Production of antibody fragments against urokinase by Bacillus brevis and its application
短芽孢杆菌生产抗尿激酶抗体片段及其应用
基本信息
- 批准号:06556015
- 负责人:
- 金额:$ 5.25万
- 依托单位:
- 依托单位国家:日本
- 项目类别:Grant-in-Aid for Developmental Scientific Research (B)
- 财政年份:1994
- 资助国家:日本
- 起止时间:1994 至 1995
- 项目状态:已结题
- 来源:
- 关键词:
项目摘要
Expression-secretion vectors for the production of anti-human urokinase antibody fragments, single chain (sc) Fv, Fab', and scFab', were constructed and inroduced into Bacillus brevis. The promoter of the cell wall protein gene operon (cwp), and the translation start site and the signal pepetide-encoding region from the mwp gene, the gene for one of the major cell wall proteins of B.brevis, were used in the vector. The cDNAs encoding the light and heavy chains of Fv and Fab' fragments were prepared by PCR from the cDNAs for mouse-human chimeric monoclonal antibody against urokinase-type plasminogen activator and inserted immediately downstream of the signal peptide-encoding region. To construct the genes for sc fragments, the cDNAs encoding the light and heavy chains were linked by synthetic oligonucleotides encoding flexible peptide linkers of 17 or 24 amino acids. Fab' was efficiently produced by B.brevis resulting in an accumulation at a level of 100 mg/I in the culture medium. In contrast, the amounts of scFv and scFab' produced by B.brevis were at a level of a few mg/I in the culture medium. Fab' produced by B.brevis was purified and confirmed to have an antigen binding activity comparable to that of the parental antibody.
构建了单链(sc) Fv、Fab’和scFab’表达-分泌载体,并将其引入短芽孢杆菌中。该载体利用短螺旋体细胞壁蛋白基因操纵子(cwp)启动子、短螺旋体细胞壁蛋白基因mwp的翻译起始位点和信号肽编码区。将小鼠-人抗尿激酶型纤溶酶原激活物嵌合单克隆抗体的cdna通过PCR制备Fv和Fab’片段的轻链和重链cdna,并直接插入信号肽编码区下游。为了构建sc片段的基因,编码轻链和重链的cdna通过合成编码17或24个氨基酸的柔性肽连接体的寡核苷酸连接。短芽孢杆菌能有效地产生Fab',在培养基中积累量为100 mg/I。相比之下,短芽孢杆菌在培养基中产生的scFv和scFab'的数量在几mg/I的水平。经纯化后,该菌株的Fab'具有与亲本抗体相当的抗原结合活性。
项目成果
期刊论文数量(19)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)
Y.Sagiya: "Direct high level secretion into the culture medium of tuna growth hormone in biologically active form by Bacillus brevis." Appl.Microbiol.Biotechnol.42. 358-363 (1994)
Y.Sagiya:“短芽孢杆菌以生物活性形式将金枪鱼生长激素直接高水平分泌到培养基中。”
- DOI:
- 发表时间:
- 期刊:
- 影响因子:0
- 作者:
- 通讯作者:
Tada,Hiroko: "Expression and charatervization of a chimeric bispecific antigody against fibrin and against urokinase-type plasminogen actiator" Journal of Biotechnology. 33. 157-174 (1994)
Tada,Hiroko:“抗纤维蛋白和尿激酶型纤溶酶原激活剂的嵌合双特异性抗体的表达和表征”生物技术杂志。
- DOI:
- 发表时间:
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- 影响因子:0
- 作者:
- 通讯作者:
Sagiya, Yoji: "Direct high-leuel secretion of tuna growth hormone in biologically active furm by Bacillus brevis" Applied microbiology and Biotechnology. 42. 358-363 (1994)
Sagiya,Yoji:“短芽孢杆菌在生物活性菌中直接高水平分泌金枪鱼生长激素”应用微生物学和生物技术。
- DOI:
- 发表时间:
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- 影响因子:0
- 作者:
- 通讯作者:
T.Kurokawa, A.Watanabe, A.Kakinuma, and S.Iwasa: "Thrombolytic properties of an immunoconjugate between single-chain urokinase-type plasminogen activator (scu-PA) and F (ab') _2 of bispecific monoclonal antibody against fibrin and against u-PA in hamsters
