Surface-decorated S. cerevisiae for flow cytometric array immunoassay

Surface-decorated S. cerevisiae for flow cytometric array immunoassay Our laboratory had developed a cell-based bio-bead for protein quantification. However, the selection of antibody in the above immunoassay is limited. This study describes a surface-decorated Saccharomyces cerevisiae for flow cytometric array immunoassay. S. cerevisiae was labeled with fluorescein isothiocyanate (FITC) and oxidized by sodium periodate, in which the saccharide group on the cytoderm outer layer was converted to an aldehyde group. In succession, adipic dihydrazide was bio-conjugated to the aldehyde group and glutaraldehyde bound to the hydrazide group. Phycoerythrin (PE)-labeled goat anti-mouse polyclonal antibody was used to assess the conjugation of mouse anti-human monoclonal antibody to surface-decorated S. cerevisiae. Cytokeratin 19 fragment (Cyfra21-1) and neuron-specific enolase (NSE) antigens were also employed to evaluate the flow cytometric array immunoassay based on surface-decorated S. cerevisiae. Flow cytometry demonstrated that FITC-barcoded S. cerevisiae as two legible populations. PE-labeled polyclonal antibody validated the coating of surface-decorated S. cerevisiae with the monoclonal antibody. The flow cytometric array immunoassays for Cyfra21-1 and NSE documented that the limit of detection (LOD) was at least 0.4 ng/mL. Precision and accuracy assessments appeared that the relative standard deviation (R.S.D.) was <15%, and the relative error (R.E.) ranged from 0.9 to 1.1. The correlation coefficient between this immunoassay and electrochemiluminescence immunoassay was 0.9622 for serum Cyfra21-1 and 0.9918 for serum NSE. In conclusion, the surface-decorated S. cerevisiae may be of use in flow cytometric array immunoassay. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Analytical and Bioanalytical Chemistry Springer Journals

Surface-decorated S. cerevisiae for flow cytometric array immunoassay

Loading next page...
 
/lp/springer_journal/surface-decorated-s-cerevisiae-for-flow-cytometric-array-immunoassay-rQqnq00jyE
Publisher
Springer Berlin Heidelberg
Copyright
Copyright © 2017 by Springer-Verlag GmbH Germany
Subject
Chemistry; Analytical Chemistry; Biochemistry, general; Laboratory Medicine; Characterization and Evaluation of Materials; Food Science; Monitoring/Environmental Analysis
ISSN
1618-2642
eISSN
1618-2650
D.O.I.
10.1007/s00216-017-0470-z
Publisher site
See Article on Publisher Site

Abstract

Our laboratory had developed a cell-based bio-bead for protein quantification. However, the selection of antibody in the above immunoassay is limited. This study describes a surface-decorated Saccharomyces cerevisiae for flow cytometric array immunoassay. S. cerevisiae was labeled with fluorescein isothiocyanate (FITC) and oxidized by sodium periodate, in which the saccharide group on the cytoderm outer layer was converted to an aldehyde group. In succession, adipic dihydrazide was bio-conjugated to the aldehyde group and glutaraldehyde bound to the hydrazide group. Phycoerythrin (PE)-labeled goat anti-mouse polyclonal antibody was used to assess the conjugation of mouse anti-human monoclonal antibody to surface-decorated S. cerevisiae. Cytokeratin 19 fragment (Cyfra21-1) and neuron-specific enolase (NSE) antigens were also employed to evaluate the flow cytometric array immunoassay based on surface-decorated S. cerevisiae. Flow cytometry demonstrated that FITC-barcoded S. cerevisiae as two legible populations. PE-labeled polyclonal antibody validated the coating of surface-decorated S. cerevisiae with the monoclonal antibody. The flow cytometric array immunoassays for Cyfra21-1 and NSE documented that the limit of detection (LOD) was at least 0.4 ng/mL. Precision and accuracy assessments appeared that the relative standard deviation (R.S.D.) was <15%, and the relative error (R.E.) ranged from 0.9 to 1.1. The correlation coefficient between this immunoassay and electrochemiluminescence immunoassay was 0.9622 for serum Cyfra21-1 and 0.9918 for serum NSE. In conclusion, the surface-decorated S. cerevisiae may be of use in flow cytometric array immunoassay.

Journal

Analytical and Bioanalytical ChemistrySpringer Journals

Published: Jul 4, 2017

References

You’re reading a free preview. Subscribe to read the entire article.


DeepDyve is your
personal research library

It’s your single place to instantly
discover and read the research
that matters to you.

Enjoy affordable access to
over 18 million articles from more than
15,000 peer-reviewed journals.

All for just $49/month

Explore the DeepDyve Library

Search

Query the DeepDyve database, plus search all of PubMed and Google Scholar seamlessly

Organize

Save any article or search result from DeepDyve, PubMed, and Google Scholar... all in one place.

Access

Get unlimited, online access to over 18 million full-text articles from more than 15,000 scientific journals.

Your journals are on DeepDyve

Read from thousands of the leading scholarly journals from SpringerNature, Elsevier, Wiley-Blackwell, Oxford University Press and more.

All the latest content is available, no embargo periods.

See the journals in your area

DeepDyve

Freelancer

DeepDyve

Pro

Price

FREE

$49/month
$360/year

Save searches from
Google Scholar,
PubMed

Create lists to
organize your research

Export lists, citations

Read DeepDyve articles

Abstract access only

Unlimited access to over
18 million full-text articles

Print

20 pages / month

PDF Discount

20% off