Enrichment of Glycoproteins using Nanoscale Chelating Concanavalin A Monolithic Capillary Chromatography

Immobilized lectin chromatography can be employed for glycoprotein enrichment, but commonly used columns have limitations of yield and resolution. To improve efficiency and to make the technique applicable to minimal sample material, we have developed a nanoscale chelating Concanavalin A (Con A) monolithic capillary prepared using GMA-EDMA (glycidyl methacrylate-co-ethylene dimethacrylate) as polymeric support. Con A was immobilized on Cu(II)-charged iminodiacetic acid (IDA) regenerable sorbents by forming a IDA:Cu(II):Con A sandwich affinity structure that has high column capacity, as well as stability. When compared with conventional Con A lectin chromatography, the monolithic capillary enabled the better reproducible detection of over double the number of unique N-glycoproteins in human urine samples. Utility for analysis of minimal biological samples was confirmed by the successful elucidation of glycoprotein profiles in mouse urine samples at the microliter scale. The improved efficiency of the nanoscale monolithic capillary will impact the analysis of glycoproteins in complex biological samples, especially where only limited material may be available

Enrichment of Glycoproteins using Nanoscale Chelating Concanavalin A Monolithic Capillary Chromatography

Immobilized lectin chromatography can be employed for glycoprotein enrichment, but commonly used columns have limitations of yield and resolution. To improve efficiency and to make the technique applicable to minimal sample material, we have developed a nanoscale chelating Concanavalin A (Con A) monolithic capillary prepared using GMA-EDMA (glycidyl methacrylate-co-ethylene dimethacrylate) as polymeric support. Con A was immobilized on Cu(II)-charged iminodiacetic acid (IDA) regenerable sorbents by forming a IDA:Cu(II):Con A sandwich affinity structure that has high column capacity, as well as stability. When compared with conventional Con A lectin chromatography, the monolithic capillary enabled the better reproducible detection of over double the number of unique N-glycoproteins in human urine samples. Utility for analysis of minimal biological samples was confirmed by the successful elucidation of glycoprotein profiles in mouse urine samples at the microliter scale. The improved efficiency of the nanoscale monolithic capillary will impact the analysis of glycoproteins in complex biological samples, especially where only limited material may be available

Enrichment of Glycoproteins using Nanoscale Chelating Concanavalin A Monolithic Capillary Chromatography

Immobilized lectin chromatography can be employed for glycoprotein enrichment, but commonly used columns have limitations of yield and resolution. To improve efficiency and to make the technique applicable to minimal sample material, we have developed a nanoscale chelating Concanavalin A (Con A) monolithic capillary prepared using GMA-EDMA (glycidyl methacrylate-co-ethylene dimethacrylate) as polymeric support. Con A was immobilized on Cu(II)-charged iminodiacetic acid (IDA) regenerable sorbents by forming a IDA:Cu(II):Con A sandwich affinity structure that has high column capacity, as well as stability. When compared with conventional Con A lectin chromatography, the monolithic capillary enabled the better reproducible detection of over double the number of unique N-glycoproteins in human urine samples. Utility for analysis of minimal biological samples was confirmed by the successful elucidation of glycoprotein profiles in mouse urine samples at the microliter scale. The improved efficiency of the nanoscale monolithic capillary will impact the analysis of glycoproteins in complex biological samples, especially where only limited material may be available

Enrichment of Glycoproteins using Nanoscale Chelating Concanavalin A Monolithic Capillary Chromatography

Immobilized lectin chromatography can be employed for glycoprotein enrichment, but commonly used columns have limitations of yield and resolution. To improve efficiency and to make the technique applicable to minimal sample material, we have developed a nanoscale chelating Concanavalin A (Con A) monolithic capillary prepared using GMA-EDMA (glycidyl methacrylate-co-ethylene dimethacrylate) as polymeric support. Con A was immobilized on Cu(II)-charged iminodiacetic acid (IDA) regenerable sorbents by forming a IDA:Cu(II):Con A sandwich affinity structure that has high column capacity, as well as stability. When compared with conventional Con A lectin chromatography, the monolithic capillary enabled the better reproducible detection of over double the number of unique N-glycoproteins in human urine samples. Utility for analysis of minimal biological samples was confirmed by the successful elucidation of glycoprotein profiles in mouse urine samples at the microliter scale. The improved efficiency of the nanoscale monolithic capillary will impact the analysis of glycoproteins in complex biological samples, especially where only limited material may be available

