Limits...
Assay of hemoglobin A 1c using lectin from Aleuria aurantia

View Article: PubMed Central - PubMed

ABSTRACT

Hemoglobin A1c (HbA1c) 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 HbA1c to total Hb is an important biomarker for the diagnosis of diabetes. HbA1c-binding lectins were screened from 29 sources of lectin, and the lectin from Aleuria aurantia (AAL) was revealed to have higher affinity to HbA1c than to Hb. The concentration of HbA1c 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 HbA1c with an anti-HbA1c antibody inhibited the binding, suggesting that AAL binds to the N-terminal fructosyl valine epitope of HbA1c. Higher inhibitory effect was observed for 10 mM d-fructose than for the same concentrations of l-fucose, d-fucose, or d-glucose.

No MeSH data available.


Related in: MedlinePlus

Inhibition of the binding by the monoclonal antibody against HbA1c. The lectin-based ELISA was performed in the presence (open triangle) and the absence (closed circle) of the anti-HbA1c monoclonal antibody (10 μg/ml)
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC5120163&req=5

Fig3: Inhibition of the binding by the monoclonal antibody against HbA1c. The lectin-based ELISA was performed in the presence (open triangle) and the absence (closed circle) of the anti-HbA1c monoclonal antibody (10 μg/ml)

Mentions: The inhibitory effect of the monoclonal antibody against the N-terminal fructosyl valine of β-chain of HbA1c on the binding between AAL and HbA1c was assayed in order to obtain information about the binding position of HbA1c. The antibody was incubated with denatured HbA1c and lectin-based ELISA was performed. Figure 3 shows the effect of the antibody on the binding. The binding was inhibited by the incubation of the antibody and HbA1c prior to the lectin-based ELISA. This result suggests that AAL binds to the N-terminal fructosyl valine of the β-chain of HbA1c.Fig. 3


Assay of hemoglobin A 1c using lectin from Aleuria aurantia
Inhibition of the binding by the monoclonal antibody against HbA1c. The lectin-based ELISA was performed in the presence (open triangle) and the absence (closed circle) of the anti-HbA1c monoclonal antibody (10 μg/ml)
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC5120163&req=5

Fig3: Inhibition of the binding by the monoclonal antibody against HbA1c. The lectin-based ELISA was performed in the presence (open triangle) and the absence (closed circle) of the anti-HbA1c monoclonal antibody (10 μg/ml)
Mentions: The inhibitory effect of the monoclonal antibody against the N-terminal fructosyl valine of β-chain of HbA1c on the binding between AAL and HbA1c was assayed in order to obtain information about the binding position of HbA1c. The antibody was incubated with denatured HbA1c and lectin-based ELISA was performed. Figure 3 shows the effect of the antibody on the binding. The binding was inhibited by the incubation of the antibody and HbA1c prior to the lectin-based ELISA. This result suggests that AAL binds to the N-terminal fructosyl valine of the β-chain of HbA1c.Fig. 3

View Article: PubMed Central - PubMed

ABSTRACT

Hemoglobin A1c (HbA1c) 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 HbA1c to total Hb is an important biomarker for the diagnosis of diabetes. HbA1c-binding lectins were screened from 29 sources of lectin, and the lectin from Aleuria aurantia (AAL) was revealed to have higher affinity to HbA1c than to Hb. The concentration of HbA1c 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 HbA1c with an anti-HbA1c antibody inhibited the binding, suggesting that AAL binds to the N-terminal fructosyl valine epitope of HbA1c. Higher inhibitory effect was observed for 10 mM d-fructose than for the same concentrations of l-fucose, d-fucose, or d-glucose.

No MeSH data available.


Related in: MedlinePlus