+ Site Statistics
+ Search Articles
+ Subscribe to Site Feeds
EurekaMag Most Shared ContentMost Shared
EurekaMag PDF Full Text ContentPDF Full Text
+ PDF Full Text
Request PDF Full TextRequest PDF Full Text
+ Follow Us
Follow on FacebookFollow on Facebook
Follow on TwitterFollow on Twitter
Follow on LinkedInFollow on LinkedIn

+ Translate

A comparison of ELISA and flow microsphere-based assays for quantification of immunoglobulins

A comparison of ELISA and flow microsphere-based assays for quantification of immunoglobulins

Journal of Immunological Methods 263(1-2): 23-33

An automated microsphere-based flow cytometric assay (FlowMetrixTM system) was compared with a conventional enzymelinked immunosorbent assay (ELISA) for quantifying Ig classes in serum and stool samples. The reproducibility of the process of coupling capture antibodies to microspheres was tested. The use of independently coupled microspheres did not increase the variation of assay results relative to using the same bead set in repeated assays. However, it is necessary to ensure quality control of the coupling process since slight variations in the coupling procedures can profoundly affect the density of capture reagents coupled to the microspheres and consequently adversely affect assay precision. Although the ELISA was more sensitive and did not have the problems with instrument performance encountered with the FlowMetrixTM assay, the latter was more reproducible, had a greater dynamic range of measurement, and took considerably less preparation time than the ELISA. Greater reproducibility is especially important for measurement of fecal Ig, which is typically highly variable. Thus, in addition to its multi-analyte capability, the FlowMetrixTM assay system has definite advantages over a conventional ELISA. Mechanical problems such as microspheres settling to the bottom of wells during analysis by an automated plate reader will likely be overcome, and sensitivity improved as this technology develops.

(PDF emailed within 0-6 h: $19.90)

Accession: 010054550

Download citation: RISBibTeXText

PMID: 12009201

DOI: 10.1016/s0022-1759(02)00028-5

Related references

Quantification of human immunoglobulins Comparison of a multiplexed flow cytometric assay with ELISA. FASEB Journal 14(6): A1237, April 20, 2000

Quantification of vascular endothelial growth factor, interleukin-8, and basic fibroblast growth factor in plasma of cancer patients and healthy volunteers - comparison of ELISA and microsphere-based multiplexed immunoassay. Clinical Chemistry and Laboratory Medicine 46(9): 1256-1264, 2008

Microsphere based flow cytometric assays. Journal of Clinical Immunoassay 12(1): 36-39, 1989

Multiplexed microsphere-based flow cytometric assays. Experimental Hematology (New York) 30(11): 1227-1237, November, 2002

Quantification of myocardial blood flow and flow reserve in rats using arterial spin labeling MRI: comparison with a fluorescent microsphere technique. Nmr in Biomedicine 24(9): 1047-1053, 2012

Microsphere-based assays for genome analysis using flow cytometry. American Journal of Human Genetics 61(4 SUPPL ): A241, 1997

Superquenching as a detector for microsphere-based flow cytometric assays. Cytometry. Part A 69(5): 335-341, 2006

Multi-disease diagnostics A comparison of conventional microtiter plate-based ELISA to microsphere-based immunoassays. Abstracts of Papers American Chemical Society 224(1-2): ANYL 92, 2002

Comparison of GD2 binding capture ELISA assays for anti-GD2- antibodies using GD2-coated plates and a GD2-expressing cell-based ELISA. 2011

Comparison of GD2 binding capture ELISA assays for anti-GD2-antibodies using GD2-coated plates and a GD2-expressing cell-based ELISA. Journal of Immunological Methods 373(1-2): 181-191, 2012

Comparison of the established standard complement-dependent cytotoxicity and flow cytometric crossmatch assays with a novel ELISA-based HLA crossmatch procedure. Histology and Histopathology 21(10): 1115-1124, 2006

Cerebral blood flow quantification in the rat: a direct comparison of arterial spin labeling MRI with radioactive microsphere PET. Ejnmmi Research 2(1): 47-47, 2012

Multiplex microsphere-based flow cytometric platforms for protein analysis and their application in clinical proteomics - from assays to results. Electrophoresis 30(23): 4008-4019, 2010

Quantification of glial fibrillary acidic protein: comparison of slot-immunobinding assays with a novel sandwich ELISA. Neurotoxicology and Teratology 13(3): 275-281, 1991