PMID: 22558992May 9, 2012Paper

Preparation and identification of monoclonal antibody against human Flt-1

Xi bao yu fen zi mian yi xue za zhi = Chinese journal of cellular and molecular immunology
Yin-liang XieYuanfu Xu

Abstract

To prepare functional monoclonal antibodies(mAb)against recombinant human Flt-1(rhFlt-1). A cell line stable secreting mAb was established by using FLT-1 extracellular domain III as antigen and hybridoma technique. Then it was purified in large-scale from mouse ascites by protein G affinity chromatography. The characteristics of mAb were then determined by ELISA, Western blotting and FACS. The immunoglobin subtype of mAb XA12 was IgG1 with kappa (κ) light chains, and it could recognize rhFlt-1 specifically. Furthermore, mAb XA12 could bind to rhFlt-1with high affinity (K=1.28±0.05 nmol/L). It could also be used to detect Flt-1-positive cells, such as human umbilical vascular endothelial cells (HUVECs) and K562/A02 in a dose-dependent fashion. A hybridoma cell line secreting functional anti-rhFlt-1 mAb was successfully prepared. The antibody can be used to study the function of Flt-1 and further potentially optimized for clinical purpose.

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