CREATION OF HYBRIDOMA CELL LINES SECRETING MONOCLONAL ANTIBODIES AGAINST NS1 ANTIGEN OF FOUR DENGUE SEROTYPES

Pham Hung Tien, Hoàng Xuân Cường, Thị Bích Thủy Võ, Trịnh Thế Sơn, Đỗ Như Bình

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Abstract

Objectives: To create hybridoma cell lines that secretes monoclonal antibodies against NS1 antigen for 4 types of Dengue virus in BALB/c mice. Methods: Immunize BALB/c mice with recombinant NS1 antigen with 4 strains; collect B lymphocytes and fuse with Myeloma Sp2/0 cells to create a hybridoma cell line capable of producing monoclonal antibodies against NS1; screen the positive hybrid cell line by ELISA reaction. Results: The concentration of NS1 antigen injected into mice was 100 µg/mL in sterile 1x PBS solution, mixed with FCA and FIA adjuvant in 1:1 volume. Fusion of B lymphocytes and Myeloma Sp2/0 cells were conducted in the ratio 1:5, cultured in HAT and HT medium. The percentage of hybrid cells that developed after 3 days accounted for 91.15% - 92.92%. By ELISA reaction, 2 hybrid cell lines 1A2 and 2A1 were selected with the highest OD (Optical Density) values of 1.0324 and 1.3765. Conclusion: Successfully created 2 hybrid cell lines 1A2 and 2A1 producing monoclonal antibodies specific to the recombinant NS1 antigen pooling 4 strains of Dengue virus.

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References

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