Expression of Human-Mouse Chimeric Antibody against TNF-α in CHO Cells
HE Tai-ping
Abstract
HE Tai-ping
Abstract
Objective To construct human-mouse chimeric antibody against TNF-α and express in CHO cells.Methods Clone the gene encoding variable region of immunoglobulin from murine hybridoma cells by RT-PCR and RACE methods.Construct human-mouse chimeric antibody by fusing the cloned gene with the gene encoding the constant region of human immunoglobulin.Transfect CHO cells with the fusion gene and detect the expressed product.Results The primary expression level of the constructed chimeric antibody in CHO cells was 0.71 μg/ml,and that after subclone was 0.95 μg/ml.Conclusion Anti-human TNF-α chimeric antibody gene was successfully constructed and highly expressed in CHO cells.
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Objective To construct human-mouse chimeric antibody against TNF-α and express in CHO cells.Methods Clone the gene encoding variable region of immunoglobulin from murine hybridoma cells by RT-PCR and RACE methods.Construct human-mouse chimeric antibody by fusing the cloned gene with the gene encoding the constant region of human immunoglobulin.Transfect CHO cells with the fusion gene and detect the expressed product.Results The primary expression level of the constructed chimeric antibody in CHO cells was 0.71 μg/ml,and that after subclone was 0.95 μg/ml.Conclusion Anti-human TNF-α chimeric antibody gene was successfully constructed and highly expressed in CHO cells.
Key concepts: Chinese hamster ovary cell, Molecular biology, Antibody, clone (Java method), Transfection, Chimeric gene, Fusion gene, Biology