Monoclonal Antibody Against NRP-1 b1b2

Abstract

Neuropilin-1 is a member of the neuropilins family protein, which contains a large extracellular domain (a1a2, b1b2 and c), a single transmembrane domain, and a short cytoplasmic tail. NRP-1 plays a critical role in angiogenesis and stimulates endothelial cell division and migration by binding VEGF(165) with b1b2 domain. In the present study, we report the establishment of a monoclonal antibody (A6-26-11-26 clone) specific for NRP-1 b1b2 through hybridoma method. Western blot analysis indicates that our NRP-1 b1b2 MAb can combine both NRP-1 b1b2 and NRP-1 originated from tumor cells. This monoclonal antibody against NRP-1 b1b2 will be useful in the further development of cancer target strategy.

A monoclonal antibody targeting neuropilin-1 inhibits adhesion of MCF7 breast cancer cells to fibronectin by suppressing the FAK/p130cas signaling pathway

“…Briefly, Balb/c mice were injected with hybridoma cells and ascites, and anti-NRP-1 mAbs were obtained after 10 days. mAbs were purified by rProtein A Sepharose column chromatography [46] and diluted in PBS. The purity and concentration of anti-NRP-1 mAbs were assessed by 8% SDS-PAGE gel and Bradford assay, respectively.…”

“…Western blot results from our previous study showed that anti-NRP-1 mAb was specifically combined with both NRP-1 b1b2 recombinant protein and whole NPR-1 [46]. To identify the purity of the current anti-NRP-1 mAb obtained from ascites, anti-NRP-1 mAb was resolved by 8% SDS-PAGE gel (Fig.…”

A Monoclonal Antibody Produced Against Naked2

“…All cell colonies were selected and their supernatants were screened for antibody titers against NKD2 1-217 by indirect ELISA. Three cell colonies with high titer were chosen and cloned by the limiting dilution method, (11,12) and the colony with the highest titer was transferred into 75 cm 2 tissue culture flasks with RPMI-1640 medium containing 10% fetal bovine serum. The hybridoma culture supernatants were then collected for isotyping analysis.…”

Anti‑neuropilin‑1 monoclonal antibody suppresses the migration and invasion of human gastric cancer cells via Akt dephosphorylation

“…Due to the involvement of NRP-1 in the development of malignant tumors and potential advantages of anti-NRP-1 mAbs as a cancer therapy, studies into novel anti-NRP-1 mAbs with greater specificity are warranted. Previous studies by our group have identified an mAb (A6-26-11-26 clone) against the b1/b2 domains of NRP-1 (abbreviation: anti-NRP-1 mAb) ( 22 , 42 , 43 ), first discovered by Li et al ( 42 ), who employed a hybridoma method to screen for b1/b2-specific mAbs. Subsequent analysis by western blotting indicated that the anti-NRP-1 mAb may combine with recombinant human NRP-1-b1/b2 protein fragments and whole NRP-1 proteins expressed by tumor cells ( 42 ).…”