Afuco™ Anti-Human TAG-72 ADCC Recombinant Antibody (CC49), ADCC Enhanced (CAT#: AFC-569CL)

Anti-TAG-72 ADCC Enhanced Antibody (CC49) is an ADCC enhanced antibody produced by our Afuco™ platform. CC49 is an antibody reactive to tumor-associated glycoprotein (TAG-72), has been extensively studied in targeted radionuclide therapy trials for colon cancers.


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Figure 1 Fluorescence microscopic images of LS174T cells.

Figure 1 Fluorescence microscopic images of LS174T cells.

The cells were fixed with 3.7% paraformaldehyde and incubated for 1 hour at room temperature with PBS, Cy7, murine CC49-Cy7, murine CC49 followed by murine CC49-Cy7, HuCC49ΔCH2-Cy7, and murine CC49 followed by HuCC49ΔCH2-Cy7.

Zou, P., Xu, S., Povoski, S. P., Wang, A., Johnson, M. A., Martin Jr, E. W., ... & Sun, D. (2009). Near-infrared fluorescence labeled anti-TAG-72 monoclonal antibodies for tumor imaging in colorectal cancer xenograft mice. Molecular pharmaceutics, 6(2), 428-440.

Figure 2 Whole-body imaging of subcutaneous LS174T tumor xenograft mice was accomplished by using an IVIS 100 system to monitor in vivo biodistribution of murine CC49-Cy7 and HuCC49ΔCH2-Cy7.

Figure 2 Whole-body imaging of subcutaneous LS174T tumor xenograft mice was accomplished by using an IVIS 100 system to monitor in vivo biodistribution of murine CC49-Cy7 and HuCC49ΔCH2-Cy7.

Typical in vivo fluorescence images of athymic nude mice bearing LS174T xenografts after intravenous injection of (A) 1 nmol of Cy7, excessive murine CC49 followed by 0.33 nmol of murine CC49-Cy7, 0.33 nmol of murine CC49-Cy7, or 0.38 nmol non-specific murine IgG-Cy7; (B) 1 nmol of Cy7, excessive murine CC49 followed by 0.73 nmol HuCC49ΔCH2-Cy7, 0.73 nmol HuCC49ΔCH2-Cy7, or 0.50 nmol non-specific human IgG-Cy7. The location of the tumors was indicated by the circles.

Zou, P., Xu, S., Povoski, S. P., Wang, A., Johnson, M. A., Martin Jr, E. W., ... & Sun, D. (2009). Near-infrared fluorescence labeled anti-TAG-72 monoclonal antibodies for tumor imaging in colorectal cancer xenograft mice. Molecular pharmaceutics, 6(2), 428-440.

Figure 3 The emission spectra from Cy7 and murine CC49-Cy7 solutions as well as from the tissues in the living mice were recorded with a USB4000-FL fluorescence spectrometer.

Figure 3 The emission spectra from Cy7 and murine CC49-Cy7 solutions as well as from the tissues in the living mice were recorded with a USB4000-FL fluorescence spectrometer.

Emission spectra of (A) Cy7 and murine CC49-Cy7 and (B) fluorescence signals from the bladder, liver and tumor of an athymic nude mice bearing LS174T xenograft tumor 24 hours after i.v. injection of murine CC49-Cy7.

Zou, P., Xu, S., Povoski, S. P., Wang, A., Johnson, M. A., Martin Jr, E. W., ... & Sun, D. (2009). Near-infrared fluorescence labeled anti-TAG-72 monoclonal antibodies for tumor imaging in colorectal cancer xenograft mice. Molecular pharmaceutics, 6(2), 428-440.

Figure 4 shows the relative fluorescence intensity as a function of time for the regions of interest corresponding to tumors and livers.

Figure 4 shows the relative fluorescence intensity as a function of time for the regions of interest corresponding to tumors and livers.

The relative mean fluorescence intensity in the tumor and liver regions of interest (ROIs) as a function of time after injection of (A, B) Cy7, excessive murine CC49 plus murine CC49-Cy7, or murine CC49-Cy7 and (C, D) Cy7, excessive murine CC49 plus HuCC49ΔCH2-Cy7, or HuCC49ΔCH2-Cy7.

Zou, P., Xu, S., Povoski, S. P., Wang, A., Johnson, M. A., Martin Jr, E. W., ... & Sun, D. (2009). Near-infrared fluorescence labeled anti-TAG-72 monoclonal antibodies for tumor imaging in colorectal cancer xenograft mice. Molecular pharmaceutics, 6(2), 428-440.

