基于高通量表面等离子体共振技术筛选与TPBG具有高亲和力的单克隆抗体研究
Screening study of monoclonal antibodies with high affinity to TPBG based on high-throughput surface plasmon resonance technology
江秀玲 1王晓丽 1胡有根 2彭建明 1严慧深1
作者信息
- 1. 扬州市职业大学医学院,江苏 扬州 225009
- 2. 武警江苏总队医院,江苏 扬州 225009
- 折叠
摘要
目的 探讨从制备的单克隆抗体中快速筛选出与滋养层糖蛋白(TPBG)具有高亲和力的单克隆抗体的方法.方法 采用高通量表面等离子体共振技术(SPR),将不同种属的TPBG通过氨基偶联的方式固定在GLC传感器芯片上,梯度稀释的抗体作为流通相,筛选能高度亲和TPBG的单克隆抗体,并测定相互作用的动力学常数.抗体1结合人和食蟹猴TPBG,亲和力分别为65.0、31.6 nmol/L;抗体6结合人、小鼠、食蟹猴TPBG,亲和力分别为77.6、92.1、87.7 nmol/L.结果 该研究成功筛选出2种能与TPBG特异性结合的单克隆抗体.动力学图谱显示,这2种抗体与TPBG蛋白的特异性结合稳定.结论 采用SPR可筛选出与TP-BG蛋白具有高亲和力的单克隆抗体,为开发出一些新型的抗肿瘤药物提供一种方法.
Abstract
Objective To investigate a method of rapidly screen monoclonal antibodies with high affini-ty to trophoblastic glycoprotein(TPBG)from the prepared monoclonal antibodies.Methods High-through-put surface plasmon resonance(SPR)technology was used to immobilize TPBG of different species on the GLC sensor chip by amino coupling.Gradient dilution of the antibody was used as the flow phase to screen monoclonal antibodies that can highly affinity TPBG,and the kinetic constants of the interaction were deter-mined.Antibody 1 binds to human and cynomolgus monkey TPBG with affinity of 65.0 and 31.6 nmol/L,re-spectively.Antibody 6 binds to human,mouse and cynomolgus monkey TPBG with affinity of 77.6,92.1 and 87.7 nmol/L,respectively.Results This study successfully screened two monoclonal antibodies that can spe-cifically bind to TPBG.Meanwhile,the specific binding of these two antibodies to TPBG protein was stable by binding kinetic.Conclusion The monoclonal antibodies with high affinity to TPBG protein are screened out by SPR,which provides a method for the development of some new anti-tumor drugs.
关键词
5T4蛋白/高通量/表面等离子体共振/亲和力/单克隆抗体/抗肿瘤药物Key words
TPBG protein/High-throughput/Surface plasmon resonance/Affinity/Mono-clonal antibodies/Anti-tumor drug引用本文复制引用
基金项目
江苏省自然科学基金面上项目(18KJD36003)
扬州市职业大学校级自然科学基金项目(2018ZR28)
出版年
2024
国家科技期刊平台
NSTL
万方数据