Mouse Anti-CEA Recombinant Antibody (clone mAb 8019) (CAT#: FAMAB-0054CQ)

Figure 1 Relative binding of scFv proteins by ELISA.

Plates were coated with mAb 8019 and binding of the scFv was determined by reaction with nickel-nitrilotriacetic acid-alkaline phosphatase conjugate. Results with 10 μg of purified renatured protein per well are shown. Controls without scFv or 8019 were negligible.

Tripathi, P. K., Qin, H., Deng, S., Xu, C., Bhattacharya-Chatterjee, M., Foon, K. A., & Chatterjee, S. K. (1998). Antigen mimicry by an anti-idiotypic antibody single chain variable fragment. Molecular immunology, 35(13), 853-863.

Figure 2 Inhibition of binding of anti-CEA mAb to CEA by purified, renatured 3H1 derived scFv.

Plates were coated with CEA and binding of ¹²⁵I-8019 in the absence and presence of increasing concentrations of intact anti-idiotype antibody and scFv proteins was determined. As control, 11D10 an isotype matched, unrelated intact anti-idiotype antibody was used.

Tripathi, P. K., Qin, H., Deng, S., Xu, C., Bhattacharya-Chatterjee, M., Foon, K. A., & Chatterjee, S. K. (1998). Antigen mimicry by an anti-idiotypic antibody single chain variable fragment. Molecular immunology, 35(13), 853-863.

Figure 3 Immune flow cytometric analysis of sera from mice immunized with 3H1 derived scFv, VH-Ln-VL.

Groups of mice (3:group) were immunized with 25 μg of naked VH-Ln-VL (scFv) or VH-Ln-VL-KLH conjugate (scFv-KLH). Pre-immune sera and the sera after the 4th immunization were pooled and reacted with CEA-negative MC-38 (A-E) and CEA-positive MC-38cea (F-J) cells. Reaction with PBS (A, F); diluted pre-immune sera (B, G) anti-CEA mAb, 8019; (C, H); sera from mice immunized with scFv-KLH (D, I) and naked scFv (E, J) were performed.

Tripathi, P. K., Qin, H., Deng, S., Xu, C., Bhattacharya-Chatterjee, M., Foon, K. A., & Chatterjee, S. K. (1998). Antigen mimicry by an anti-idiotypic antibody single chain variable fragment. Molecular immunology, 35(13), 853-863.

Figure 4 Inhibition of Ab2 (3H1) binding to Ab1 (8019) by monkey (PRO 667) Ab3 sera by radioimmunoassay.

Purified 8019 was used to coat the plate (250 ng/well), and the binding of radiolabeled3H1(~50000cpm) to 8019 was tested for inhibition in the presence of difference dilution of Ab3 sera. In a parallel control expriment, an unrelated Ab1-Ab2 system (mAb BrE1-11D10) was used as the control.

Chakraborty, M., Foon, K. A., Kohler, H., & Bhattacharya-Chatterjee, M. (1995). Preclinical evaluation in nonhuman primates of an anti-idiotypic antibody that mimicks the carcinoembryonic antigen. Journal of immunotherapy with emphasis on tumor immunology: official journal of the Society for Biological Therapy, 18(2), 95-103.

Figure 5 Immunoperoxidase staining of colonic adenocarcinoma and normal colon by monkey Ab3.

Serial sections were stains with (A)monkey Ab3 (50 μg/ml) on tumor, (B) Ab1 8019 IgG1 (50 μg/ml) on tumor, (C)unrelated monkey Ab3(50 μg/ml) on tumor sections as in A or B, and (D) monkey Ab3 (50 μg/ml) on normal colon.

Chakraborty, M., Foon, K. A., Kohler, H., & Bhattacharya-Chatterjee, M. (1995). Preclinical evaluation in nonhuman primates of an anti-idiotypic antibody that mimicks the carcinoembryonic antigen. Journal of immunotherapy with emphasis on tumor immunology: official journal of the Society for Biological Therapy, 18(2), 95-103.

Figure 6 Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) pattern of ¹²⁵I-labled carcinoembryonic antigen (CEA) after immunoprecipitation.

Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) pattern of ¹²⁵I-labled carcinoembryonic antigen (CEA) after immunoprecipitation with Ab3, monkey PRO 667(lane 1), 8019(lane 2), and Ab3, monke treated with unrelated Ab2 (lane 3). The amount of antibodies esed oer lane was 10 μg.Radiolabled CEA was incubated with different Ab1 and Ab3 preparations, and the precipitated molecules was analyzed by SDS-PAGE and autoradiography. The film was exposed for 3 days.

Chakraborty, M., Foon, K. A., Kohler, H., & Bhattacharya-Chatterjee, M. (1995). Preclinical evaluation in nonhuman primates of an anti-idiotypic antibody that mimicks the carcinoembryonic antigen. Journal of immunotherapy with emphasis on tumor immunology: official journal of the Society for Biological Therapy, 18(2), 95-103.