Synthesis of human Fab

人类Fab的合成

• 批准号: 14406020
• 负责人: YAMASHIRO Tetsu
• 金额: $ 3.46万
• 依托单位: Oita University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2002
• 资助国家: 日本
• 起止时间: 2002 至 2003
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-14406020/
• 关键词: combinatorial science; phage display; dengue infection; tropical medicine; infectious diseases; humanized antibody; Antibody engineering; antibody therapy; ファージディスプレイ; 抗体; デングウイルス; 抗体療法; 分子生物学

The sero-prevalence of dengue virus (DEN)-specific IgG was determined among 1008 adults and among 201 children younger than 10 years old, in Santo Domingo Dominican Republic Most adults (98%, 987/1008) and 56% (113/201) of children were positive for DEN-specific IgG. Among children, the prevalence of DEN-specific IgG increased with age, that is, 1 to 2 yr, 0--5%, 3 to 6 yr, 25 --65%, and 【greater than or equal】 7 yr, 76 --92%. Fifty percent of the prevalence shown in <1 group was probably due to maternal antibody transfer. Neutralization tests of adult samples indicate that DEN infections with different serotypes have occurred sequentially or simultaneously among the inhabitants of Santo Domingo. Fab monoclonal antibodies to DEN4 were recovered from mRNA from immune higher primate. Two Fabs neutralized DEN4 efficiently with a titer of 0.24-0.48 micro g/mL by PRNT. One of the Fab was converted to full-length IgG1 by combining it with human sequences. The humanized IgG1 neutralized DEN4 strains from different geographical origins at a similar PRNT50 titer. A human Fab library was constructed from PBL of volunteers who had recovered from severe DEN infection. The phage library was panned several times with DEN1, DEN2, DEN3, and DEN4 virions, and recombinant DEN4 NS1 protein, and Japanese encephalitis virus virion.

对多米尼加共和国圣多明各1008名成人和201名10岁以下儿童进行登革病毒（DEN）特异性IgG血清流行率测定，大多数成人（98%,987/1008）和56%（113/201）的儿童登革病毒（DEN）特异性IgG阳性。在儿童中，den特异性IgG的患病率随着年龄的增长而增加，即1 - 2岁、0- 5%、3 -6岁、25 -65%和【大于或等于】7岁，76 -92%。<1组50%的患病率可能是由于母体抗体转移所致。成人样本的中和试验表明，不同血清型的DEN感染先后或同时在圣多明各居民中发生。从免疫高等灵长类动物的mRNA中分离到DEN4的Fab单克隆抗体。两个fab通过PRNT有效中和DEN4，滴度为0.24-0.48微g/mL。其中一个Fab通过与人类序列结合转化为全长IgG1。人源化IgG1能以相似的PRNT50滴度中和来自不同地理来源的DEN4菌株。从重度DEN感染恢复的志愿者PBL中构建人类Fab文库。用DEN1、DEN2、DEN3和DEN4病毒粒子、重组DEN4 NS1蛋白和乙型脑炎病毒粒子对噬菌体文库进行多次筛选。

项目成果

T.Yamashiro: "Synthesis of useful antibody molecules using the combinatorial library and the phage display technologies"Jpn.J.Trop.Med.Hyg.. 31. 24-25 (2003)

T.Yamashiro：“使用组合文库和噬菌体展示技术合成有用的抗体分子”Jpn.J.Trop.Med.Hyg.. 31. 24-25 (2003)

R.Men, T.Yamashiro, et al.: "Identification of chimpanzee Fab fragments by repertoire cloning and production of a full-length humanized IgG1 antibody highly efficient for neutralization of dengue type 4 virus."Journal of virology. (in press). (2004)

R.Men、T.Yamashiro 等人：“通过全长克隆和生产全长人源化 IgG1 抗体来鉴定黑猩猩 Fab 片段，可高效中和登革热 4 型病毒。”病毒学杂志。

T.Yamashiro, et al.: "Seroprevalence of IgG specific to Dengue virus among adults and children of Santo Domingo, Dominican Republic."American Journal of Tropical Medicine and hygiene. (in press). (2004)

T.Yamashiro 等人：“多米尼加共和国圣多明各成人和儿童中登革热病毒特异性 IgG 血清阳性率”。《美国热带医学与卫生杂志》。

R.Men, T.Yamashiro, et al.: "Identification of chimpanzee Fab fragments by repertoire cloning and production of a full-length humanized IgG lantibody highly efficient for neutralization of dengue type 4 virus."Journal of Virology. (in press). (2004)

R.Men、T.Yamashiro 等人：“通过全长克隆和生产全长人源化 IgG 羊毛抗体来鉴定黑猩猩 Fab 片段，可高效中和登革热 4 型病毒。”病毒学杂志。

Men, R., Yamashiro, T.et al.: "Identification of chimpanzee Fab fragments by repertoire cloning and Production of a full-length humanized IgG1 antibody highly efficient for neutralization of"Journal of Virology. (in press).

Men, R., Yamashiro, T.等人：“通过库克隆鉴定黑猩猩 Fab 片段并生产高效中和的全长人源化 IgG1 抗体”《病毒学杂志》。