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
QDs完成签到,获得积分20
1秒前
2秒前
潘治辉应助研友_Z3NGvn采纳,获得10
2秒前
李爱国应助之风百度采纳,获得10
3秒前
3秒前
Ecrho完成签到,获得积分10
3秒前
5秒前
5秒前
5秒前
起风了发布了新的文献求助10
6秒前
6秒前
7秒前
MQ&FF发布了新的文献求助10
7秒前
7秒前
淳于汲完成签到,获得积分20
8秒前
852应助2miiin采纳,获得10
9秒前
tuotuo完成签到 ,获得积分10
9秒前
充电宝应助yyyyy5采纳,获得10
10秒前
香蕉觅云应助yyyyy5采纳,获得10
10秒前
听风发布了新的文献求助10
11秒前
淳于汲发布了新的文献求助10
12秒前
狂奔的蜗牛完成签到 ,获得积分10
12秒前
12秒前
奋斗的雅柔应助躬身入局采纳,获得10
13秒前
14秒前
雨季发布了新的文献求助10
14秒前
CodeCraft应助胖Q采纳,获得10
14秒前
小二郎应助芥9采纳,获得10
15秒前
大个应助zhscu采纳,获得10
15秒前
JYXin发布了新的文献求助10
17秒前
兴奋纸鹤应助霍千易采纳,获得30
21秒前
24秒前
24秒前
明亮的安波完成签到,获得积分10
24秒前
25秒前
25秒前
天天快乐应助immunity1983采纳,获得10
26秒前
完美世界应助起风了采纳,获得10
29秒前
zhscu发布了新的文献求助10
29秒前
30秒前
高分求助中
Evolution 3rd edition 1500
保险藏宝图 1000
Lire en communiste 1000
Mantiden: Faszinierende Lauerjäger Faszinierende Lauerjäger 700
PraxisRatgeber: Mantiden: Faszinierende Lauerjäger 700
Mathematics and Finite Element Discretizations of Incompressible Navier—Stokes Flows 500
A new species of Coccus (Homoptera: Coccoidea) from Malawi 500
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 3183276
求助须知:如何正确求助?哪些是违规求助? 2833432
关于积分的说明 7993965
捐赠科研通 2495516
什么是DOI,文献DOI怎么找? 1331557
科研通“疑难数据库(出版商)”最低求助积分说明 636352
邀请新用户注册赠送积分活动 603509