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
绿萝完成签到,获得积分10
1秒前
pcr163应助lokelnai67采纳,获得80
2秒前
2秒前
朱之欣完成签到,获得积分10
3秒前
默顿的笔记本完成签到,获得积分20
3秒前
uu发布了新的文献求助10
4秒前
浊人发布了新的文献求助10
5秒前
9秒前
9秒前
wwyy完成签到,获得积分10
10秒前
10秒前
Peng应助yan采纳,获得10
11秒前
11秒前
11秒前
懵懂的冰夏完成签到,获得积分20
11秒前
12秒前
13秒前
LiAlan发布了新的文献求助10
13秒前
wwe完成签到,获得积分10
14秒前
15秒前
cc7jn发布了新的文献求助10
15秒前
Eita完成签到,获得积分10
16秒前
戒骄戒躁发布了新的文献求助10
16秒前
李健应助善良的人采纳,获得10
19秒前
uu完成签到,获得积分10
19秒前
20秒前
奉里外完成签到,获得积分10
22秒前
纯情的夜阑完成签到,获得积分10
25秒前
奉里外发布了新的文献求助20
25秒前
要努力发光完成签到,获得积分20
29秒前
HHHAN完成签到,获得积分10
29秒前
30秒前
魔幻海豚完成签到 ,获得积分10
32秒前
梅仑西西发布了新的文献求助10
33秒前
xc发布了新的文献求助10
34秒前
35秒前
Owen应助懵懂的冰夏采纳,获得30
38秒前
39秒前
xc完成签到,获得积分10
41秒前
我不爱池鱼应助阿腾采纳,获得10
43秒前
高分求助中
Spray / Wall-interaction Modelling by Dimensionless Data Analysis 2000
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
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 3184620
求助须知:如何正确求助?哪些是违规求助? 2834922
关于积分的说明 8002218
捐赠科研通 2497295
什么是DOI,文献DOI怎么找? 1332783
科研通“疑难数据库(出版商)”最低求助积分说明 636685
邀请新用户注册赠送积分活动 604062