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
Rosebud发布了新的文献求助10
1秒前
ZhouKL发布了新的文献求助10
4秒前
4秒前
40873完成签到,获得积分10
5秒前
5秒前
晗月完成签到,获得积分10
6秒前
打打应助QI采纳,获得10
6秒前
8秒前
嗯嗯哈哈发布了新的文献求助10
8秒前
8秒前
大寒给大寒的求助进行了留言
8秒前
wanci应助韦远侵采纳,获得10
8秒前
汉堡包应助小篆采纳,获得10
9秒前
黄小北完成签到,获得积分10
9秒前
QI完成签到,获得积分10
11秒前
所所应助AAA采纳,获得10
11秒前
11秒前
houyunfeng完成签到 ,获得积分10
11秒前
Ava应助阳光的芯采纳,获得10
12秒前
英俊的铭应助wrufhg采纳,获得30
13秒前
王太白完成签到,获得积分10
14秒前
小鱼发布了新的文献求助10
14秒前
打打应助强度采纳,获得10
17秒前
追寻的问安完成签到,获得积分10
18秒前
18秒前
yiyiyiyiyi//完成签到 ,获得积分10
19秒前
19秒前
19秒前
Rosebud完成签到,获得积分10
20秒前
20秒前
orixero应助聪明的远锋采纳,获得10
21秒前
ah完成签到,获得积分10
21秒前
111完成签到,获得积分10
22秒前
22秒前
HHW发布了新的文献求助10
23秒前
qq关闭了qq文献求助
23秒前
诩阽完成签到,获得积分10
23秒前
暮霭沉沉应助包包采纳,获得10
23秒前
uiwh发布了新的文献求助30
24秒前
畅快白亦发布了新的文献求助10
24秒前
高分求助中
Spray / Wall-interaction Modelling by Dimensionless Data Analysis 2000
Aspects of Babylonian celestial divination: the lunar eclipse tablets of Enūma Anu Enlil 500
Mathematics and Finite Element Discretizations of Incompressible Navier—Stokes Flows 500
Dictionary of socialism 350
Mixed-anion Compounds 300
Geochemistry, 2nd Edition 地球化学经典教科书第二版 300
Idoxuridine 260
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 3195624
求助须知:如何正确求助?哪些是违规求助? 2844459
关于积分的说明 8050116
捐赠科研通 2509095
什么是DOI,文献DOI怎么找? 1341427
科研通“疑难数据库(出版商)”最低求助积分说明 639124
邀请新用户注册赠送积分活动 608325