An apta-aggregation based machine learning assay for rapid quantification of lysozyme through texture parameters.

Journal: PloS one
PMID:

Abstract

A novel assay technique that involves quantification of lysozyme (Lys) through machine learning is put forward here. This article reports the tendency of the well- documented Ellington group anti-Lys aptamer, to produce aggregates when exposed to Lys. This property of apta-aggregation has been exploited here to develop an assay that quantifies the Lys using texture and area parameters from a photograph of the elliptical aggregate mass through machine learning. Two assay sets were made for the experimental procedure: one with high Lys concentration between 25-100 mM and another with low concentration between 1-20 mM. The high concentration set had a sample volume of 10 μl while the low concentration set had a higher sample volume of 100 μl, in order to obtain the statistical texture values reliably from the aggregate mass. The platform exhibited an experimental limit of detection of 1 mM and a response time of less than 10 seconds. Further, two potential operating modes for the aptamer were hypothesized for this aggregation property and the more accurate mode among the two was ascertained through bioinformatics studies.

Authors

  • Manoharan Sanjay
    Biosensors and Bioanalysis Laboratory (LABB), Department of Biological Chemistry and IQUIBICEN-CONICET, Exact and Natural Sciences Faculty (FCEN), University of Buenos Aires (UBA), Buenos Aires, Argentina.
  • Kumar Gaurav
    Department of Biosciences and Bioengineering, Indian Institute of Technology Guwahati, Guwahati, Assam, India.
  • Maria Jesus Gonzalez-Pabon
    Biosensors and Bioanalysis Laboratory (LABB), Department of Biological Chemistry and IQUIBICEN-CONICET, Exact and Natural Sciences Faculty (FCEN), University of Buenos Aires (UBA), Buenos Aires, Argentina.
  • Julio Fuchs
    Biosensors and Bioanalysis Laboratory (LABB), Department of Biological Chemistry and IQUIBICEN-CONICET, Exact and Natural Sciences Faculty (FCEN), University of Buenos Aires (UBA), Buenos Aires, Argentina.
  • Susan R Mikkelsen
    Department of Chemistry, University of Waterloo, Waterloo, Ontario, Canada.
  • Eduardo Cortón
    Biosensors and Bioanalysis Laboratory (LABB), Department of Biological Chemistry and IQUIBICEN-CONICET, Exact and Natural Sciences Faculty (FCEN), University of Buenos Aires (UBA), Buenos Aires, Argentina.