In vitro affinity maturation of human GM-CSF antibodies by targeted CDR-diversification

抗体 亲和力成熟 细胞因子 生物 受体 体外 体细胞突变 免疫系统 免疫学 分子生物学 细胞生物学 B细胞 生物化学
作者
Stefan Steidl,Olaf Ratsch,Bodo Brocks,Manuela Dürr,Elisabeth Thomassen-Wolf
出处
期刊:Molecular Immunology [Elsevier]
卷期号:46 (1): 135-144 被引量:85
标识
DOI:10.1016/j.molimm.2008.07.013
摘要

The mammalian immune system applies somatic hypermutation to select for antibodies with improved dissociation rates in vivo up to an intrinsic limit, previously termed as affinity ceiling. However, for certain therapeutic applications it may be desirable to further improve antibody affinities beyond that limit. In this study the selection of antibodies specific for the pro-inflammatory cytokine granulocyte-macrophage colony-stimulating factor (GM-CSF) from the HuCAL GOLD human antibody library is described. In order to increase affinity and also functional activity, in vitro affinity maturation of a pool of lead Fab candidates was carried out. CDR-L3 and parallel CDR-H2 diversification using trinucleotide consensus cassettes were followed by the combination of optimized CDR-L3 and CDR-H2 leading to a 5000-fold improved affinity finally reaching a KD of 400 fM. Cytokine neutralizing potential of MOR04357 was evaluated in a TF-1 proliferation assay. Along with affinity optimization a 2000-fold increase in potency was observed compared to the parental antibody. Due to species cross-reactivity MOR04357 also blocks rat GM-CSF induced proliferation of FDCP-1 cells. Receptor inhibition studies showed that MOR04357 prevents the interaction of GM-CSF with the GM-CSF receptor alpha chain. As a consequence this leads to a blockade in signal transduction as measured by abolished STAT5 phosphorylation in the presence of GM-CSF and antibody. Due to its pro-inflammatory role GM-CSF has been implicated in the pathophysiology of inflammatory diseases like rheumatoid arthritis or asthma. Based on the mode of action described herein MOR04357 shows favourable antibody features as a potential drug candidate.
最长约 10秒,即可获得该文献文件

科研通智能强力驱动
Strongly Powered by AbleSci AI
更新
大幅提高文件上传限制,最高150M (2024-4-1)

科研通是完全免费的文献互助平台,具备全网最快的应助速度,最高的求助完成率。 对每一个文献求助,科研通都将尽心尽力,给求助人一个满意的交代。
实时播报
kk发布了新的文献求助10
刚刚
lalala完成签到,获得积分10
1秒前
飞雪之舞完成签到,获得积分10
1秒前
orixero应助Raxiny采纳,获得10
1秒前
晴天完成签到,获得积分10
2秒前
SciGPT应助阳宝是个小蜜蜂采纳,获得10
2秒前
muyi完成签到,获得积分10
3秒前
jessie发布了新的文献求助10
3秒前
4秒前
4秒前
4秒前
6秒前
7秒前
激动的乐安完成签到 ,获得积分10
7秒前
善学以致用应助Chem is try采纳,获得10
7秒前
柔弱云朵完成签到,获得积分10
7秒前
8秒前
飞龙爵士发布了新的文献求助10
9秒前
9秒前
爆米花应助落落采纳,获得10
9秒前
归海亦云完成签到,获得积分10
10秒前
10秒前
嘴嘴发布了新的文献求助10
12秒前
姬霓太美发布了新的文献求助10
12秒前
李健的小迷弟应助清璃采纳,获得10
13秒前
华仔应助张德帅采纳,获得10
13秒前
Ava应助元谷雪采纳,获得10
14秒前
15秒前
宋怡慷完成签到,获得积分10
15秒前
15秒前
守护使者发布了新的文献求助10
15秒前
小贝发布了新的文献求助10
15秒前
CodeCraft应助dont_love_study采纳,获得10
16秒前
hyx完成签到,获得积分10
16秒前
16秒前
hyx发布了新的文献求助10
20秒前
ding应助7777采纳,获得10
20秒前
20秒前
不配.应助HJY采纳,获得20
21秒前
21秒前
高分求助中
Spray / Wall-interaction Modelling by Dimensionless Data Analysis 2000
Mixed-anion Compounds 600
ALA生合成不全マウスでの糖代謝異常の分子機構解析 520
Aspects of Babylonian celestial divination: the lunar eclipse tablets of Enūma Anu Enlil 500
Geochemistry, 2nd Edition 地球化学经典教科书第二版 401
2024 Medicinal Chemistry Reviews 400
Dictionary of socialism 350
热门求助领域 (近24小时)
化学 医学 生物 材料科学 工程类 有机化学 生物化学 物理 内科学 纳米技术 计算机科学 化学工程 复合材料 基因 遗传学 催化作用 物理化学 免疫学 量子力学 细胞生物学
热门帖子
关注 科研通微信公众号,转发送积分 3199091
求助须知:如何正确求助?哪些是违规求助? 2847981
关于积分的说明 8065682
捐赠科研通 2512742
什么是DOI,文献DOI怎么找? 1344693
科研通“疑难数据库(出版商)”最低求助积分说明 639863
邀请新用户注册赠送积分活动 609663