Capture and On-chip analysis of Melanoma Cells Using Tunable Surface Shear forces

Sci Rep. 2016 Jan 27:6:19709. doi: 10.1038/srep19709.

Abstract

With new systemic therapies becoming available for metastatic melanoma such as BRAF and PD-1 inhibitors, there is an increasing demand for methods to assist with treatment selection and response monitoring. Quantification and characterisation of circulating melanoma cells (CMCs) has been regarded as an excellent non-invasive candidate but a sensitive and efficient tool to do these is lacking. Herein we demonstrate a microfluidic approach for melanoma cell capture and subsequent on-chip evaluation of BRAF mutation status. Our approach utilizes a recently discovered alternating current electrohydrodynamic (AC-EHD)-induced surface shear forces, referred to as nanoshearing. A key feature of nanoshearing is the ability to agitate fluid to encourage contact with surface-bound antibody for the cell capture whilst removing nonspecific cells from the surface. By adjusting the AC-EHD force to match the binding affinity of antibodies against the melanoma-associated chondroitin sulphate proteoglycan (MCSP), a commonly expressed melanoma antigen, this platform achieved an average recovery of 84.7% from biological samples. Subsequent staining with anti-BRAF(V600E) specific antibody enabled on-chip evaluation of BRAF(V600E) mutation status in melanoma cells. We believe that the ability of nanoshearing-based capture to enumerate melanoma cells and subsequent on-chip characterisation has the potential as a rapid screening tool while making treatment decisions.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antibodies, Neoplasm / chemistry*
  • Antigens, Neoplasm / chemistry*
  • Cell Line, Tumor
  • Cell Separation* / instrumentation
  • Cell Separation* / methods
  • Chondroitin Sulfate Proteoglycans / chemistry*
  • Humans
  • Lab-On-A-Chip Devices*
  • Melanoma / chemistry*
  • Melanoma / genetics
  • Melanoma / metabolism
  • Melanoma / pathology
  • Mutation
  • Proto-Oncogene Proteins B-raf / genetics
  • Proto-Oncogene Proteins B-raf / metabolism

Substances

  • Antibodies, Neoplasm
  • Antigens, Neoplasm
  • Chondroitin Sulfate Proteoglycans
  • BRAF protein, human
  • Proto-Oncogene Proteins B-raf