Enrichment of Glycoproteins using Nanoscale Chelating Concanavalin A Monolithic Capillary Chromatography

Immobilized lectin chromatography can be employed for glycoprotein enrichment, but commonly used columns have limitations of yield and resolution. To improve efficiency and to make the technique applicable to minimal sample material, we have developed a nanoscale chelating Concanavalin A (Con A) monolithic capillary prepared using GMA-EDMA (glycidyl methacrylate-co-ethylene dimethacrylate) as polymeric support. Con A was immobilized on Cu(II)-charged iminodiacetic acid (IDA) regenerable sorbents by forming a IDA:Cu(II):Con A sandwich affinity structure that has high column capacity, as well as stability. When compared with conventional Con A lectin chromatography, the monolithic capillary enabled the better reproducible detection of over double the number of unique N-glycoproteins in human urine samples. Utility for analysis of minimal biological samples was confirmed by the successful elucidation of glycoprotein profiles in mouse urine samples at the microliter scale. The improved efficiency of the nanoscale monolithic capillary will impact the analysis of glycoproteins in complex biological samples, especially where only limited material may be available

Enrichment of Glycoproteins using Nanoscale Chelating Concanavalin A Monolithic Capillary Chromatography

Immobilized lectin chromatography can be employed for glycoprotein enrichment, but commonly used columns have limitations of yield and resolution. To improve efficiency and to make the technique applicable to minimal sample material, we have developed a nanoscale chelating Concanavalin A (Con A) monolithic capillary prepared using GMA-EDMA (glycidyl methacrylate-co-ethylene dimethacrylate) as polymeric support. Con A was immobilized on Cu(II)-charged iminodiacetic acid (IDA) regenerable sorbents by forming a IDA:Cu(II):Con A sandwich affinity structure that has high column capacity, as well as stability. When compared with conventional Con A lectin chromatography, the monolithic capillary enabled the better reproducible detection of over double the number of unique N-glycoproteins in human urine samples. Utility for analysis of minimal biological samples was confirmed by the successful elucidation of glycoprotein profiles in mouse urine samples at the microliter scale. The improved efficiency of the nanoscale monolithic capillary will impact the analysis of glycoproteins in complex biological samples, especially where only limited material may be available

Time dependent effect of Diapin on blood glucose level in <i>KKay</i> diabetic mice.

<p>Male <i>KKay</i> diabetic mice were randomly divided into control and treated groups. In control group (•), the mice were fed with regular chaw diet. The mice in the treated groups were fed with regular chow diet containing 6 (Δ) and 12 g/kg Diapin (▪), respectively. The casual blood glucose levels were monitored weekly. n = 10, *<i>p</i><0.05, compared with the control.</p

Effect of Diapin on blood glucose levels in diabetic mice.

<p>After oral loading of Diapin (Δ, 0.5 mg/g; ▪, 1 mg/g) by gavage, OGTT was performed in male (<b>A</b>) <i>ob/ob</i> (n = 10), (<b>B</b>) <i>db/db</i> (n = 10) diabetic mice, (<b>C</b>) <i>KKay</i> (n = 9), and (<b>D</b>) the high fat diet-induced obesity mice (n = 10), in which the wild type male C57BL/6J mice were fed with high fat diet for ten weeks. *<i>p</i><0.05, compared with the controls.</p

Proposed mechanism of action for Diapin to lower blood glucose in T2D.

<p>Diapin is a tripeptide that has potential to treat T2D. Oral administration of Diapin may directly act on enteroendocrine L-cells in gastrointestinal tract to stimulate GLP-1 secretion (dashed line: unknown mechanisms), which in turn stimulates the glucose-dependent insulin secretion. In addition, Diapin in the blood stream may also act on pancreatic β-cells to stimulate insulin secretion (dashed line: unknown mechanisms). This dual effect leads to the elevation of insulin and the effective control of blood glucose in T2D.</p

Effect of Diapin on non-fasting blood glucose levels in <i>KKay</i> diabetic mice.

<p>(A) Male <i>KKay</i> diabetic mice and (<b>B</b>) adult male C57BL/6J mice under non-fasting condition were given either vehicle in control groups or Diapin (1 mg/g bw, n = 9/group) in the treated groups. After Diapin loading, the blood glucose levels were measured every 30 min. *<i>P</i><0.05, compared with the controls.</p