albumin mip prepared in aqueous solution system

Albumin MIP Prepared in Aqueous Albumin MIP Prepared in Aqueous Solution SystemSolution System

Speaker : Thong-Yueh LinSpeaker : Thong-Yueh Lin

報告者報告者 : : 林聰樂林聰樂

OutlineOutline

• Introduction

• Experimental

• Results and discussion

• Conclusions

IntroductionIntroduction

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

MaterialsMaterials

MonomerMonomer

3-dimethylaminopropyl metacrlamide:3-dimethylaminopropyl metacrlamide:DMAPMADMAPMA

acrylamide:AMacrylamide:AM

Allylamine:AAAllylamine:AA

Methacrylic acid:MAAMethacrylic acid:MAA

Hydroxy ethyl methacrylate:HEMAHydroxy ethyl methacrylate:HEMA

Crosslinking agentCrosslinking agent

Tetraethylene glycol dimethaTetraethylene glycol dimethacrylate:TEGDMAcrylate:TEGDMA

Polyethylene glycol dimethaPolyethylene glycol dimethacrylate:PEGDMAcrylate:PEGDMA

MaterialsMaterials

InitiatorsInitiators

Ammonium persulfate:APSAmmonium persulfate:APS

Cumene hydroperoxide:CHPCumene hydroperoxide:CHP

Tetramethyl ethylenediamine:TMEDATetramethyl ethylenediamine:TMEDA

MaterialsMaterials

Fabrication Sequence of MIPFabrication Sequence of MIP

Preparation of MIP crush of MIP

Screening of MIPWashing with 20 wt.%

methanol and drying of

MIP

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.

Typical calibration curve of Typical calibration curve of Cytochrome CCytochrome C by using HPLC by using HPLC

Concentration of cytochrome C (g/ml)

0.000 0.001 0.002 0.003 0.004 0.005 0.006

Inte

grat

e ar

ea ( v

*sec

)

0

1e+6

2e+6

3e+6

4e+6

5e+6

6e+6

7e+6

b = -115699.1074m= 1208803751r2= 0.9999

Typical calibration curve of Typical calibration curve of AlbuminAlbumin by using HPLC by using HPLC

concentration (g/ml)

0.000 0.001 0.002 0.003 0.004 0.005 0.006

inte

grat

e ar

ea(

v*se

c)

0.0

2.0e+5

4.0e+5

6.0e+5

8.0e+5

1.0e+6

1.2e+6

1.4e+6

1.6e+6

b=15164.4912m=269447958.7r2=0.9967

Typical calibration curve of Typical calibration curve of LysozymeLysozyme by using HPLC by using HPLC

contentration (g/ml)

0.000 0.001 0.002 0.003 0.004 0.005 0.006

inte

grat

e ar

ea ( v

*sec

)

0

2e+6

4e+6

6e+6

8e+6

1e+7

b=104300.9234m=1707906682r2=0.9994

Typical calibration curve of Typical calibration curve of MyoglobinMyoglobin by using HPLC by using HPLC

concentration (g/ml)

0.000 0.001 0.002 0.003 0.004 0.005 0.006

inte

grat

e ar

ea ( v

*sec

)

0

2e+6

4e+6

6e+6

8e+6

b=-53712.5055m=1417736164r2=0.9998

Mas

s ra

tio

of a

dso

rbat

e on

MIP

0

2

4

6

8

10

12

14

16

Cytochrome C Lysozyme Albumin Myoglobin

Selectivity mass ratio to adsorbent of M.I.PSelectivity mass ratio to adsorbent of M.I.P

Sample name: DMAPMA-TEGDMA

Albumin 4.5%

Water 10.4%

DMAPMA 5.0%

TEGDMA 80%

APS,TMEDA

Selectivity:

1:1.9:14.2:1.4


Mas

s ra

tio

of a

dso

rbat

e on

MIP

0

20

40

60

80

100

120

140

160

180


Sample name: MAA-TEGDMA

Albumin 3.1%

Water 7.3%

MAA 2.7%

TEGDMA 86.8%

APS,TMEDA

Selectivity:

52.2:160.1:1:

77.5


Sample name: AM-TEGDMA 1M

ass

rati

o of

ad

sorb

ate

on M

IP

0

1

2

3

4

5

6

7


Albumin 2.4%

Water 9.2%

AM 2.4%

TEGDMA 86%

APS,TMEDA

Selectivity:

1:4.1:6.5:2.5


Sample name: AM-TEGDMA 2M

ass

rati

o of

ad

sorb

ate

on M

IP

0

5

10

15

20

25

30


Albumin 2.4%

Water 11.2%

AM 2.3%

TEGDMA 84%

CHP,TMEDA

Selectivity

4.4:25.8:8.9:1


Sample name: HEMA-PEGDMAM

ass

rati

o of

ad

sorb

ate

on M

IP

0

1

2

3

4

5


Albumin 4.5%

Water 10.5%

HEMA 5.1%

PEGDMA 80%

APS,TMEDA

Selectivity

1: 4.3: 1: 1.8


Sample name: DMAPMA-PEGDMAM

ass

rati

o of

ad

sorb

ate

on M

IP

0

1

2

3

4

5

6

7


Albumin 5.4%

Water 12.6%

DMAPMA 6.0%

PEGDMA 76%

APS,TMEDA

Selectivity

3.1:6.4:5.4:1

Mass ratio of Adsorbent for different composition Mass ratio of Adsorbent for different composition

MIPMIP

Sample name Mass ratio of adsorbent(Cytochrome C: Lysozyme: Albumin: Myoglobin)

DMAPMA-TEGDMA 1 : 1.9 : 14.2 : 1.4MAA-TEGDMA 52.2 : 160.3 : 1 : 77.5AM-TEGDMA 1 : 4.1 : 6.5 : 2.5AM-TEGDMA 4.4 : 25.8 : 8.9 : 1

HEMA-PEGDMA 1 : 4.3 : 1 : 1.8DMAPMA-PEGDMA 3.1 : 6.4 : 5.4 : 1

ConclusionsConclusions

1.1. The DMAPMA-TEGDMA MIP can obtain high The DMAPMA-TEGDMA MIP can obtain high

selectivity for albumin selectivity for albumin

2.The mass ratio of DMAPMA-TEGDMA MIP for adsorbent2.The mass ratio of DMAPMA-TEGDMA MIP for adsorbent

= = Albumin : Lysozyme : Myoglobin : Cytochrome CAlbumin : Lysozyme : Myoglobin : Cytochrome C

= = 14.2 : 1.9 : 1.4 : 114.2 : 1.9 : 1.4 : 1

Thank you for your attentionThank you for your attention

albumin mip prepared in aqueous solution system

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