Biological microbeads for flow-cytometric immunoassays, enzyme titrations, and quantitative PCR

Judit Pataki, Miklós Szabó, Erika Lantos, Lóránt Székvölgyi, M. Molnár, Éva Hegedüs, Z. Bacsó, J. Kappelmayer, György Lustyik, Gábor Szabó

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

Background: Introduction of microheads into flow-cytometry has created a new scenario, making quantitative measurement of molecules dispersed in a homogeneous phase, with an extremely wide realm of already realized and potential applications possible. Development of this field has lead to specialized instrumentation and microbead arrays, dedicated to certain applications. Methods: Formaldehyde-fixed yeast and bacterial cells were conjugated with avidin and applied as microbeads, to establish a simple, convenient, flexible, and inexpensive flow-cytometric platform for various immunological and biochemical assays. Results: We have tested these "biological microbeads" for the simultaneous titration of human α-fetoprotein (AFP) and human Chorionic Gonadotropin (βhCG) hormone levels, for the titration of proteolytic and nucleolytic (restriction) enzymes, and for quantitative PCR, using biotinylated and fluorescent primers. Conclusions: The use of biological microheads for various immunological and biochemical assays has been demonstrated. The flow-cytometric methods proved to be at least as sensitive as the standard biochemical or immunological tests. For proteinase K activity measurements, a single enzyme molecule in the sample could be detected. The sensitivity, versatility, and low cost of the assays may advance flow-cytometry to become a central methodological platform in most laboratories. The biological microbeads offer virtually unlimited possibilities for fluorescent labeling (addressing), conjugation of ligand binding molecules, and they are easy to handle and perform well in a multiplex format.

Original languageEnglish
Pages (from-to)45-52
Number of pages8
JournalCytometry Part A
Volume68
Issue number1
DOIs
Publication statusPublished - Nov 2005

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Immunoenzyme Techniques
Microspheres
Polymerase Chain Reaction
Flow Cytometry
Fetal Proteins
Endopeptidase K
Immunologic Tests
Avidin
Chorionic Gonadotropin
Enzymes
Formaldehyde
Yeasts
Hormones
Ligands
Costs and Cost Analysis

Keywords

  • Biological microbeads
  • Flow cytometry
  • Immunoassay
  • Proteolytic and nucleolytic enzymes
  • Quantitative PCR

ASJC Scopus subject areas

  • Hematology
  • Cell Biology
  • Pathology and Forensic Medicine
  • Biophysics
  • Endocrinology

Cite this

Pataki, J., Szabó, M., Lantos, E., Székvölgyi, L., Molnár, M., Hegedüs, É., ... Szabó, G. (2005). Biological microbeads for flow-cytometric immunoassays, enzyme titrations, and quantitative PCR. Cytometry Part A, 68(1), 45-52. https://doi.org/10.1002/cyto.20180

Biological microbeads for flow-cytometric immunoassays, enzyme titrations, and quantitative PCR. / Pataki, Judit; Szabó, Miklós; Lantos, Erika; Székvölgyi, Lóránt; Molnár, M.; Hegedüs, Éva; Bacsó, Z.; Kappelmayer, J.; Lustyik, György; Szabó, Gábor.

In: Cytometry Part A, Vol. 68, No. 1, 11.2005, p. 45-52.

Research output: Contribution to journalArticle

Pataki, J, Szabó, M, Lantos, E, Székvölgyi, L, Molnár, M, Hegedüs, É, Bacsó, Z, Kappelmayer, J, Lustyik, G & Szabó, G 2005, 'Biological microbeads for flow-cytometric immunoassays, enzyme titrations, and quantitative PCR', Cytometry Part A, vol. 68, no. 1, pp. 45-52. https://doi.org/10.1002/cyto.20180
Pataki, Judit ; Szabó, Miklós ; Lantos, Erika ; Székvölgyi, Lóránt ; Molnár, M. ; Hegedüs, Éva ; Bacsó, Z. ; Kappelmayer, J. ; Lustyik, György ; Szabó, Gábor. / Biological microbeads for flow-cytometric immunoassays, enzyme titrations, and quantitative PCR. In: Cytometry Part A. 2005 ; Vol. 68, No. 1. pp. 45-52.
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