Quantitative Analysis of Transferrin Receptor 1 (TfR1) in Individual Breast Cancer Cells by Means of Labeled Antibodies and Elemental (ICP-MS) Detection

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Abstract
Cells are able to precisely control the amount of iron they acquire in the form of transferrin (TF)-bound iron by modulating the synthesis of the transferrin receptor 1 (TfR1). In tumor cells, elevated TfR1 seems to be related to poorer outcome for patients. Thus, the direct measurement of this biomarker in breast cancer tissues and cells might serve as a prognosis biomarker. In this work, we have used Nd-labelled antibodies to tag the TfR1 present on the cell surface of two cell models of breast cancer with different malignancy (MCF7 and MDA-MB 231). For this aim, the monoclonal antibody anti-TfR1 is first labelled with a polymeric chelator (MAXPAR®) with subsequent incorporation of several isotopic 143Nd atoms. The characterization of the labelled antibody revealed a stoichiometry of 20 Nd atoms per antibody molecule that can be used for further quantification experiments. This antibody is used for cell tagging followed by single cell analysis using inductively coupled plasma mass spectrometric (ICP-MS) detection. In this case, cell introduction is conducted using a high efficiency nebulizer and spray chamber to achieve transport efficiencies up to 55% for cells. Quantitative results revealed a number of receptors per cell significantly higher in the case of the most malignant phenotype (MDA-MB-231). Absolute and relative TfR1 concentration values are obtained in individual cells for the first time using the proposed system.
Funding Information
  • Fundación para el Fomento en Asturias de la Investigación Científica Aplicada y la Tecnología (FC-GRUPIN-IDI/2018/000242)
  • Secretaría de Estado de Investigación, Desarrollo e Innovación (CTQ2016-80069-C 2-1R, RTI2018-094605-B-I00)

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