Isolation of cancer cells by in situ microfluidic biofunctionalization protocols

Graphical abstractDisplay Omitted A microfluidic device was realized to identificate and immobilize cancer cells on biofunctionalized surfaces.The developed protocols could be used for the immobilization of cancer cells in heterogeneous samples of cells.The devices can be biofunctionalized right before the experiments when the chip is already assembled.The method could led to the realization of new devices for cancer's diagnostics in real time. The aim of this work is the development of a microfluidic immunosensor for the immobilization of cancer cells and their separation from healthy cells by using "in situ" microfluidic biofunctionalization protocols. These protocols allow to link antibodies on microfluidic device surfaces and can be used to study the interaction between cell membrane and biomolecules. Moreover they allow to perform analysis with high processing speed, small quantity of reagents and samples, short reaction times and low production costs. In this work the developed protocols were used in microfluidic devices for the isolation of cancer cells in heterogeneous blood samples by exploiting the binding of specific antibody to an adhesion protein (EpCAM), overexpressed on the tumor cell membranes. The presented biofunctionalization protocols can be performed right before running the experiment: this allows to have a flexible platform where biomolecules of interest can be linked on the device surface according to the user's needs.

[1]  Paolo A. Netti,et al.  Cell rolling and adhesion on surfaces in shear flow. A model for an antibody-based microfluidic screening system , 2012 .

[2]  Aaron M. Streets,et al.  Chip in a lab: Microfluidics for next generation life science research. , 2013, Biomicrofluidics.

[3]  Z. L. Zhang,et al.  In situ bio-functionalization and cell adhesion in microfluidic devices , 2005 .

[4]  H. Mellstedt,et al.  Epithelial cell adhesion molecule expression (CD326) in cancer: a short review. , 2012, Cancer treatment reviews.

[5]  P. Went,et al.  EpCAM expression in primary tumour tissues and metastases: an immunohistochemical analysis , 2011, Journal of Clinical Pathology.

[6]  A Manz,et al.  Protein-carbohydrate complex reveals circulating metastatic cells in a microfluidic assay. , 2013, Small.

[7]  Gerardo Perozziello,et al.  UV/Vis visible optical waveguides fabricated using organic-inorganic nanocomposite layers. , 2011, Journal of nanoscience and nanotechnology.

[8]  Suman Chakraborty,et al.  Preface to special topic: microfluidics in cancer research. , 2013, Biomicrofluidics.

[9]  Bifeng Liu,et al.  Chemiluminescence detection for a microchip capillary electrophoresis system fabricated in poly(dimethylsiloxane). , 2003, Analytical chemistry.

[10]  D. J. Harrison,et al.  Capillary electrophoresis and sample injection systems integrated on a planar glass chip , 1992 .

[11]  Rosanna La Rocca,et al.  Microfluidic biofunctionalisation protocols to form multi‐valent interactions for cell rolling and phenotype modification investigations , 2013, Electrophoresis.

[12]  N. Manaresi,et al.  A microvalve for hybrid microfluidic systems , 2010, DTIP 2010.

[13]  Gerardo Perozziello,et al.  Ca2+ Mediates the Adhesion of Breast Cancer Cells in Self-Assembled Multifunctional Microfluidic Chip Prepared with Carbohydrate Beads , 2010 .

[14]  Vijay Namasivayam,et al.  Advances in on-chip photodetection for applications in miniaturized genetic analysis systems , 2004 .

[15]  A. Al-Mehdi,et al.  Intravascular origin of metastasis from the proliferation of endothelium-attached tumor cells: a new model for metastasis , 2000, Nature Medicine.

[16]  Rosanna La Rocca,et al.  Microfluidic devices modulate tumor cell line susceptibility to NK cell recognition. , 2012, Small.

[17]  A. Jemal,et al.  Global Cancer Statistics , 2011 .

[18]  Francesco De Angelis,et al.  A microfluidic device integrating plasmonic nanodevices for Raman spectroscopy analysis on trapped single living cells , 2013 .

[19]  Frank Kohler,et al.  High‐speed chiral separations on a microchip with UV detection , 2003, Electrophoresis.

[20]  Andreas Manz,et al.  A facile in situ microfluidic method for creating multivalent surfaces: toward functional glycomics. , 2012, Lab on a chip.

[21]  Salvatore A. Pullano,et al.  A Fluidic Motherboard for Multiplexed Simultaneous and Modular Detection in Microfluidic Systems for Biological Application , 2010 .

[22]  L. Liotta,et al.  The significance of hematogenous tumor cell clumps in the metastatic process. , 1976, Cancer research.

[23]  L. Jang,et al.  Fabrication of protein chips based on 3-aminopropyltriethoxysilane as a monolayer , 2009, Biomedical microdevices.

[24]  Krist V. Gernaey,et al.  Lab on a chip automates in vitro cell culturing , 2012 .

[25]  D. J. Harrison,et al.  Planar chips technology for miniaturization and integration of separation techniques into monitoring systems. Capillary electrophoresis on a chip , 1992 .

[26]  Alison Stopeck,et al.  Circulating tumor cells, disease progression, and survival in metastatic breast cancer. , 2004, The New England journal of medicine.