Determination of Albumin Concentration by MIP QCM sensor

Outline Introduction Experimental Results and discussion Conclusions

Introduction 1.Albumin is a major plasma protein in the urine or blood which is an indication of urine or blood which is an indication of some types of kidney or liver dysfunction. some types of kidney or liver dysfunction. 2.The common method to determine the albumin by dye-binding method. albumin by dye-binding method. ex.BCG( bromocresol green) or ex.BCG( bromocresol green) or BCP(bromocresol purple). BCP(bromocresol purple).

3.However, BCG and BCP methods have poor selectivity and sensitivity,when the poor selectivity and sensitivity,when the specimen is a protein mixture or less specimen is a protein mixture or less amount. amount. 4.The molecular imprinting polymer has high selectivity and high sensitivity recognition selectivity and high sensitivity recognition site which due to the shape of molecular site which due to the shape of molecular conformation and functional group interaction conformation and functional group interaction of target and monomer molecules. of target and monomer molecules.

5.QCM has many applications, especially on the trace analysis. on the trace analysis.

Property of Albumin Half-life: 17 to 23 day Molecular weight: 68,000 Isoelectric point: 4.8 Concentration of human serum: 3.0 to 5.0 g/dl

Experimental

Fabrication Sequence of MIP powder Preparation of MIPcrush of MIP Screening of MIP Washing with 20 wt.% methanol and drying of MIP

Fabrication Sequence of MIP thin film Prepare polymerization reaction solution Extend by spin coating Washing with 20 wt.% methanol and drying MIP Determine adsorption amount by QCM system

Results and discussion

HPLC analysis of protein mixture

HPLC analysis of protein mixture solution before of adsorption

HPLC analysis of protein mixture solution After adsorption of MIP

HPLC analysis of protein mixture solution After adsorption of non-MIP

The Amount of Adsorption for MIP by Using HPLC Cytochrome CLysozymeAlbuminMyoglobin M.W Residence time(min) Adsorption amount per gram MIP 5.16* * * *10 -4 Adsorption mass ratio (MIP) Adsorption mole ratio (MIP)

The Amount of Adsorption for non-MIP determined by HPLC Cytochrome CLysozymeAlbuminMyoglobin Adsorption amount per gram non-MIP 7.61* * * *10 -3 Adsorption mass ratio (non-MIP) Adsorption mole ratio (non-MIP)

Calculate Adsorption amount of Albumin on the QCM ∆F=Cf o 2 ∆M/A+Cf o 3/2 (ρη) 1/2 ∆F: frequency of quartz crystal chip change amount. F 0 : initial frequency of quartz crystal chip. C : quartz material constant (-2.26*10 -6 cm 2 /Hz*g ). ∆M: mass of absorption on the quartz crystal chip. A: surface area of chip.  : system solution density (g/cm 3 ).  : system solution viscosity (g/cm*s)

Loading curve of Quartz Crystal Microbalance Au-NH 2 electrode

Loading curve of Quartz Crystal Microbalance Au-COOH electrode

Loading curve of Quartz Crystal Microbalance Au-OH electrode

Loading curve of Quartz Crystal Microbalance Au electrode

The Amount of Adsorption for MIP by Using HPLC Sample name1234 Functional group-COOHNH 2 OH Steady state time (min) Mass of coating (g) 1.79* * * *10 -7 Adsorption amount (g) 7.10* * * *10 -8 Adsorption amount (g) per gram MIP

Conclusions 1.The prepared MIP can obtain high selectivity and high adsorption capacity for Albumin. 2. The optima QCM electrode is Au-OH QCM electrode for this study.

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