Determination of Albumin Determination of Albumin Concentration by MIP QCM sensorConcentration by MIP QCM sensor

OutlineOutline

• Introduction

• Experimental

• Results and discussion

• Conclusions

IntroductionIntroduction

1.Albumin is a major plasma protein in the1.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 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).

33.However, BCG and BCP methods have.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 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 interactionconformation and functional group interaction of target and monomer molecules.of target and monomer molecules.

5.QCM has many applications, especially 5.QCM has many applications, especially

on the trace analysis.on the trace analysis.

Property of AlbuminProperty 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

ExperimentalExperimental

Fabrication Sequence of MIP powderFabrication Sequence of MIP powder

Preparation of MIP crush of MIP

Screening of MIPWashing with 20 wt.%

methanol and drying of

MIP

Fabrication Sequence of MIP thin filmFabrication Sequence of MIP thin film

Prepare polymerization

reaction solutionExtend by spin coating

Washing with 20 wt.%

methanol and drying MIP

Determine adsorption

amount by QCM system

Results and discussionResults and discussion

HPLC analysis of protein mixtureHPLC analysis of protein mixture

minutes

0 5 10 15 20 25 30

mV

0

20

40

60

80

100

1: Cytochrome c2: Lysozyme3: Albumimn4: Myoglobin

1.

2.

3.

4.

HPLC analysis of pHPLC analysis of protein mixture solutionrotein mixture solution

minutes

0 5 10 15 20 25 30

mV

0

20

40

60

80

100

1.

2.

3.

4.

before of adsorption

HPLC analysis of pHPLC analysis of protein mixture solutionrotein mixture solution

minutes

0 5 10 15 20 25 30 35

mV

0

20

40

60

80

100

1.

2.

4.

After adsorption of MIP

HPLC analysis of pHPLC analysis of protein mixture solutionrotein mixture solution

minutes

0 5 10 15 20 25 30

mV

0

10

20

30

40

50

60

70

80

90

100

1.

2.

3.

4.

After adsorption of non-MIP

The Amount of Adsorption for MIP by Using HPLCThe Amount of Adsorption for MIP by Using HPLC

Cytochrome C Lysozyme Albumin Myoglobin

M.W 12327 14300 68000 17000

Residence time(min)

18.9 20.5 22.2 24.0

Adsorption amount per gram MIP

5.16*10-4 2.08*10-4 9*10-3 6.90*10-4

Adsorption mass ratio (MIP)

2.5 1 43.3 3.3

Adsorption mole ratio (MIP)

2.9 1 9.1 2.8

The Amount of Adsorption for non-MIP The Amount of Adsorption for non-MIP determined by HPLCdetermined by HPLC

Cytochrome C Lysozyme Albumin Myoglobin

Adsorption amount per

gram non-MIP7.61*10-4 2.69*10-3 5.82*10-3 1.62*10-3

Adsorption mass ratio (non-MIP)

1 3.5 7.7 2.1

Adsorption mole ratio (non-MIP)

1 3.1 1.4 1.5

Calculate Adsorption amount of Albumin on the Calculate Adsorption amount of Albumin on the QCMQCM

∆F=Cfo2 ∆M/A+Cfo

3/2(ρη)1/2

∆F: frequency of quartz crystal chip change amount. F0: initial frequency of quartz crystal chip. C : quartz material constant (-2.26*10-6 cm2/Hz*g ).∆M: mass of absorption on the quartz crystal chip. A: surface area of chip. : system solution density (g/cm3). : system solution viscosity (g/cm*s)

Loading curve of Quartz Crystal MicrobalanceLoading curve of Quartz Crystal Microbalance

0 500 1000-160

-140

-120

-100

-80

-60

-40

-20

0

20

40

60fr

eque

ncy

(Hz)

time (sec)

Au-NH2 electrode

Loading curve of Quartz Crystal MicrobalanceLoading curve of Quartz Crystal Microbalance

0 1000 2000

-200

-150

-100

-50

0

freq

uenc

y (H

z)

time (sec)

Au-COOH electrode

Loading curve of Quartz Crystal MicrobalanceLoading curve of Quartz Crystal Microbalance

0 200 400 600 800 1000 1200 1400

-500

-400

-300

-200

-100

0

freq

uenc

y (H

z)

time (sec)

Au-OH electrode

Loading curve of Quartz Crystal MicrobalanceLoading curve of Quartz Crystal Microbalance

0 500 1000 1500 2000 2500-200

-150

-100

-50

0

freq

uenc

y (H

z)

time (sec)

Au electrode

The Amount of Adsorption for MIP by Using The Amount of Adsorption for MIP by Using HPLCHPLC

Sample name 1 2 3 4

Functional group - COOH NH2 OH

Steady state time (min) 33 27 15 19

Mass of coating (g) 1.79*10-6 1.04*10-6 7.04*10-7 3.13*10-7

Adsorption amount (g) 7.10*10-8 7.95*10-8 5.14*10-8 2.86*10-8

Adsorption amount (g) per gram MIP

0.04 0.08 0.07 0.09

ConclusionsConclusions

1.1. The prepared MIP can obtain high selectivity and The prepared MIP can obtain high selectivity and

high adsorption capacity for Albumin .high adsorption capacity for Albumin .

22. The optima QCM electrode is Au-OH QCM . The optima QCM electrode is Au-OH QCM

electrode for this study.electrode for this study.

Thank you for your attentionThank you for your attention