Journal article
Analysis of antigen‐specific antibodies and their isotypes in experimental malaria
Cytometry. Part A, v 71A(4)
Apr 2007
PMID: 17252581
Featured in Collection : UN Sustainable Development Goals @ Drexel
Abstract
Background
Measuring antibody production in response to antigen exposure or vaccination is key to disease prevention and treatment. Our understanding of the mechanisms involved in the antibody response is limited by a lack of sensitive analysis methods. We address this limitation using multiplexed microsphere arrays for the semi ‐quantitative analysis of antibody production in response to malaria infection.
Methods
We used microspheres as solid supports on which to capture and analyze circulating antibodies. Antigen immobilized on beads captured antigen‐specific antibodies for semi‐ quantitative analysis using fluorescent secondary antibodies. Anti‐immunoglobulin antibodies on beads captured specific antibody isotypes for affinity estimation using fluorescent antigen.
Results
Antigen‐mediated capture of plasma antibodies enables determination of antigen‐specific antibody “titer,” a semi‐quantitative parameter describing a convolution of antibody abundance and avidity, as well as parameters describing numbers of antibodies bound/bead at saturation and the plasma concentration‐dependent approach to saturation. Results were identical in single‐plex and multiplex assays, and in qualitative agreement with similar parameters derived from ELISA‐based assays. Isotype‐specific antibody‐mediated capture of plasma antibodies allowed the estimation of the affinity of antibody for antigen.
Conclusion
Analysis of antibody responses using microspheres and flow cytometry offer significant advantages in speed, sample size, and quantification over standard ELISA‐based titer methods. © 2007 International Society for Analytical Cytology.
Metrics
Details
- Title
- Analysis of antigen‐specific antibodies and their isotypes in experimental malaria
- Creators
- Henri C van der HeydeJames M BurnsWilliam P WeidanzJohn HornIrene GramagliaJohn P Nolan
- Publication Details
- Cytometry. Part A, v 71A(4)
- Publisher
- Wiley Subscription Services, Inc., A Wiley Company; Hoboken
- Number of pages
- 9
- Grant note
- NIH (AI40667; AI12710; AI49585; EB003824)
- Resource Type
- Journal article
- Language
- English
- Academic Unit
- Microbiology and Immunology
- Web of Science ID
- WOS:000245343200005
- Scopus ID
- 2-s2.0-34147219598
- Other Identifier
- 991014877871804721
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- Collaboration types
- Domestic collaboration
- Web of Science research areas
- Biochemical Research Methods
- Cell Biology