Monoclonal Antibody Which Specifically Recognizes B Cell Lymphoma and Use Thereof
a monoclonal antibody and lymphoma technology, applied in the field of monoclonal antibodies, can solve the problems of low complete cure effect of less than 10% of malignant b cells and rituximab, high possibility of side effects, and low effect of rituximab, and achieve excellent anticancer effect, selective cancer cells, and reduced expression of hla
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example 1
Antibodies and Reagents
[0114]The anti-CD74 monoclonal antibody MB741, anti-HLA-DR-FITC, anti-CD19-FITC, anti-CD3 monoclonal antibody, and anti-mouse IgG-FITC & PE were purchased from BD Phamingen, the anti-CD74 monoclonal antibody By2 was purchased from Santa cruz, and the alexa Fluor 546 anti-mouse IgG was purchased from Invitrogen.
[0115]Additionally, recombinant human IL-2 was purchased from Peprotech. RAG2− / −c− / − mice were provided by Central Institute for Experimental Animals (CIEA, Japan). Hela-CIITA (class II transactivator) cell line was provided by Dr. Philippe Pierre (Centre d'Immunologie de Marseille-Luminy, Marseille, France).
example 2
Cell Line
[0116]L3055 cells were cultured in a medium prepared by adding 10% FBS, 3 mM glutamine, 100 U / mL penicillin G, and 100 μg / mL streptomycin to Iscoves modified Dulbeccos medium (IMDM, Irvine Science, Santa Ana, Calif.). THP-1, 293, PC3, Jurkat, CCRF-CEM, A431, 1A2, JVM2, and BC-1 cell lines were purchased from ATCC cell line bank (Manassas, Va.), and SNU638, SNU20, and SNU538 cell lines were provided by the Korean Cell Line Bank (KCLB, Seoul, Korea). EBV-transformed lymphoblastoid cells (LCL) were prepared by infecting peripheral blood mononuclear cells (PBMCs) with a EBV B95-8 cell line, and then culturing in an RPMI1640 medium (WellGene, Korea), which was prepared by adding 10% FBS and antibiotics in a state provided with 1 μg / mL CsA.
example 3
Construction of B Cell Lymphoma-Specific Monoclonal Antibody
[0117]A balb / c mouse was intraperitoneally injected with 2×107 of L3055 cell line a total of two times every two weeks. Three weeks after the second injection, 1×107 of the same cells were intravenously injected. On the 4th day from the final immune injection, the spleen cells of the mice were separated and fused using SP2 / 0 myeloma cells and PEG. The fused cells were aliquoted into a 96-well plate at a concentration of 5×105 cells / well, added with a HAT selective medium containing hypoxanthine (H) and aminopterin, and cultured thereafter.
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