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Measurement and Characterization of the Agglomeration of Multimeric Proteins on Solid-liquid Interface

多聚亚基蛋白质在固-液界面团聚的测量与表征



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ChineseJournalofBioprocessEngineering
Vol.10No.1
Jan.2012
doi:10.3969/j.issn.1672-3678.2012.01.010
OPQI
:2011-07-23
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ËDÙ

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(DPI)
w4Í6B­J
(AFM)
í
î¶K ÈL©¾³MNÓË

ÝÞïð

.ÈÈ¢OÎÿ^Á.³IJüýË
108mg/mL
â

PÚ¶Q:¼ß
©MN³ÕORýË
354nm,
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60~150nm,
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Xyÿ^Á.¶K ÈL
©¾ï
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HBsAg
IJüýË
067mg/mL
â

PÚ¶Q:¼ß©7
DEAE
Zc1ß©MN³ÕORýÄ

106
w
2369nm,
ß©MN³Á.S¯T#ÄmË
288~401
w
520~971nm,
[·ÄÍ
HBsAg
VW³
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:1672-3678(2012)01-0051-07
Measurementandcharacterizationofagglomerationof
multimericproteinsonsolidliquidinterface
DUXiaonan1,2,LIYan1,SUNLijing1,ZHANGYan1,MAGuanghui1,SUZhiguo1,GAOFei1
(1.TheNationalKeyLaboratoryofBiochemicalEngineering,InstituteofProcessEngineering,
ChineseAcademyofSciences,Beijing100190,China;
2.TheGraduateUniversityofChineseAcademyofSciences,Beijing100049,China)
Abstract:Usingtwomultimericproteins,i.e.,hemoglobinandhepatitisBsurfaceantigen(HBsAg),as
modelproteins,theadsorptionbehaviorsonsolidliquidinterfacewereanalyzedbydualpolarizationinter
ferometry(DPI)andatomicforcemicroscope.Itwasshownthattheaveragethicknessofhemoglobinat
tachedtounmodifiedDPIsensorchipwas354nmandthediametersofproteinparticleswere60nmto
150nmat108mg/mL,anditwasfarlargerthanthesizeofsinglehemoglobinmolecular,thusindica
tingthathemoglobinwasagglomeratedonSi—OHsurface.Undertheconcentrationof067mg/mL,the
layerthicknessesofadsorbedHBsAgwas104nmand2369nmonunmodifiedsurfaceandDEAEagar
osesurfacerespectively,andthecorespondingdiameterswere288nmto401nmand520nmto971
nm,itwas131-441foldofsingleHBsAgmolecule.Theresultshowedthatbigagglomerateswere
formedonbothsurfaces.
Keywords:atomicforcemicroscope;dualpolarizationinterferometry;hemoglobin;hepatitisB
surfaceantigen
  
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[1-2]、BSA[3-4]、
{
êh
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¿Àr´Â|IX-

bþÂI

7–+
yzñíšã²‘A’69ö:x[º

›Æƒ
Ãi+8ä
(hepatitisBsurfaceantigen,HBsAg)
Õ
8
100
CôQ|úE

ò@p­’™<¿
20×
106
Eqg‘A’

˕rEp­eé“
22nm
4
5
[6]。
R"÷±­1…­q.‰-€^
HBsAg
<
¿E:;Ÿa=:?

£Ü
50%)。
‘A’–+ÂIE³´Í.T&ö€-.
、¯
MÍ.€m+ƒ€.þ
[7-9]。
ïVPo¬i›U
B

$àÏ<í+Eyz¡ë

(³ŽHPC¬’
€UBË_ÌÍúí³´‘A’–+ñíE8
9
[10]。
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(dualpolarizationinter
ferometry,DPI)
ÕWGE˜™–+ñíEpq’
€

â÷Ï3CëE–+­˜ˆH

c’™
Á
[11]。
䭊ýµ+
(atomicforcemicroscope,
AFM)
âeR~˜–+Eµ~ùȁ‘A’Ep
Û
[12],
ú$$чE•òÓÔZmòCÂ6“
” •–+|yz‘A’Eä{²˜

R"Y

¬
i›ˆ÷³P2!/í+¢IàÆ‘A’“–
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˕ax
[13-14]
þÍ+

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Ç;4ìEQW|

³P
DPI
š
AFM,
³´ÕQE
…Y‘A“¦ŒQi+Eyz&½

©½oD“¦Œ
Qi+J­
DEAE
ö{D

Z^±­1…­qE6’
i+

³´6CQE
HBsAg
“¦ŒQi+
DEAE
ö{Di+Eyzñí

x¿i›

¬CQ‘
A’“” •–+|Eyz™šyzùN

÷#íC
-­q¿ÀrEµ~Xé³´2G?Il1

1 
vw%xy
11 
€|}
Analight 4Ds&„èŒTFpq’,ÕJv
u
Farfield
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Vecco
vw

12 
€vw
хY‘A8@AғC3ãJхY<=
r23
[15];
KLˆœ
(Hansenularpolymorpha)
ij
EƒÃi+8ä8ªf€mvw2G

