[Preparation and identification of monoclonal antibody against human Flt-1]

Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2012 May;28(5):524-8.
[Article in Chinese]

Abstract

Aim: To prepare functional monoclonal antibodies(mAb)against recombinant human Flt-1(rhFlt-1).

Methods: 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.

Results: 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.

Conclusion: 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.

Publication types

  • English Abstract
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antibodies, Monoclonal / immunology*
  • Cell Line
  • Enzyme-Linked Immunosorbent Assay
  • Female
  • Humans
  • Hybridomas / immunology
  • K562 Cells
  • Mice
  • Mice, Inbred BALB C
  • Vascular Endothelial Growth Factor A / genetics
  • Vascular Endothelial Growth Factor Receptor-1 / analysis
  • Vascular Endothelial Growth Factor Receptor-1 / immunology*

Substances

  • Antibodies, Monoclonal
  • Vascular Endothelial Growth Factor A
  • FLT1 protein, human
  • Vascular Endothelial Growth Factor Receptor-1