μPAD Fluorescence Scattering Immunoagglutination Assay for Cancer Biomarkers from Blood and Serum

Cayla Baynes, Jeong Yeol Yoon

Research output: Contribution to journalArticlepeer-review

4 Scopus citations

Abstract

A microfluidic paper analytical device (μPAD) was created for the sensitive quantification of cancer antigens, carcinoembryonic antigen (CEA) and carbohydrate antigen 19-9 (CA 19-9), from human whole blood and serum, toward diagnosis and prognosis of colorectal cancer. Anti-CEA and anti–CA 19-9 antibodies were covalently linked to submicron, fluorescent polystyrene particles, loaded, and then dried in the center of the μPAD channel. CEA- or CA 19-9–spiked blood or serum samples were loaded to the inlet of μPAD, and subsequent immunoagglutination changed the fluorescent scatter signals upon ultraviolet (UV) excitation. The total assay time was about 1 min. Detection limits were 1 pg/mL for CEA and 0.1 U/mL for CA 19-9 from both 10% diluted blood and undiluted serum. The use of UV excitation and subsequent fluorescence scattering enabled much higher double-normalized intensities (up to 1.28–3.51, compared with 1.067 with the elastic Mie scatter detection), successful detection in the presence of blood or serum, and distinct multiplex assays with minimum cross-reaction of antibodies. The results with undiluted serum showed the larger dynamic range and smaller standard errors, which can be attributed to the presence of serum proteins, functioning as a stabilizer or a passivating protein for the particles within paper fibers.

Original languageEnglish (US)
Pages (from-to)30-43
Number of pages14
JournalSLAS Technology
Volume23
Issue number1
DOIs
StatePublished - Feb 1 2018

Keywords

  • carbohydrate antigen 19-9 (CA 19-9)
  • carcinoembryonic antigen (CEA)
  • fluorescence scatter
  • immunoagglutination
  • paper microfluidics

ASJC Scopus subject areas

  • Computer Science Applications
  • Medical Laboratory Technology

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