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. 2014 Dec 1;33(6):401–408. doi: 10.1089/mab.2014.0043

FIG. 4.

FIG. 4.

Cloning of monoclonal antibody 3E10 variable regions and production of chimeric antibody. (A) IgH, IgK, and Igλ V regions of CREPT monoclonal antibody 3E10 were amplified from hybridoma cells of 3E10. Water was used as a negative control. (B) Variable sequences of CREPT monoclonal antibody Ig heavy and light chain from 3E10 clone of hybridoma cells. Framework regions are marked with green color and complementarity determining regions are marked with yellow. (C) ELISA assays were performed using different dilutions of CREPT monoclonal antibody 3E10 ascites from mouse hybridoma cells as a primary antibody. An irrelevant antibody was used as negative control. (D) ELISA assays were performed with different dilutions of CREPT monoclonal antibody 3E10 produced in supernatant from HEK293T cells expressing the chimeric antibody. The supernatant from HEK293T cells transfected vector was used as negative control. (E) Western blot analysis was performed in colon cancer samples. CREPT monoclonal antibody 3E10 produced in mouse ascites and supernatant produced by 293T cells were used as primary antibodies to detect colon tumor tissues. P, paired non-tumor tissue; T, tumor tissue from same patient.