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
1秒前
curtisness完成签到,获得积分0
1秒前
1秒前
2秒前
3秒前
fissh发布了新的文献求助10
3秒前
hbzyydx46发布了新的文献求助30
4秒前
1234发布了新的文献求助10
4秒前
02完成签到,获得积分10
4秒前
rr_完成签到,获得积分10
4秒前
nicelily完成签到 ,获得积分20
4秒前
成就紫真发布了新的文献求助10
5秒前
Liuuhhua完成签到,获得积分10
5秒前
NB发布了新的文献求助10
7秒前
YY完成签到 ,获得积分10
7秒前
仙人殊恍惚应助研友_LN7x6n采纳,获得10
8秒前
8秒前
8秒前
啊奶糖发布了新的文献求助10
9秒前
和谐乌冬面完成签到 ,获得积分10
9秒前
yufanhui应助忧郁秋尽采纳,获得10
9秒前
许钟一完成签到,获得积分10
10秒前
于鱼完成签到,获得积分10
10秒前
缪飞珍完成签到,获得积分10
11秒前
biofresh完成签到,获得积分10
11秒前
12秒前
sanyiwen完成签到,获得积分20
12秒前
可爱的函函应助自由觅波采纳,获得10
13秒前
快乐的冰棍完成签到,获得积分10
13秒前
神勇的香露完成签到,获得积分10
14秒前
名丿完成签到,获得积分10
14秒前
june1111完成签到,获得积分10
15秒前
15秒前
安详伯云完成签到,获得积分20
15秒前
sanyiwen发布了新的文献求助10
16秒前
sarah完成签到,获得积分10
17秒前
fissh完成签到 ,获得积分20
17秒前
mini完成签到,获得积分10
19秒前
19秒前
19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 3184696
求助须知:如何正确求助?哪些是违规求助? 2834971
关于积分的说明 8002491
捐赠科研通 2497387
什么是DOI,文献DOI怎么找? 1332822
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 604068