Figure 5 Fluorescent signals from different freshly dissected tissues were quantified by optical imaging.

Figure 5 Fluorescent signals from different freshly dissected tissues were quantified by optical imaging.

(A) Representative images of dissected organs of athymic nude mice bearing LS174T xenograft tumor sacrificed 2 hours or 96 hours after intravenous injection of 1 nmol of Cy7, excessive murine CC49 followed by 0.33 nmol of murine CC49-Cy7, 0.33 nmol of murine CC49-Cy7, or 0.38 nmol non-specific murine IgG-Cy7. The relative fluorescence intensity of each tissue was labeled near the corresponding tissue. Tissue-to-heart ratios for (B) the mice sacrificed 2 hours after injection and (C) the mice sacrificed 96 hours after injection.

Zou, P., Xu, S., Povoski, S. P., Wang, A., Johnson, M. A., Martin Jr, E. W., ... & Sun, D. (2009). Near-infrared fluorescence labeled anti-TAG-72 monoclonal antibodies for tumor imaging in colorectal cancer xenograft mice. Molecular pharmaceutics, 6(2), 428-440.

Figure 6 Fluorescent signals from different freshly dissected tissues were quantified by optical imaging.

Figure 6 Fluorescent signals from different freshly dissected tissues were quantified by optical imaging.

(A) Representative images of dissected organs of athymic nude mice bearing LS174T xenograft tumor sacrificed 2 hours or 96 hours after intravenous injection of 1 nmol of Cy7, excessive murine CC49 followed by 0.73 nmol HuCC49ΔCH2-Cy7, 0.73 nmol HuCC49ΔCH2-Cy7, or 0.50 nmol non-specific human IgG-Cy7. The relative fluorescence intensity of each tissue was labeled near the corresponding tissue. Tissue-to-heart ratios for (B) the mice sacrificed 2 hours after injection and (C) the mice sacrificed 18 hours after injection.

Zou, P., Xu, S., Povoski, S. P., Wang, A., Johnson, M. A., Martin Jr, E. W., ... & Sun, D. (2009). Near-infrared fluorescence labeled anti-TAG-72 monoclonal antibodies for tumor imaging in colorectal cancer xenograft mice. Molecular pharmaceutics, 6(2), 428-440.

Figure 7 Competitive binding assays with 3E8 and CC49.

Figure 7 Competitive binding assays with 3E8 and CC49.

Yoon, S. O., Lee, T. S., Kim, S. J., Jang, M. H., Kang, Y. J., Park, J. H., ... & Kashmiri, S. V. (2006). Construction, affinity maturation, and biological characterization of an anti-tumor-associated glycoprotein-72 humanized antibody. Journal of Biological Chemistry, 281(11), 6985-6992.

Figure 8 Immunoblot analysis of CC49-reactive protein in total cell lysates.

Figure 8 Immunoblot analysis of CC49-reactive protein in total cell lysates.

Cell lysates were prepared, resolved by SDS-PAGE, transferred to nylon membrane, then visualized with CC49 mAb. 35 µg of protein were run in each lane, except in the case of lane 8 where 50 ng purified bovine submaxillary mucin as loaded. Lane 1: LS174T xenograft cells; 2: KLE-B cells; 3: Jurkat; 4: Tf-1; 5: K562; 6: PBL; 7: M21 (human melanoma TAG-72⁻); 8: 50 ng purified bovine submaxillary mucin. The position of loading wells is indicated by an arrow. The sizes of the molecular weight standards are shown in the left. a,b Different exposures of the same filter, to show that some of darkly staining smears in the longer exposure (a) resolve to individual bands on a shorter exposure(b).

Nicolet, C. M., Siegel, D. H., Surfus, J. E., & Sondel, P. M. (1997). TAG-72-reactive antibody CC49 recognizes molecules expressed by hematopoietic cell lines. Tumor biology, 18(6), 356-366.

Figure 9 Jurket cells are targets for ADCC in the presence of chCC49.

Figure 9 Jurket cells are targets for ADCC in the presence of chCC49.

ADCC reaction were set up at three different effector: target ratios using fresh peripheral blood lymhocytes from 3 normal contorl donors (A, B and C). Both the experimental (chCC49) and contorl (ch4.18) antibodies were used at 3 µg/ml. Squares represent percent cytotoxicity in the medium alone, triangles represent percent cytotoxicity in the presence of ch4.18.

