Preparation Of Monoclonal Antibody Of Mouse Anti-Human CXCR3-B And Application Of Anti-Tumor Cells

CXCR3 molecule belongs to G-protein receptor family.The length of it is 415 amino acids.It is the only member in CXC with the ELR sequence in N-end.For its relation to hyperplasia of vessel,it is inferred that ELR sequence is related to the function of CXCR3-B.Thus the research on the 52 amino acids in N-end can have effect on the function of CXC in hyperplasia of vessel.It can also explain the way that the PF4 and ELF to repress the growth of cancer.This experiment is making the anti-CXCR3-R molecule antibody and the mechanism of PF4 of repressing the growth of cell.First,make the first 1-19 17-35 and 33-51 amino acids.Meanwhile clone the CXCR-3 gene by using RT-PCR and PCR.If the sequencing is right,purify the GST-CXCR3-B protein.Cut the other part with Factor Xa.Get the 1-51 amino acid in N-end of CXCR3-B molecule.Test the effect and get the clone to merge the cells in order to get the anti-CXCR3-B N-end antibody.After western blotting,do research on the mechanism of CXCR3-B in repressing cancer.The result is that the purity is 98% 88.54%and 80%.After the merging cell the SDS-PAGE shows that target protein expressed.The western-blotting shows that the anti CXCR3-B can identify GST-CXCR3-B merged protein.This result shows that the protein expressed correctly.After the cutting of factor Xa,the western blotting shows that CXCR3-R can be identified by anti-CXCR3-R polyclonal antibody.It can be used in isolating the monoclonal antibody of CXCR3-B.9 CXCR3-B monoclonal antibody were get,and 3 of them are the antibody of the 1-19 amino acid in the N-end of CXCR3-B.2 of them are antibody of the 33-51 amino acids in the N-end of CXCR3-B.3 of them can identify of the 1-51 amino acids in the N-end.And 1 can identify both 1-19 amino acids in the N-end and 33-51 amino acids in the N-end. And the monoclonal antibody can identify the CXCR3-B in tissues.Primary results show that anti-CXCR3-B can prevent PF4 from combining with U937,consequently it can prevent the increase and motion of the cell endothelial cell.