Highly sensitive time-resolved fluoroimmunoassay of human immunoglobulin E by using a new europium fluorescent chelate as a label

Analytical Biochemistry
J YuanKazuko Matsumoto

Abstract

A new europium fluorescent chelate, 4,4'-bis(1",1",1",2",2",3",3"-heptafluoro-4",6"-hexanedione-6"-yl) - chlorosulfo-o-terphenyl (BHHCT)-Eu3+, was used as a label for highly sensitive time-resolved fluoroimmunoassay of human IgE. Two assay formats were employed in the analysis. In the first format, an immunoconjugate of rabbit anti-human IgE antibody-human IgE-biotinylated goat anti-human IgE antibody-BHHCT-Eu(3+)-labeled SA (or BHHCT-Eu(3+)-labeled BSA-SA; BSA, bovine serum albumin; SA, streptavidin) was used for measurement. The method gives the detection limits of 3.6 x 10(-2) IU/ml (labeled SA) and 1.1 x 10(-2) IU/ml (labeled SA-BSA). In the second format, an immunoconjugate of goat anti-human IgE antibody-human IgE-rabbit anti-human IgE antibody-biotinylated goat anti-rabbit IgG antibody-BHHCT-Eu(3+)-labeled SA (or BHHCT-Eu(3+)-labeled BSA-SA) was used for measurement. The detection limits of these methods are 3.0 x 10(-3) IU/ml (labeled SA) and 1.5 x 10(-3) IU/ml (labeled BSA-SA). The above detection limits are one to two orders of magnitude lower than those of the conventional radioimmunoassay and enzyme immunoassay. The CV of the present method is less than 7%, and the recovery is in the range of 85-105% for serum samples.

References

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Citations

Mar 1, 2003·Analytical Sciences : the International Journal of the Japan Society for Analytical Chemistry·Michimasa KobayashiYasuhiko Tomino
Apr 2, 2004·Analytical Sciences : the International Journal of the Japan Society for Analytical Chemistry·Xiaodan HaiKazuko Matsumoto
Aug 31, 2002·Analytical Sciences : the International Journal of the Japan Society for Analytical Chemistry·Keisuke MajimaKazuko Matsumoto
Nov 16, 2001·Analytical Sciences : the International Journal of the Japan Society for Analytical Chemistry·G WangH Kimura
Apr 5, 2011·Cytometry. Part a : the Journal of the International Society for Analytical Cytology·Yiqing LuYujing Huo
Mar 20, 2010·Physical Chemistry Chemical Physics : PCCP·Fumin XueYuan Wang
Sep 1, 2005·Luminescence : the Journal of Biological and Chemical Luminescence·Zhen WangKazuko Matsumoto
Sep 17, 2005·Journal of Fluorescence·Jingli Yuan, Guilan Wang
Nov 15, 2005·Chemical Society Reviews·Jean-Claude G Bünzli, Claude Piguet
Mar 18, 2011·Dalton Transactions : an International Journal of Inorganic Chemistry·Ping LianChunhui Huang
Jan 23, 2014·Methods and Applications in Fluorescence·Johanna Vuojola, Tero Soukka
Apr 22, 2010·Langmuir : the ACS Journal of Surfaces and Colloids·Wei DengEwa M Goldys
Sep 9, 2000·Forensic Science International : Synergy·H KimuraK Matsumoto
May 9, 2006·Inorganic Chemistry·Takuya NishiokaKazuko Matsumoto

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