Mobile website
The Screening of Human Anti-Hepatoma Fd Fragments Guided with the Chemeric Light Chain of HAb18
- Article
- Figures
- Metrics
- Preview PDF
- Reference
- Related
- Cited by
- Materials
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. HAb18GE, extracellular region of HAb18G, was used as antigen to screen. The positive clones was obtained by ELISA and FCM with pⅢ-fusion Fab antibody. The DNA sequences were analyzed. Results: A human-mouse Fab antibody library were constructed with 2×10 7 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 CH1 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.
Keywords:
Project Supported:
Clc Number:
