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Generation of Recombinant Human IgG Monoclonal Antibodies from Immortalized Sorted B Cells
Published on: June 5, 2015
Improved fusion partners transfected with DNA fragment encoding IL-11 on generation of human B lymphocyte hybridomas
1Department of Immunology, Fourth Military Medical University, Shaanxi, P.R. China. zhuyong@ihw.com.cn
IL-11, a less identified cytokine, possesses some overlapping functions with IL-6 that are able to facilitate the growth and antibody secretion of B lymphocyte hybridomas. In this report, a DNA fragment encoding human IL-11 was transduced into fusion partners (mouse myelomas Ag8.653 and SP2/0, and human lymphoblastoid cell line HF2) mediated by lipofection. The transfected cells selected with G418 secreted IL-11 constitutively over the range of 32.4 +/- 10.5 units/ml to 76.6 +/- 18.4 units/ml, which could be inhibited by an IL-11 neutralizing MAb up to 80%. The fusion frequency of PBMC doubled, while that of LCLs displayed a 2.4- or 3.3-fold increase, when fused with the transfected fusion partners, respectively. The derived hybridomas from IL-11 secreting fusion partner secreted 3 or 4 times as many immunoglobulins as that from its ancestor. Our data indicate that IL-11 gene transfected fusion partners are improved cell lines for generation of human B lymphocyte hybridomas, and IL-11 may contribute to the increased fusion frequency and antibody secretion of B lymphocyte hybridomas.
IL-11, a less identified cytokine, possesses some overlapping functions with IL-6 that are able to facilitate the growth and antibody secretion of B lymphocyte hybridomas. In this report, a DNA fragment encoding human IL-11 was transduced into fusion partners (mouse myelomas Ag8.653 and SP2/0, and human lymphoblastoid cell line HF2) mediated by lipofection. The transfected cells selected with G418 secreted IL-11 constitutively over the range of 32.4 +/- 10.5 units/ml to 76.6 +/- 18.4 units/ml, which could be inhibited by an IL-11 neutralizing MAb up to 80%. The fusion frequency of PBMC doubled, while that of LCLs displayed a 2.4- or 3.3-fold increase, when fused with the transfected fusion partners, respectively. The derived hybridomas from IL-11 secreting fusion partner secreted 3 or 4 times as many immunoglobulins as that from its ancestor. Our data indicate that IL-11 gene transfected fusion partners are improved cell lines for generation of human B lymphocyte hybridomas, and IL-11 may contribute to the increased fusion frequency and antibody secretion of B lymphocyte hybridomas.
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