Open AccessMore sensitive immune complex transfer enzyme immunoassay for antibody IgG to p17 of HIV‐1 with shorter incubation time for immunoreactions and larger volumes of serum samples
Author(s) -
Ishikawa Setsuko,
Hashida Seiichi,
Hashinaka Kazuya,
Kojima Masaaki,
Saito Atsushi,
Takamizawa Akihisa,
Shinagawa Hideo,
Oka Shinichi,
Shimada Kaoru,
Ishikawa Eiji
Publication year - 1997
Publication title -
journal of clinical laboratory analysis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.536
H-Index - 50
eISSN - 1098-2825
pISSN - 0887-8013
DOI - 10.1002/(sici)1098-2825(1997)11:5<244::aid-jcla2>3.0.co;2-7
Subject(s) - incubation , immunoassay , antibody , bovine serum albumin , dinitrophenyl , conjugate , chemistry , incubation period , chromatography , immunoglobulin g , immune complex , microbiology and biotechnology , biology , immunology , biochemistry , mathematical analysis , mathematics
In the previous immune complex transfer enzyme immunoassay for antibody IgG to p17 of HIV‐1, the immune complex comprising 2,4‐dinitrophenyl‐bovine serum albumin‐recombinant p17 conjugate, anti‐p17 IgG, and recombinant p17‐β‐D‐galactosidase conjugate was trapped onto polystyrene beads coated with (anti‐2,4‐dinitrophenyl group) IgG by overnight incubation and was transferred to polystyrene beads coated with (antihuman IgG γ‐chain) IgG by 3 hr incubation in the presence of excess of ϵ N ‐2,4‐dinitrophenyl‐ L ‐lysine. These processes were made efficient by incubation with shaking and by using solid phases with larger surface areas. In addition, the volume of serum samples used was increased from 10 μl to 100 μl. As a result, the sensitivity was improved 20–30‐fold and was ˜100,000‐fold higher than that of Western blotting for p17 band, even when both trapping and transferring of the immune complex were performed for only 30 min. Furthermore, testing many samples became easily possible with higher sensitivity using microplates and a fluororeader. J. Clin. Lab. Anal. 11:244–250, 1997. © 1997 Wiley‐Liss, Inc. No abstract.