CN107058235A - 一种b细胞筛选方法及其在单克隆抗体制备中的应用 - Google Patents
一种b细胞筛选方法及其在单克隆抗体制备中的应用 Download PDFInfo
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- CN107058235A CN107058235A CN201710417596.9A CN201710417596A CN107058235A CN 107058235 A CN107058235 A CN 107058235A CN 201710417596 A CN201710417596 A CN 201710417596A CN 107058235 A CN107058235 A CN 107058235A
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Cited By (6)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN109517058A (zh) * | 2018-10-24 | 2019-03-26 | 普健生物(武汉)科技有限公司 | 一种筛选能应用于流式细胞术抗体的方法 |
CN110016462A (zh) * | 2019-02-20 | 2019-07-16 | 优睿赛思(武汉)生物科技有限公司 | 从脾脏细胞中高效分离单个抗原特异性b淋巴细胞的方法 |
CN111303294A (zh) * | 2018-12-11 | 2020-06-19 | 澳世科技(深圳)有限公司 | 一种细胞表面展现式快速筛选抗体表达细胞的方法 |
CN112094810A (zh) * | 2020-08-26 | 2020-12-18 | 中国农业大学 | 单b细胞筛选方法及其在小分子单克隆抗体制备中的应用 |
CN114651065A (zh) * | 2019-09-03 | 2022-06-21 | 五松尖端医疗产业振兴财团 | 导入用于提高蛋白质生产性的单个条形码***的信号肽超高速筛选方法 |
CN115166241A (zh) * | 2022-08-22 | 2022-10-11 | 广东忠信生物科技有限公司 | 一种同时筛选记忆b细胞和浆细胞的高效筛选技术及应用 |
Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO1990000625A1 (en) * | 1988-07-14 | 1990-01-25 | Monoclonetics International, Inc. | Pre-screening for depletion and enrichment of specific b-cells |
CN103603057A (zh) * | 2013-07-03 | 2014-02-26 | 深圳大学 | 基于可感染病毒颗粒型抗体重链库/轻链库的抗体筛选和制备方法 |
WO2014146074A2 (en) * | 2013-03-15 | 2014-09-18 | Alder Biopharmaceuticals, Inc. | Protocol for identifying and isolating antigen-specific b cells and producing antibodies to desired antigens |
-
2017
- 2017-06-05 CN CN201710417596.9A patent/CN107058235B/zh active Active
Patent Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO1990000625A1 (en) * | 1988-07-14 | 1990-01-25 | Monoclonetics International, Inc. | Pre-screening for depletion and enrichment of specific b-cells |
WO2014146074A2 (en) * | 2013-03-15 | 2014-09-18 | Alder Biopharmaceuticals, Inc. | Protocol for identifying and isolating antigen-specific b cells and producing antibodies to desired antigens |
CN103603057A (zh) * | 2013-07-03 | 2014-02-26 | 深圳大学 | 基于可感染病毒颗粒型抗体重链库/轻链库的抗体筛选和制备方法 |
Non-Patent Citations (2)
Title |
---|
VILASACK THAMMAVONGSA ET AL: "Staphylococcal manipulation of host immune responses", 《NAT REV MICROBIOL.》 * |
李节等: "金黄色葡萄球菌蛋白A(Staphylococcal Protein A,SPA)分泌型融合表达***", 《生物工程进展》 * |
Cited By (9)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN109517058A (zh) * | 2018-10-24 | 2019-03-26 | 普健生物(武汉)科技有限公司 | 一种筛选能应用于流式细胞术抗体的方法 |
CN111303294A (zh) * | 2018-12-11 | 2020-06-19 | 澳世科技(深圳)有限公司 | 一种细胞表面展现式快速筛选抗体表达细胞的方法 |
CN110016462A (zh) * | 2019-02-20 | 2019-07-16 | 优睿赛思(武汉)生物科技有限公司 | 从脾脏细胞中高效分离单个抗原特异性b淋巴细胞的方法 |
CN110016462B (zh) * | 2019-02-20 | 2020-04-24 | 优睿赛思(武汉)生物科技有限公司 | 从脾脏细胞中高效分离单个抗原特异性b淋巴细胞的方法 |
CN114651065A (zh) * | 2019-09-03 | 2022-06-21 | 五松尖端医疗产业振兴财团 | 导入用于提高蛋白质生产性的单个条形码***的信号肽超高速筛选方法 |
CN112094810A (zh) * | 2020-08-26 | 2020-12-18 | 中国农业大学 | 单b细胞筛选方法及其在小分子单克隆抗体制备中的应用 |
CN112094810B (zh) * | 2020-08-26 | 2022-07-05 | 中国农业大学 | 单b细胞筛选方法及其在小分子单克隆抗体制备中的应用 |
CN115166241A (zh) * | 2022-08-22 | 2022-10-11 | 广东忠信生物科技有限公司 | 一种同时筛选记忆b细胞和浆细胞的高效筛选技术及应用 |
CN115166241B (zh) * | 2022-08-22 | 2023-03-24 | 广东忠信生物科技有限公司 | 一种同时筛选记忆b细胞和浆细胞的高效筛选技术及应用 |
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