Preparation, Purification, and Identification of a Monoclonal Antibody Against NRP2 b1b2 Domain.
First identified as a high-affinity kinase-deficient receptor for class-3 semaphorins and vascular endothelial growth factor (VEGF) families, Neuropilin2 (NRP2) is a transmembrane non-tyrosine-kinase glycoprotein that has a vital function in neuronal patterning. Furthermore, NRP2 expression is often upregulated in cancer tissues and correlated with poor prognosis. In the present study, we report the establishment of a monoclonal antibody specific for NRP2b1b2 domain (NRP2 MAb) through hybridoma method. NRP2 MAb is measured to have a titer of 5.12 × 10(5) against NRP2b1b2 in indirect ELISA. Western blotting, flow cytometry, and immunofluorescence analysis indicate that NRP2 MAb can combine full-length NRP2 in LoVo and SW480 cells. Besides helping further understand NRP2-related pathological mechanisms and cell-signaling pathways, NRP2 MAb may act as a therapeutic agent for cancer in the future.
Cancer Research Center, Medical College, Xiamen University , Xiamen, Fujian, China .
Yun
Y
Yang
Cancer Research Center, Medical College, Xiamen University , Xiamen, Fujian, China .
Na
N
Chen
Cancer Research Center, Medical College, Xiamen University , Xiamen, Fujian, China .
Zhe
Z
Li
Cancer Research Center, Medical College, Xiamen University , Xiamen, Fujian, China .
Xian-Jiang
XJ
Wang
Cancer Research Center, Medical College, Xiamen University , Xiamen, Fujian, China .
Sheng-Yu
SY
Wang
Cancer Research Center, Medical College, Xiamen University , Xiamen, Fujian, China .
Tingwu
Cancer Research Center, Medical College, Xiamen University , Xiamen, Fujian, China .
Fang-Hong
FH
Luo
Cancer Research Center, Medical College, Xiamen University , Xiamen, Fujian, China .
Jiang-Hua
JH
Yan
D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry* / Q000276:immunology*; D002478:Cells, Cultured; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D005260:Female; D005455:Fluorescent Antibody Technique / Q000379:methods; D006825:Hybridomas / Q000276:immunology; D008565:Membrane Proteins / Q000737:chemistry / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D039943:Neuropilin-2 / Q000276:immunology*; D017434:Protein Structure, Tertiary; D015398:Signal Transduction / Q000276:immunology; D015854:Up-Regulation / Q000276:immunology; D042461:Vascular Endothelial Growth Factor A / Q000276:immunology
D000911:Antibodies, Monoclonal; D008565:Membrane Proteins; D039943:Neuropilin-2; D042461:Vascular Endothelial Growth Factor A