Jump to main content
Jump to site search

Issue 44, 2016
Previous Article Next Article

Differential quantification of SCCA1 and SCCA2 cancer antigens using a hydrogel biochip

Author affiliations

Abstract

Methods employing hydrogel-based microarrays (biochips) allow the simultaneous monitoring of protein interactions with different antibodies immobilized in gel elements. The method was applied for the simultaneous differential quantification of two highly homologous antigens of squamous cell carcinomas (SCCs) SCCA1 and SCCA2 in a single analysis. Two panels of monoclonal antibodies against recombinant SCCA1 and SCCA2 were generated, and two antibodies, C5 (anti-SCCA1) and A11 (anti-SCCA2), were selected for further evaluation based on their ability to specifically interact with their cognate antigens. Using a sandwich analysis, these antibodies were further tested in combination with anti-SCCA antibodies (H31 and SCC107) recognizing both of the SCCA antigens, thus allowing a quantitative independent measurement of both antigens. The intra- and inter-assay coefficients of variation for all resultant tests did not exceed 10% for the range of SCCA concentrations tested and were independent of whether SCCA1 and SCCA2 concentrations were determined simultaneously. The lower limit of detection (LOD) was estimated as 0.006 ng ml−1 for SCCA1 and 0.011 ng ml−1 for SCCA2 using the SCC107-Cy5 developing antibody and 0.014 ng ml−1 and 0.01 ng ml−1 concentrations, respectively, of the H31-Cy5 developing antibody. This assay provides a simple and accurate procedure for the differential quantitation of SCCA1 and SCCA2 using a single analysis of human serum on a biochip.

Graphical abstract: Differential quantification of SCCA1 and SCCA2 cancer antigens using a hydrogel biochip

Back to tab navigation
Please wait while Download options loads

Publication details

The article was received on 05 Aug 2016, accepted on 03 Oct 2016 and first published on 07 Oct 2016


Article type: Paper
DOI: 10.1039/C6AY02216B
Citation: Anal. Methods, 2016,8, 7920-7928
  • Open access: Creative Commons BY license
  •   Request permissions

    Differential quantification of SCCA1 and SCCA2 cancer antigens using a hydrogel biochip

    A. A. Tikhonov, M. V. Tsybulskaya, V. I. Butvilovskaya, E. N. Savvateeva, P. V. Belousov, D. V. Kuprash, O. N. Solopova, M. A. Chernichenko, M. M. Filushin and A. Yu. Rubina, Anal. Methods, 2016, 8, 7920
    DOI: 10.1039/C6AY02216B

    This article is licensed under a Creative Commons Attribution 3.0 Unported Licence. Material from this article can be used in other publications provided that the correct acknowledgement is given with the reproduced material.

    Reproduced material should be attributed as follows:

    • For reproduction of material from NJC:
      [Original citation] - Published by The Royal Society of Chemistry (RSC) on behalf of the Centre National de la Recherche Scientifique (CNRS) and the RSC.
    • For reproduction of material from PCCP:
      [Original citation] - Published by the PCCP Owner Societies.
    • For reproduction of material from PPS:
      [Original citation] - Published by The Royal Society of Chemistry (RSC) on behalf of the European Society for Photobiology, the European Photochemistry Association, and RSC.
    • For reproduction of material from all other RSC journals:
      [Original citation] - Published by The Royal Society of Chemistry.

    Information about reproducing material from RSC articles with different licences is available on our Permission Requests page.

Search articles by author