Lectin-dependent inhibition of antigen-antibody reaction: application for measuring α2,6-sialylated glycoforms of transferrin.
We developed a high-throughput Enzyme-linked immunosorbent assay (ELISA) for measuring α2,6-sialylated transferrin (Tf), based on inhibition of anti-Tf antibody binding to α2,6-sialylated Tf by a lectin, Sambucus sieboldiana Agglutinin (SSA). The inhibition was not observed with other glycoforms, such as periodate-treated, sialidase-treated and sialidase/galactosidase-treated Tf, suggesting that the assay was glycoform specific. This finding was applied to an automated latex-agglutination immunoassay, using SSA lectin as an inhibitor (SSA-ALI). The concentration of α2,6-sialylated Tf measured by SSA-ALI in human cerebrospinal fluid was correlated with that of ELISA (r2 = 0.8554), previously developed for measuring α2,6-sialylated Tf.
Department of Biochemistry, Fukushima Medical University School of Medicine, 1 Hikarigaoka, Fukushima-shi, Fukushima 960-1295, Japan.
Kyoka
K
Hoshi
Yoshinobu
Y
Kariya
Kiyomitsu
K
Nara
Hiromi
H
Ito
Kana
K
Matsumoto
Masamichi
M
Nagae
Yoshiki
Y
Yamaguchi
Madoka
M
Nakajima
Masakazu
M
Miyajima
Hajime
H
Arai
Atsushi
A
Kuno
Hisashi
H
Narimatsu
Keiro
K
Shirotani
Yasuhiro
Y
Hashimoto
D000818:Animals; D000937:Antigen-Antibody Reactions / Q000276:immunology*; D001667:Binding, Competitive; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D006023:Glycoproteins / Q000134:cerebrospinal fluid*; D006031:Glycosylation; D006041:Goats; D057166:High-Throughput Screening Assays; D006801:Humans; D009439:Neuraminidase / Q000737:chemistry; D037121:Plant Lectins / Q000737:chemistry* / Q000276:immunology; D011485:Protein Binding; D011817:Rabbits; D054788:Ribosome Inactivating Proteins / Q000737:chemistry* / Q000276:immunology; D014168:Transferrin / Q000134:cerebrospinal fluid*
D006023:Glycoproteins; D037121:Plant Lectins; C050600:Sambucus nigra lectins; D014168:Transferrin; D009439:Neuraminidase; D054788:Ribosome Inactivating Proteins
Sambucus sieboldiana Agglutinin lectin; automated latex-agglutination immunoassay; transferring; α2,6-sialic acid