAOYSA-N 0.000 claims abstract 18
- 238000000034 method Methods 0.000 claims abstract 16
- 238000001727 in vivo Methods 0.000 claims abstract 6
- 238000003780 insertion Methods 0.000 claims abstract 6
- 230000037431 insertion Effects 0.000 claims abstract 6
- 150000001413 amino acids Chemical class 0.000 claims abstract 5
- 238000004519 manufacturing process Methods 0.000 claims abstract 4
- 101710181478 Envelope glycoprotein GP350 Proteins 0.000 claims abstract 2
- 230000003042 antagnostic effect Effects 0.000 claims abstract 2
- 108090000623 proteins and genes Proteins 0.000 claims 9
- 238000010494 dissociation reaction Methods 0.000 claims 1
- 230000005593 dissociations Effects 0.000 claims 1
- 125000000487 histidyl group Chemical group [H]N([H])C(C(=O)O*)C([H])([H])C1=C([H])N([H])C([H])=N1 0.000 claims 1
- 238000006467 substitution reaction Methods 0.000 claims 1
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Abstract
1. Способ улучшения фармакокинетики антитела, включающий замену, по крайней мере, одной аминокислоты CDR указанного антитела гистидином или вставку, по крайней мере, одного гистидина в CDR указанного антитела, где указанные замена гистидином или его вставка увеличивают значение KD(pH 5,8)/KD(pH 7,4) по сравнению со значением KD(pH 5,8)/KD(pH 7,4) до указанных замены гистидином или его вставки, где значение KD(pH 5,8)/KD(pH 7,4) определяется как соотношение KD для антигена при pH 5,8 и KD для антигена при pH 7,4.2. Способ увеличения числа раз связывания с антигеном для антитела in vivo, включающий замену, по крайней мере, одной аминокислоты CDR указанного антитела гистидином или вставку, по крайней мере, одного гистидина в CDR указанного антитела, где указанные замена гистидином или его вставка увеличивают значение KD(pH 5,8)/KD(pH 7,4) по сравнению со значением KD(pH 5,8)/KD(pH 7,4) до указанных замены гистидином или его вставки, где значение KD(pH 5,8)/KD(pH 7,4) определяется как соотношение KD для антигена при pH 5,8 и KD для антигена при pH 7,4.3. Способ по любому из пп. 1 и 2, где значение KD(pH 5,8)/KD(pH 7,4), определяемое как соотношение между KD для антигена при рН 5,8 и KD для антигена при pH 7,4, составляет 2 или больше.4. Способ по любому из пп. 1 и 2, где значение KD(pH 5,8)/KD(pH 7,4) составляет 10 или больше.5. Способ по любому из пп. 1 и 2, в котором значение KD(pH 5,8)/KD(pH 7,4) составляет 40 или больше.6. Способ по любому из пп. 1 и 2, где указанное антитело обладает антагонистической активностью.7. Способ по любому из пп. 1 и 2, где указанное антитело связывается с мембранным антигеном или растворимым антигеном.8. Способ получения антитела, который включает стадии:(a) определения антигенсвязывающей активности антитела при pH от 6,7 до 10,0;(b) определения антигенсвязывающей активности антитела при pH от 4,0 до 6,5;(c) отбора антитела, антигенсвязывающая активность которого при pH от 6,7 до 10,0 выше антигенсвязыв
Claims (12)
1. Способ улучшения фармакокинетики антитела, включающий замену, по крайней мере, одной аминокислоты CDR указанного антитела гистидином или вставку, по крайней мере, одного гистидина в CDR указанного антитела, где указанные замена гистидином или его вставка увеличивают значение KD(pH 5,8)/KD(pH 7,4) по сравнению со значением KD(pH 5,8)/KD(pH 7,4) до указанных замены гистидином или его вставки, где значение KD(pH 5,8)/KD(pH 7,4) определяется как соотношение KD для антигена при pH 5,8 и KD для антигена при pH 7,4.
2. Способ увеличения числа раз связывания с антигеном для антитела in vivo, включающий замену, по крайней мере, одной аминокислоты CDR указанного антитела гистидином или вставку, по крайней мере, одного гистидина в CDR указанного антитела, где указанные замена гистидином или его вставка увеличивают значение KD(pH 5,8)/KD(pH 7,4) по сравнению со значением KD(pH 5,8)/KD(pH 7,4) до указанных замены гистидином или его вставки, где значение KD(pH 5,8)/KD(pH 7,4) определяется как соотношение KD для антигена при pH 5,8 и KD для антигена при pH 7,4.
