Monoclonal antibody MX35 detects the membrane transporter NaPi2b (SLC34A2) in human carcinomas

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Monoclonal antibody MX35 detects the membrane transporter NaPi2b (SLC34A2) in human carcinomas

Yin, Beatrice W T (author): Ludwig Institute for Cancer Research, New York Branch at Memorial Sloan-Kettering Cancer Center, New York, NY, USA

Kiyamova, Ramziya (author): The Laboratory of Cell Growth Regulation, Department of Cell Signaling, Institute of Molecular Biology and Genetics, Kyiv, Ukraine

Chua, Ramon (author): Ludwig Institute for Cancer Research, New York Branch at Memorial Sloan-Kettering Cancer Center, New York, NY, USA

Caballero, Otavia L (author): Ludwig Institute for Cancer Research, New York Branch at Memorial Sloan-Kettering Cancer Center, New York, NY, USA

Gout, Ivan (author): Department of Biochemistry and Molecular Biology, University College London, London, United Kingdom

Gryshkova, Vitalina (author): The Laboratory of Cell Growth Regulation, Department of Cell Signaling, Institute of Molecular Biology and Genetics, Kyiv, Ukraine

Bhaskaran, Nimesh (author): Uppsala universitet,Ludwiginstitutet för cancerforskning

Souchelnytskyi, Serhiy (author): Uppsala universitet,Ludwiginstitutet för cancerforskning

Hellman, Ulf (author): Uppsala universitet,Ludwiginstitutet för cancerforskning

Filonenko, Valeriy (author): The Laboratory of Cell Growth Regulation, Department of Cell Signaling, Institute of Molecular Biology and Genetics, Kyiv, Ukraine

Jungbluth, Achim A. (author): Ludwig Institute for Cancer Research, New York Branch at Memorial Sloan-Kettering Cancer Center, New York, NY, USA

Odunsi, Kunle (author): Department of Gynecologic Oncology, Roswell Park Cancer Institute, Buffalo, NY, USA

Lloyd, Kenneth O (author): Immunology Program, Memorial Sloan-Kettering Cancer Center, New York, NY, USA

Old, Lloyd J (author): Ludwig Institute for Cancer Research, New York Branch at Memorial Sloan-Kettering Cancer Center, New York, NY, USA

Ritter, Gerd (author): Ludwig Institute for Cancer Research, New York Branch at Memorial Sloan-Kettering Cancer Center, New York, NY, USA

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2008
2008
English.
In: Cancer Immunity. - 1424-9634. ; 8, s. 3-

Related links:: http://www.cancerimm...; show more...; https://urn.kb.se/re...; show less...

Journal article (peer-reviewed)

Abstract Subject headings

Mouse monoclonal antibody MX35 was developed against ovarian cancer. The antibody showed homogeneous reactivity with approximately 90% of human ovarian epithelial cancers and with a limited number of normal tissues by immunohistochemistry. Although mAb MX35 has been used in a number of clinical trials in ovarian cancer, it has been difficult to define the molecular identity of MX35. We report here that mAb MX35 recognizes the sodium-dependent phosphate transport protein 2b (NaPi2b) in human cancer cells. This conclusion is based on several lines of experimental evidence, including 1) the identification of SLC34A2, the gene coding for NaPi2b, by immunoscreening an ovarian cancer cell line cDNA expression library with mAb MX35; 2) mass spectrometry sequencing of peptides obtained by fragmentation from mAb MX35 affinity-purified antigen, which show complete sequence homology to amino acid sequences in NaPi2b; 3) selective down-regulation of SLC34A2 gene expression by RNA interference and the resulting loss of mAb MX35 binding to MX35-expressing human cancer cells; and 4) the demonstration of specific mAb MX35 reactivity with recombinant fusion proteins and with synthetic peptides of the putative largest extracellular loop of NaPi2b. We further show that NaPi2b in cancer cells is expressed on the cell surface as a heavily N-glycosylated protein, with evidence of additional post-translational modifications such as palmitoylation and the formation of disulfide bridges in the major extracellular loop. Membrane transporter molecules, such as NaPi2b, represent a new family of potential cell surface targets for the immunotherapy of cancer with monoclonal antibodies.

Keyword

ovarian cancer
monoclonal antibody
MX35
protein sequence analysis
NaPi2b
tumor antigen
cancer immunotherapy
MEDICINE
MEDICIN

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By the author/editor: Yin, Beatrice W ...; Kiyamova, Ramziy ...; Chua, Ramon; Caballero, Otavi ...; Gout, Ivan; Gryshkova, Vital ...; show more...; Bhaskaran, Nimes ...; Souchelnytskyi, ...; Hellman, Ulf; Filonenko, Valer ...; Jungbluth, Achim ...; Odunsi, Kunle; Lloyd, Kenneth O; Old, Lloyd J; Ritter, Gerd; show less...

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By the university: Uppsala University

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