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CD44-Associated Tn Antigen as a New Biomarker of Tumor Cells with Aberrant Glycosylation

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Abstract

Tn antigen is a tumor-associated antigen that appears on cancer cells as a result of aberrant O-glycosylation. The most studied form of Tn antigen is found in mucins, in particular, in mucin 1 (MUC1). Antibodies against this form of Tn antigen are used to diagnose tumors, as well as to generate T-killers with a chimeric receptor. Some carcinomas do not carry MUC1 and antibodies of a different specificity are required to detect Tn antigen on these cells. In our work, we searched for anti-Tn antibodies without preliminary assumptions about the proteins that may be carriers of the Tn antigen. For this purpose, we obtained several pairs of isogenic cell lines with the wild type and knockout of the Cosmc gene, which is essential for correct protein O-glycosylation. Using the created lines as immunogens, we generated a monoclonal antibody AKC3, which reacted with the Cosmc-deficient A549 lung adenocarcinoma cells and did not bind to the wild-type cells. Using mass spectrometry, as well as co-immunoprecipitation, it was shown that the AKC3 antibody recognized the Tn antigen in the context of CD44 protein – a protein important for tumor growth. The AKC3 antibody can be used for tumor diagnosis, and to generate T cells with a chimeric receptor for treatment of tumors that do not express mucins.

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Abbreviations

GalNAc:

N-acetyl-D-galactosamine

LRG1:

leucine rich alpha-2-glycoprotein 1; MUC1, mucin 1

MW:

molecular weight

PAGE:

polyacrylamide gel electrophoresis

PBS:

phosphate-buffered saline

SDS:

sodium dodecyl sulfate

WB:

Western blotting

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Funding

This work was financially supported by the Russian Foundation for Basic Research (project no. 18-29-07025 mk).

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Authors and Affiliations

  1. Institute of Immunology National Research Center, Federal Medical and Biological Agency of Russia, 115522, Moscow, Russia

    M. L. Shuvalova, D. V. Mazurov, A. V. Pichugin & A. V. Filatov

  2. Orekhovich Institute of Biomedical Chemistry, 119121, Moscow, Russia

    A. T. Kopylov

  3. Shemyakin and Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, 117997, Moscow, Russia

    N. V. Bovin

  4. Department of Immunology, Faculty of Biology, Lomonosov Moscow State University, 119234, Moscow, Russia

    A. V. Filatov

Authors
  1. M. L. Shuvalova
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  2. A. T. Kopylov
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  3. D. V. Mazurov
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  4. A. V. Pichugin
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  5. N. V. Bovin
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  6. A. V. Filatov
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Correspondence to A. V. Filatov.

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The authors declare no conflict of interest in financial or any other sphere. This article does not contain any studies with human participants performed by any of the authors. All applicable international, national, and/or institutional guidelines for the care and use of animals were followed.

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Shuvalova, M.L., Kopylov, A.T., Mazurov, D.V. et al. CD44-Associated Tn Antigen as a New Biomarker of Tumor Cells with Aberrant Glycosylation. Biochemistry Moscow 85, 1064–1071 (2020). https://doi.org/10.1134/S0006297920090060

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  • DOI: https://doi.org/10.1134/S0006297920090060

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