Construction of human Alzheimer's disease specific single chain antibodies library
10.3969/j.issn.1009-0126.2015.11.020
- VernacularTitle:人源性阿尔茨海默病噬菌体单链抗体库的构建
- Author:
Nan LI
;
Jianping WANG
- Publication Type:Journal Article
- Keywords:
amyloid beta-protein;
Alzheimer disease;
bacteriophages;
antibodies,monoclonal
- From:
Chinese Journal of Geriatric Heart Brain and Vessel Diseases
2015;(11):1197-1200
- CountryChina
- Language:Chinese
-
Abstract:
Objective To construct the human Alzheimer's disease (AD) specific single chain anti‐bodies (scFv) library for sceening human AD scFv to Aβ1‐42 oligomers .Methods RNA was isola‐ted from 40 ml peripheral blood taken from 18 AD patients .Variable heavy (V H ) and variable light (VL ) genes were amplified by RT‐PCR and linked to the scFv fragments that were then cloned into the phage vector pCANTAB5E .The scFv library was constructed by electroporating E .coli TG1 cells into the pCANTAB5E and rescuing the assistant phage M13K07 .Results The total RNA was extracted .The VH and VL genes were amplified .Electropharesis showed that the length of VH and VL genes was 360bp and 300bp respectively and the length of linked scFv was 750bp .The 2 .4 × 109 scFv library was thus constructed and identified by BstN I digestion .Elec‐tropharesis showed that the length of BstN I‐digested scFv fragments varied ,indicating that the library is of a good diversity .Conclusion The human AD phage scFv library we constructed lays a foundation for screening the antibodies to Aβ1‐42 oligomers and the treatment of AD .