3. Способ по любому из пп. 1 и 2, где значение KD(pH 5,8)/KD(pH 7,4), определяемое как соотношение между KD для антигена при рН 5,8 и KD для антигена при pH 7,4, составляет 2 или больше.
4. Способ по любому из пп. 1 и 2, где значение KD(pH 5,8)/KD(pH 7,4) составляет 10 или больше.
5. Способ по любому из пп. 1 и 2, в котором значение KD(pH 5,8)/KD(pH 7,4) составляет 40 или больше.
6. Способ по любому из пп. 1 и 2, где указанное антитело обладает антагонистической активностью.
7. Способ по любому из пп. 1 и 2, где указанное антитело связывается с мембранным антигеном или растворимым антигеном.
8. Способ получения антитела, который включает стадии:
(a) определения антигенсвязывающей активности антитела при pH от 6,7 до 10,0;
(b) определения антигенсвязывающей активности антитела при pH от 4,0 до 6,5;
(c) отбора антитела, антигенсвязывающая активность которого при pH от 6,7 до 10,0 выше антигенсвязывающей активности при pH от 4,0 до pH 6,5;
(d) получения гена, кодирующего антитело, отобранное на стадии (c); и
(e) продуцирования антитела, используя полученный на стадии (d) ген, и
где
(1) фармакокинетика указанного антитела является улучшенной; или
(2) число раз связывания с антигеном для указанного антитела является увеличеным in vivo.
9. Способ получения антитела, который включает стадии:
(a) связывания антитела с антигеном при параметре pH, составляющем от 6,7 до 10,0;
(b) допущения нахождения антитела, связанного с антигеном на стадии (a), при параметре рН, составляющем от 4,0 до 6,5;
(c) сбора антитела, которое подвергалось диссоциации при параметре pH, составляющем от 4,0 до 6,5;
(d) получения гена, кодирующего антитело, полученное на стадии (c); и
(e) продуцирования антитела, используя полученный на стадии (d) ген, и
где
(1) фармакокинетика указанного антитела является улучшенной; или
(2) число раз связывания с антигеном для указанного антитела является увеличенным in vivo.
10. Способ получения антитела, активность связывания которого при рН от 6,7 до 10,0 превышает активность связывания при рН 4,0 до 6,5, который включает стадии:
(a) связывания антитела с колонкой с иммобилизованным антигеном при параметре рН от 6,7 до 10,0;
(b) элюирования антитела, которое связалось с колонкой при рН от 6,7 до 10,0, с колонки при параметре pH 4,0 до 6,5;
(c) сбора подвергшегося элюции антитела;
(d) получения гена, кодирующего антитела, полученного на стадии (c); и
(e) продуцирования антитела, используя полученный на стадии (d) ген, и
где
(1) фармакокинетика указанного антитела является улучшенной; или
(2) число раз связывания с антигеном для указанного антитела является увеличенным in vivo.
11. Способ получения антитела, активность связывания которого при рН от 6,7 до 10,0 превышает активность связывания при рН 4,0 до 6,5, который включает стадии:
(a) связывания библиотеки антитела с колонкой с иммобилизованным антигеном при параметре рН от 6,7 до 10,0;
(b) элюирования антитела с колонки при параметре pH от 4,0 до 6,5;
(c) амплификации гена, кодирующего элюируемое антитело;
(d) сбора подвергшегося элюции антитела;
(e) получения гена, кодирующего антитело, собранное на стадии (d); и
(f) продуцирования антитела, используя полученный на стадии (е) ген, и
где
(1) фармакокинетика указанного антитела является улучшенной; или
(2) число раз связывания с антигеном для указанного антитела является увеличенным in vivo.
12. Способ получения по любому из пп. 7-11, который дополнительно включает стадию замещения, по крайней мере, одной аминокислоты указанного антитела гистидином или вставки, по крайней мере, одного гистидина в указанное антитело.
Applications Claiming Priority (6)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP2008-104147 | 2008-04-11 | ||
JP2008104147 | 2008-04-11 | ||
JP2008247713 | 2008-09-26 | ||
JP2008-247713 | 2008-09-26 | ||
JP2009-068744 | 2009-03-19 | ||
JP2009068744 | 2009-03-19 |
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RU2010145939/10A Division RU2571225C2 (ru) | 2008-04-11 | 2009-04-10 | Антигенсвязывающая молекула, способная к многократному связыванию двух или более молекул антигена |
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