Limits...
A novel PET imaging using ⁶⁴Cu-labeled monoclonal antibody against mesothelin commonly expressed on cancer cells.

Kobayashi K, Sasaki T, Takenaka F, Yakushiji H, Fujii Y, Kishi Y, Kita S, Shen L, Kumon H, Matsuura E - J Immunol Res (2015)

Bottom Line: Mesothelin (MSLN) is a 40-kDa cell differentiation-associated glycoprotein appearing with carcinogenesis and is highly expressed in many human cancers, including the majority of pancreatic adenocarcinomas, ovarian cancers, and mesotheliomas, while its expression in normal tissue is limited to mesothelial cells lining the pleura, pericardium, and peritoneum.Clone 11-25 is a murine hybridoma secreting monoclonal antibody (mAb) against human MSLN.In in vitro and ex vivo immunochemical studies, we demonstrated specificity of 11-25 mAb to membranous MSLN expressed on several pancreatic cancer cells.

View Article: PubMed Central - PubMed

Affiliation: Department of Cell Chemistry, Okayama University Graduate School of Medicine, Dentistry, and Pharmaceutical Sciences, Okayama 700-8558, Japan ; Department of Urology, Okayama University Graduate School of Medicine, Dentistry, and Pharmaceutical Sciences, Okayama 700-8558, Japan.

ABSTRACT
Mesothelin (MSLN) is a 40-kDa cell differentiation-associated glycoprotein appearing with carcinogenesis and is highly expressed in many human cancers, including the majority of pancreatic adenocarcinomas, ovarian cancers, and mesotheliomas, while its expression in normal tissue is limited to mesothelial cells lining the pleura, pericardium, and peritoneum. Clone 11-25 is a murine hybridoma secreting monoclonal antibody (mAb) against human MSLN. In this study, we applied the 11-25 mAb to in vivo imaging to detect MSLN-expressing tumors. In in vitro and ex vivo immunochemical studies, we demonstrated specificity of 11-25 mAb to membranous MSLN expressed on several pancreatic cancer cells. We showed the accumulation of Alexa Fluor 750-labeled 11-25 mAb in MSLN-expressing tumor xenografts in athymic nude mice. Then, 11-25 mAb was labeled with (64)Cu via a chelating agent DOTA and was used in both in vitro cell binding assay and in vivo positron emission tomography (PET) imaging in the tumor-bearing mice. We confirmed that (64)Cu-labeled 11-25 mAb highly accumulated in MSLN-expressing tumors as compared to MSLN-negative ones. The (64)Cu-labeled 11-25 mAb is potentially useful as a PET probe capable of being used for wide range of tumors, rather than (18)F-FDG that occasionally provides nonspecific accumulation into the inflammatory lesions.

No MeSH data available.


Related in: MedlinePlus

Cell binding assay with 64Cu-DOTA-11-25 mAb. BxPC-3 cells were transferred to 24-well plates at 5 × 104 cells/well/mL and cultured for 4 days. Various concentrations of 64Cu-DOTA-11-25 mAb were incubated with the cell monolayers for 2 hours on ice in complete growth media (RPMI-1640 medium containing 10% fetal bovine serum). The cells were washed and lysed and their radioactivity was counted in a γ-counter. Inner graph showed the Scatchard plot of the specific binding versus the concentration of 64Cu-DOTA-11-25 mAb.
© Copyright Policy - open-access
Related In: Results  -  Collection


getmorefigures.php?uid=PMC4390102&req=5

fig6: Cell binding assay with 64Cu-DOTA-11-25 mAb. BxPC-3 cells were transferred to 24-well plates at 5 × 104 cells/well/mL and cultured for 4 days. Various concentrations of 64Cu-DOTA-11-25 mAb were incubated with the cell monolayers for 2 hours on ice in complete growth media (RPMI-1640 medium containing 10% fetal bovine serum). The cells were washed and lysed and their radioactivity was counted in a γ-counter. Inner graph showed the Scatchard plot of the specific binding versus the concentration of 64Cu-DOTA-11-25 mAb.

Mentions: For in vitro experiment, 64Cu-DOTA-11-25 mAb was prepared and a binding study of 64Cu-DOTA-11-25 mAb with alive BxPC-3 cells was performed at 4°C. BxPC-3 cells exhibited saturable 64Cu-DOTA-11-25 mAb binding (Figure 6). The KD of 64Cu-DOTA-11-25 mAb binding for the MSLN on BxPC-3 cells was 353 ± 63 nM.


