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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

Binding activity of DOTA-conjugated 11-25 mAb and 64Cu-DOTA-11-25 mAb to recombinant MSLN determined by ELISA as compared with native 11-25 mAb. Native 11-25 mAb (open triangles) and DOTA-conjugated 11-25 mAb (open squares) and 64Cu-DOTA-11-25 mAb (closed diamonds) were added to the wells on which MSLN had been immobilized. Antibody binding was detected with peroxidase-labeled anti-mouse IgG and TMB. The reaction was stopped with 2 N H2SO4 and the absorbance at 450 nm was measured.
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fig4: Binding activity of DOTA-conjugated 11-25 mAb and 64Cu-DOTA-11-25 mAb to recombinant MSLN determined by ELISA as compared with native 11-25 mAb. Native 11-25 mAb (open triangles) and DOTA-conjugated 11-25 mAb (open squares) and 64Cu-DOTA-11-25 mAb (closed diamonds) were added to the wells on which MSLN had been immobilized. Antibody binding was detected with peroxidase-labeled anti-mouse IgG and TMB. The reaction was stopped with 2 N H2SO4 and the absorbance at 450 nm was measured.

Mentions: Binding of DOTA-conjugated 11-25 mAb to the MSLN protein showed that 11-25 mAb had a similar affinity to the MSLN after conjugation with DOTA (Figure 4).


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)

Binding activity of DOTA-conjugated 11-25 mAb and 64Cu-DOTA-11-25 mAb to recombinant MSLN determined by ELISA as compared with native 11-25 mAb. Native 11-25 mAb (open triangles) and DOTA-conjugated 11-25 mAb (open squares) and 64Cu-DOTA-11-25 mAb (closed diamonds) were added to the wells on which MSLN had been immobilized. Antibody binding was detected with peroxidase-labeled anti-mouse IgG and TMB. The reaction was stopped with 2 N H2SO4 and the absorbance at 450 nm was measured.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

fig4: Binding activity of DOTA-conjugated 11-25 mAb and 64Cu-DOTA-11-25 mAb to recombinant MSLN determined by ELISA as compared with native 11-25 mAb. Native 11-25 mAb (open triangles) and DOTA-conjugated 11-25 mAb (open squares) and 64Cu-DOTA-11-25 mAb (closed diamonds) were added to the wells on which MSLN had been immobilized. Antibody binding was detected with peroxidase-labeled anti-mouse IgG and TMB. The reaction was stopped with 2 N H2SO4 and the absorbance at 450 nm was measured.
Mentions: Binding of DOTA-conjugated 11-25 mAb to the MSLN protein showed that 11-25 mAb had a similar affinity to the MSLN after conjugation with DOTA (Figure 4).

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