title: [Preparation and identification of monoclonal antibody against human Flt-1].
abstract: OBJECTIVE
  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.
  
  
  CONCLUSIONS
  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.
authors.affiliation: Chinese Academy of Medical Sciences, Tianjin, China.
authors.forename: Yin-liang
authors.initials: YL
authors.lastname: Xie
authors.forename: Li
authors.initials: L
authors.lastname: Ma
authors.forename: Li
authors.initials: L
authors.lastname: Qin
authors.forename: Peng
authors.initials: P
authors.lastname: Liu
authors.forename: Shan-nan
authors.initials: SN
authors.lastname: Cao
authors.forename: Hai-yan
authors.initials: HY
authors.lastname: Zhu
authors.forename: Dong-sheng
authors.initials: DS
authors.lastname: Xiong
authors.forename: Yuan-fu
authors.initials: YF
authors.lastname: Xu
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D002460:Cell Line; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D006801:Humans; D006825:Hybridomas / Q000276:immunology; D020014:K562 Cells; D051379:Mice; D008807:Mice, Inbred BALB C; D042461:Vascular Endothelial Growth Factor A / Q000235:genetics; D040281:Vascular Endothelial Growth Factor Receptor-1 / Q000032:analysis / Q000276:immunology*
chemical_list: D000911:Antibodies, Monoclonal; D042461:Vascular Endothelial Growth Factor A; C501162:FLT1 protein, human; D040281:Vascular Endothelial Growth Factor Receptor-1