A novel PET imaging using ⁶⁴Cu-labeled monoclonal antibody against mesothelin commonly expressed on cancer cells.

Kobayashi K, Sasaki T, Takenaka F, Yakushiji H, Fujii Y, Kishi Y, Kita S, Shen L, Kumon H, Matsuura E - J Immunol Res (2015)

Cell binding assay with 64Cu-DOTA-11-25 mAb. BxPC-3 cells were transferred to 24-well plates at 5 × 104 cells/well/mL and cultured for 4 days. Various concentrations of 64Cu-DOTA-11-25 mAb were incubated with the cell monolayers for 2 hours on ice in complete growth media (RPMI-1640 medium containing 10% fetal bovine serum). The cells were washed and lysed and their radioactivity was counted in a γ-counter. Inner graph showed the Scatchard plot of the specific binding versus the concentration of 64Cu-DOTA-11-25 mAb.
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4390102&req=5

fig6: Cell binding assay with 64Cu-DOTA-11-25 mAb. BxPC-3 cells were transferred to 24-well plates at 5 × 104 cells/well/mL and cultured for 4 days. Various concentrations of 64Cu-DOTA-11-25 mAb were incubated with the cell monolayers for 2 hours on ice in complete growth media (RPMI-1640 medium containing 10% fetal bovine serum). The cells were washed and lysed and their radioactivity was counted in a γ-counter. Inner graph showed the Scatchard plot of the specific binding versus the concentration of 64Cu-DOTA-11-25 mAb.
Mentions: For in vitro experiment, 64Cu-DOTA-11-25 mAb was prepared and a binding study of 64Cu-DOTA-11-25 mAb with alive BxPC-3 cells was performed at 4°C. BxPC-3 cells exhibited saturable 64Cu-DOTA-11-25 mAb binding (Figure 6). The KD of 64Cu-DOTA-11-25 mAb binding for the MSLN on BxPC-3 cells was 353 ± 63 nM.

Bottom Line: Mesothelin (MSLN) is a 40-kDa cell differentiation-associated glycoprotein appearing with carcinogenesis and is highly expressed in many human cancers, including the majority of pancreatic adenocarcinomas, ovarian cancers, and mesotheliomas, while its expression in normal tissue is limited to mesothelial cells lining the pleura, pericardium, and peritoneum.Clone 11-25 is a murine hybridoma secreting monoclonal antibody (mAb) against human MSLN.In in vitro and ex vivo immunochemical studies, we demonstrated specificity of 11-25 mAb to membranous MSLN expressed on several pancreatic cancer cells.

View Article: PubMed Central - PubMed

Affiliation: Department of Cell Chemistry, Okayama University Graduate School of Medicine, Dentistry, and Pharmaceutical Sciences, Okayama 700-8558, Japan ; Department of Urology, Okayama University Graduate School of Medicine, Dentistry, and Pharmaceutical Sciences, Okayama 700-8558, Japan.

ABSTRACT
Mesothelin (MSLN) is a 40-kDa cell differentiation-associated glycoprotein appearing with carcinogenesis and is highly expressed in many human cancers, including the majority of pancreatic adenocarcinomas, ovarian cancers, and mesotheliomas, while its expression in normal tissue is limited to mesothelial cells lining the pleura, pericardium, and peritoneum. Clone 11-25 is a murine hybridoma secreting monoclonal antibody (mAb) against human MSLN. In this study, we applied the 11-25 mAb to in vivo imaging to detect MSLN-expressing tumors. In in vitro and ex vivo immunochemical studies, we demonstrated specificity of 11-25 mAb to membranous MSLN expressed on several pancreatic cancer cells. We showed the accumulation of Alexa Fluor 750-labeled 11-25 mAb in MSLN-expressing tumor xenografts in athymic nude mice. Then, 11-25 mAb was labeled with (64)Cu via a chelating agent DOTA and was used in both in vitro cell binding assay and in vivo positron emission tomography (PET) imaging in the tumor-bearing mice. We confirmed that (64)Cu-labeled 11-25 mAb highly accumulated in MSLN-expressing tumors as compared to MSLN-negative ones. The (64)Cu-labeled 11-25 mAb is potentially useful as a PET probe capable of being used for wide range of tumors, rather than (18)F-FDG that occasionally provides nonspecific accumulation into the inflammatory lesions.

No MeSH data available.


Related in: MedlinePlus