T.Kurokawa、A.Watanabe、A.Kakinuma 和 S.Iwasa:“单链尿激酶型纤溶酶原激活剂 (scu-PA) 和抗纤维蛋白双特异性单克隆抗体 F (ab) _2 之间免疫缀合物的溶栓特性
- DOI:
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- 影响因子:0
- 作者:
- 通讯作者:
T.Ishihara: "Cloning and characterization of the gene for a protein thiol-disulfide oxidoreductase in Bacillus brevis." J.Bacteriol.177. 745-749 (1995)
T.Ishihara:“短芽孢杆菌中蛋白质硫醇二硫化物氧化还原酶基因的克隆和表征。”
- DOI:
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- 影响因子:0
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YAMAGATA Hideo其他文献
データサイエンス入門(10)回帰と教師付き分類:多値予測問題
数据科学概论(十)回归与监督分类:多值预测问题
- DOI:
- 发表时间:
2020 - 期刊:
- 影响因子:0
- 作者:
日本軍縮学会(編);阿部達也;妹尾哲志;園田茂人;YAMAGATA Hideo;デービス・コーリン,橋本賢一;須田守;佐藤安信;石綿はる美;椿 広計 - 通讯作者:
椿 広計
国際司法裁判所における反訴の受理可能性
国际法院反诉的可受理性
- DOI:
- 发表时间:
2004 - 期刊:
- 影响因子:0
- 作者:
YAMAGATA;Hideo;YAMAGATA Hideo;山形英郎;山形英郎 - 通讯作者:
山形英郎
割当国籍論の可能性と限界(特集:国籍選択の逆説)
配额国籍理论的可能性与局限性(专题:国籍选择悖论)
- DOI:
- 发表时间:
2018 - 期刊:
- 影响因子:0
- 作者:
Nishi;Makoto;Tetsuya YOKOHAMA;大屋 雄裕;YAMAGATA Hideo;渡辺徹也;利根川佳子;大屋雄裕 - 通讯作者:
大屋雄裕
Le mouvement en faveur de la réforme du droit de
喜爱拉尔运动
- DOI:
- 发表时间:
2017 - 期刊:
- 影响因子:0
- 作者:
日本軍縮学会(編);阿部達也;妹尾哲志;園田茂人;YAMAGATA Hideo;デービス・コーリン,橋本賢一;須田守;佐藤安信;石綿はる美 - 通讯作者:
石綿はる美
Empowerment of Refugees
赋予难民权力
- DOI:
- 发表时间:
2016 - 期刊:
- 影响因子:0
- 作者:
日本軍縮学会(編);阿部達也;妹尾哲志;園田茂人;YAMAGATA Hideo;デービス・コーリン,橋本賢一;須田守;佐藤安信 - 通讯作者:
佐藤安信
YAMAGATA Hideo的其他文献
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{{ truncateString('YAMAGATA Hideo', 18)}}的其他基金
A Study of the Exclusive Defense Doctrine in Article 9 of the Japanese Constitution: New Security Policy beyond the Right of Self-Defense
日本宪法第九条专属防卫主义研究:超越自卫权的新安全政策
- 批准号:
18H00794 - 财政年份:2018
- 资助金额:
$ 5.25万 - 项目类别:
Grant-in-Aid for Scientific Research (B)
Molecular mechanisms of morphogenesis and response to environmental cues in cladoceran crustaceans
枝角类甲壳动物形态发生和对环境信号响应的分子机制
- 批准号:
18570093 - 财政年份:2006
- 资助金额:
$ 5.25万 - 项目类别:
Grant-in-Aid for Scientific Research (C)
Unilateral Use of Force in International Law: A Study on Just War Theories in Modern Age
国际法中的单方面使用武力:现代正义战争理论研究
- 批准号:
16530031 - 财政年份:2004
- 资助金额:
$ 5.25万 - 项目类别:
Grant-in-Aid for Scientific Research (C)
POPULATION ECOLOGY OF ABNORMAL GEMALES PRODUCING ONLY FEMALE PROGENY IN THE GYPSYMOTH
舞毒蛾中仅产雌性后代的异常双翅蛾的种群生态学
- 批准号:
12440220 - 财政年份:2000
- 资助金额:
$ 5.25万 - 项目类别:
Grant-in-Aid for Scientific Research (B)
A systematic analysis of the effects of fine-structures of antibodies on microbial expression and secretion and construction of a general-purpose system for antibody production.
系统分析抗体精细结构对微生物表达和分泌的影响,构建通用抗体生产系统。
- 批准号:
09556019 - 财政年份:1997
- 资助金额:
$ 5.25万 - 项目类别:
Grant-in-Aid for Scientific Research (B).
Molecular mechanism of evolution and morphological diversity in Cladocera.
枝角类进化和形态多样性的分子机制。
- 批准号:
09839034 - 财政年份:1997
- 资助金额:
$ 5.25万 - 项目类别:
Grant-in-Aid for Scientific Research (C)
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