Nicolet, C. M., Siegel, D. H., Surfus, J. E., & Sondel, P. M. (1997). TAG-72-reactive antibody CC49 recognizes molecules expressed by hematopoietic cell lines. Tumor biology, 18(6), 356-366.

Figure 10 Flow cytometric analysis of cell lines using CC49 mAb.

Figure 10 Flow cytometric analysis of cell lines using CC49 mAb.

One microgram of primary antibody, murine CC49 mAb, or an IgG1 isotype control was used to stain various tissue culture lines. The fill histogram corresponds to the population stained with CC49, and the open histogram corresponds to the IgG1 isotype control. a LS174T tumor cells grown in nude mice and mechanically dispersed: the mean flourescence intensity (MFI) of the positive population (i.e. CC49-reactive) vs. the IgG1 isotype control is 39.5 vs. 9.8. b KLE endometrial carcinoma cells; MFI of the positive population vs. IgG1 isotype control is 227.4 vs. 5.1. c Jurkat T cells; MFI of the positive population vs. the IgG1 isotype control is 360.9 vs. 3.5. d Tf-1 myelomonocytic cell; MFI of the positive population vs. the IgG1 isotype control is 50.5 vs. 3.7. e K562 erythroleukemia; MFI of the positive population vs. the IgG1 isotype control is 26.5 vs. 5.0. f Unstimulated peripheral blood lymphocytes; MFI of the positive population vs. the IgG1 isotype control is 15.0 vs. 9.2 (BPL are included here as a negative contorl).

Nicolet, C. M., Siegel, D. H., Surfus, J. E., & Sondel, P. M. (1997). TAG-72-reactive antibody CC49 recognizes molecules expressed by hematopoietic cell lines. Tumor biology, 18(6), 356-366.


Specifications

  • Host Species
  • Humanized
  • Derivation
  • Humanized
  • Type
  • ADCC enhanced antibody
  • Species Reactivity
  • Human
  • Related Disease
  • Cancer

Product Property

  • Purity
  • >97%, by SDS-PAGE under reducing conditions
  • Storage
  • Store at –20 or -70°C upon receipt. Divide antibody into aliquots prior usage and avoid multiple freeze-thaw cycles.

Target

  • Alternative Names
  • TAG-72; Tumor-associated glycoprotein 72

Product Notes

This is a product of Creative Biolabs' Hi-Affi™ recombinant antibody portfolio, which has several benefits including:

• Increased sensitivity
• Confirmed specificity
• High repeatability
• Excellent batch-to-batch consistency
• Sustainable supply
• Animal-free production

See more details about Hi-Affi™ recombinant antibody benefits.

Downloads

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See other products for "TAG72"

Mouse Antibody

CAT Product Name Application Type
TAB-723 Mouse Anti-TAG72 Recombinant Antibody (clone Anatumomab); Fab Fragment FuncS, IF, Neut, ELISA, FC, IP, WB Mouse Fab (IgG1, κ)
TAB-H60 Anti-Human TAG-72 Recombinant Antibody (Satumomab) ELISA, FC, IP, FuncS, IF, Neut, ICC IgG1 - kappa
TAB-249 Anti-Human TAG-72 Recombinant Antibody (Minretumomab) WB, FuncS, IF, Neut, ELISA, FC, IP IgG1
TAB-002 Anti-Human TAG72 Recombinant Antibody (Satumomab Pendetide) IP, IF, FuncS, FC, Neut, ELISA, WB IgG1
PABX-025-S (P) Recombinant Human Anti-TAG-72 Antibody scFv Fragment (B72.3) WB, Neut, FuncS scFv

Immunotoxin

CAT Product Name Application Type
AGTO-L070E anti-TAG72 immunotoxin CC49 (scFv)-PE Cytotoxicity assay, Functional assay

Fc Glycosylation

CAT Product Name Application Type
Gly-016LC Recombinant Anti-Human TAG-72 Antibody (Fc glycosylation) ELISA Chimeric antibody (mouse/human)

ADCC Enhanced Antibody

CAT Product Name Application Type
AFC-TAB-249 Afuco™ Anti-TAG72 ADCC Recombinant Antibody (Minretumomab), ADCC Enhanced FuncS, IF, Neut, ELISA, FC ADCC enhanced antibody
AFC-TAB-H60 Afuco™ Anti-TAG72 ADCC Recombinant Antibody (Satumomab), ADCC Enhanced ELISA, FC, IP, FuncS, IF, Neut ADCC enhanced antibody

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