[Preparation of monoclonal antibodies against hFGF-21 and identification of epitope].
OBJECTIVE
To prepare monoclonal antibodies (mAbs) against human fibroblast growth factor 21 (hFGF-21), and identify the epitope of hFGF-21 mAb through bacterial display.


METHODS
With hFGF-21 as an immunogen and detective antigen, we screened hybridoma cell lines secreting anti-hFGF-21 mAbs using indirect ELISA. The different fragments of hFGF-21 were cloned into bacterial display vector (Apex) to identify the epitope by fluorescence-activated cell sorting (FACS) using FITC-labeled mAbs.


RESULTS
We obtained a stable hybridoma cell line secreting mAbs against hFGF-21; the heavy and light chains of the mAb were IgG 2b and Kappa, respectively. The ascites titer of the hybridoma cell line was 1:4.096×10(6);. The cell line was stable after 30 passages or when stored in liquid nitrogen for 3 months. Western blotting showed the mAbs could bind to hFGF-21 specifically, and could cross-react with murine FGF-21. FACS indicated that this antibody could bind to downstream 107-121 amino acids of hFGF-21.


CONCLUSIONS
The mAbs against hFGF-21 we prepared showed high specificity and stability; the epitope of the mAbs was 107-121 amino acids of hFGF-21.
Department of Biopharmaceutics, Northeast Agricultural University, Harbin, China. haozhichao12345@hotmail.com
Zhichao
Z
Hao
Liming
L
Xu
Yin
Y
Bai
Qi
Q
Wang
Wenfei
W
Wang
Deshan
D
Li
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000737:chemistry / Q000276:immunology*; D002460:Cell Line; D000939:Epitopes / Q000032:analysis / Q000276:immunology*; D005260:Female; D005346:Fibroblast Growth Factors / Q000276:immunology*; D006801:Humans; D006825:Hybridomas / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C
D000911:Antibodies, Monoclonal; D000939:Epitopes; C414620:fibroblast growth factor 21; D005346:Fibroblast Growth Factors