Assay of hemoglobin A1c using lectin from Aleuria aurantia

Masato Kabata; Erina Hase; Kouta Kimura; Yuka Kobayashi; Yasushi Ueno; Kazuaki Yoshimune

doi:10.1186/s13568-016-0288-7

Assay of hemoglobin A_1c using lectin from Aleuria aurantia

AMB Express. 2016 Dec;6(1):119. doi: 10.1186/s13568-016-0288-7. Epub 2016 Nov 22.

Authors

Masato Kabata¹, Erina Hase², Kouta Kimura^{2

3}, Yuka Kobayashi⁴, Yasushi Ueno⁴, Kazuaki Yoshimune^{5

6}

Affiliations

¹ Department of Applied Molecular Chemistry, Graduate School of Industrial Technology, Nihon University, 1-2-1, Izumichou, Narashino, Chiba, 275-8575, Japan.
² Department of Applied Molecular Chemistry, College of Industrial Technology, Nihon University, 1-2-1, Izumichou, Narashino, Chiba, 275-8575, Japan.
³ Nacalai tesque Inc., Kyoto, Japan.
⁴ Product Development Laboratory, J-oil Mills Inc., 11, Kagetoricho Totsuka-ku, Yokohama, Kanagawa, 245-0064, Japan.
⁵ Department of Applied Molecular Chemistry, Graduate School of Industrial Technology, Nihon University, 1-2-1, Izumichou, Narashino, Chiba, 275-8575, Japan. yoshimune.kazuaki@nihon-u.ac.jp.
⁶ Department of Applied Molecular Chemistry, College of Industrial Technology, Nihon University, 1-2-1, Izumichou, Narashino, Chiba, 275-8575, Japan. yoshimune.kazuaki@nihon-u.ac.jp.

Abstract

Hemoglobin A_1c (HbA_1c) has an N-terminal fructosyl valine on the β-chain, and this modification is caused by the non-enzymatic glycosylation of hemoglobin (Hb). The relative concentration ratio of HbA_1c to total Hb is an important biomarker for the diagnosis of diabetes. HbA_1c-binding lectins were screened from 29 sources of lectin, and the lectin from Aleuria aurantia (AAL) was revealed to have higher affinity to HbA_1c than to Hb. The concentration of HbA_1c was determined by lectin-based enzyme-linked immunosorbent assay (ELISA) using the AAL lectin. Higher reproducibility of the assay was observed at 4 °C than at 25 and 37 °C. This observation is consistent with the known temperature-dependent behavior of lectins. Preincubation of HbA_1c with an anti-HbA_1c antibody inhibited the binding, suggesting that AAL binds to the N-terminal fructosyl valine epitope of HbA_1c. Higher inhibitory effect was observed for 10 mM D-fructose than for the same concentrations of L-fucose, D-fucose, or D-glucose.

Keywords: Aleuria aurantia; Hemoglobin A1c; Lectin; Lectin-based enzyme-linked immunosorbent assay.