The Screening of Human Anti-Hepatoma Fd Fragments Guided with the Chemeric Light Chain of HAb18
- VernacularTitle:嵌合轻链为模板导向筛选人源性抗肝癌抗体重链Fd片段
- Author:
Guoqiang BAO
;
Qingjiu MA
;
Jinliang XING
;
Xiangmin YANG
;
Zhinan CHEN
- Publication Type:Journal Article
- Keywords:
phage antibody library;
hepatoma;
guided-selection
- From:
Chinese Journal of Cancer Biotherapy
1995;0(03):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective: To select anti-HAb18G (hepatoma associated antigen) human Fd fragments with guided selection of L chain of chimeric Fab-HAb18. Methods: The human Fd repertories genes were amplified by RT-PCR from PBMC of hepatoma patients, and cloned into the vector pComb3X with chimeric L chain to construct the human-mouse hybrid Fab phage library. HAblSGE, extracellular region of HAblSG, was used as antigen to screen. The positive clones was obtained by ELISA and FCM with p Ⅲ-fusion Fab antibody. The DNA sequences were analyzed. Results: A human-mouse Fab antibody library were constructed with 2?107 PFU. After 6 rounds panning, 7 positive clones were obtained with ELISA and FCM. And sequences of 2 clones with better affinity were same. The CHI belonged to the IgG2 class as the parent Fd, and the variable region belonged to VH3 family. Conclusions: Through the construction of the HuMFab phage antibody library and chemeric L chain-guided selection, we get the available human Fd fragments for subsequent research.