ò@p­’
™í
3×106;DEAE
ö{DíJ¹

7MÒP2 ˆ
ípq‰

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Qplus
<‰çX

tu
Milipore
vw

¹º
;DPI
²˜MÔÕJvu
Farfield
vw

13 
€xy
131 DEAE
ö{DE¹º
ž3
5g
’™p?í
2%
Eö{DÞ˕

N
¿
10mL25mol/L
E
NaOH
˕Ñ
,40℃
klÀ
¨-
40min。
ÐVÊ
15mol/LDEAEHCl
ë*
4h,
¹<
DEAE
ö{D˕

òqx§VÊ
05
mol/LNaOH
˕

H
DEAE
ö{Di+ßU—V

Æòq
6h。
DEAE
îQcE²˜

—BNµ.²˜îQ


P
HCl
rs˕Nµ
DEAE
ö{D˕

Û
IÒP
HCl
²Ì5áîQEc
(LD),
LD=cHClVHCl/Vgel (1)
132 
÷
DPI
C3²˜‘A’Eyz
m
1 DPI
ɺ}“
Fig.1 SchematicrepresentationofDPIsensorchip
DPI
C3ˆ“¦ŒQMÔ|¾Ñ

ÒP˕ˆ
í<‰çî¹

©_
01μmG¿û。VüpH74,
20mmol/LPB
íJû•

t—POºxº
15~20
min;
þ¡½«&

2«‡ÛÜxº

¡

…Y‘A

HBsAg
E’™£pÇí
108、067mg/mL,
ö{
D’™£í
03mg/mL。
DPI
²˜MÔi+ö&xS

pÇíⅠxS
ⅡxS(Ö1)。ÆÖ1Òr:ⅠxSí¦ŒQi+,
YⅡxS†Pú DEAEö{Di+,ÐV“7i+
yz‘A’

÷
20mmol/LPB
í ãò

MÔP
80%
ƒw<‰ç½ñQØQs

2«Ë•÷
60
μL/minE=½«2min,Æ÷20μL/minE 
25
€
 
m
 

 
4
 
¿
 
À
  
!
10
"
 
½«
3min。
P
DPI
?Ièé\l5áMÔi+‘
A’EyzH

c’™

133 
÷
AFM
~&–+‘AEùÈ
Y
132
ri+yz‘A’E
DPI
MÔÈÜJ
û•r

HP
AFM
•²RSZmÓÔMÔE&x
S

²˜” •–+yz‘AEùÈ°›


ver
tical

horizontal
>?“

45
,sum
Áí

45

Ó
ÔÏÐí
5~50μm。x¿AFM\l5á‘A’Q
R

©Ï<–+|yz‘A’Eg‹ÖS

2 
‚ƒ%„)
m
2 
*?sÁÕÊËK®W‰†Ì
Fig.2 AdsorptionofhemoglobinonSi—OHsensorchip
surfaceasshownby(a)themeasuredTM and
TEphasechanges,(b)theresolvedlayer
thickness,densityandmassloadingafter
hemoglobininjection
21 DPI
%
AFM
ÍÜ®œÊËK®W†Ì‰*
?sÁ
  108mg/mL
…Y‘A˕ ¿
DPI
E¦ŒQ
MÔi+
,DPI
x¿ßÎTU
(transversemagnetic,
TM)
ßΗU
(transverseelectronic,TE)
&„èŒ
²˜i+yzñí
,TM、TE
¡YX-ÆÖ
2(a)
Ò
r


2(a)
âï

…Y‘A½«V
TM、TE
¡Y
ôý7]

nô‘A’“i+|öôýEyz

…
Y‘A½«V

¦ŒQi+yz­EH

‚M×
Ø
)、
ã{’™

™^×Ø

c

JM×Ø

EX
-ÆÖ
2(b)
Òr


2(b)
âï

yzH’
™ôý|7V8ÜLµ

iô…Y‘A“i+Ey
zj¿˜?

c—|7VÀ,

iô‘A’­8
[©ÌÍX
“¦ŒQi+ùúLµE‘


‘A’“¦ŒQi+Eyz­Hyz’
™â8
DPI
\l5á<¿

ÛI‘A’Eyz’™
:â<´ãp­‘A’ÒšE+Ì

Æm
(2)
Òr

A=
Mw
NAmL
(2)
mr
:A
íãp­‘A’Òš+Ì
;Mwí‘A’E
ò@p­’™
;NAítVWÞ¶ð?;mLíyz
’™

…Y‘Ap­QRí
64nm×55nm×50
nm[5]。
8i

âï

8
DPI
²˜¿E–+yz‘A
Hí
354nm,
?܅Y‘Ap­Ers>?

5
á<¿Eãp­‘AҚ+Ìí
13010nm2,

0ð|IE…Y‘AҚIE¶x+Ì
(3520
nm2[5])。
nô…Y‘A“” •–+|E|I‡€
6X-

â!Xú6X˜ùN

®
1 
ÊËK®W*?sÁ‰†ÌÎÏ
Table1 AmountofAbsorbedhemoglobinon
Si—OHsurface


nm
ã{’™

(ng·mm-2)
ãp­‘AҚ+Ì

nm2
354 082 13010
 
  
í½oD6‚
DPI
MÔi+yz…Y‘AE&
½

“¦ŒQMÔi+yz…Y‘AV

eRHP
AFM
Çä“Ë•axrÓÔ

CÂ6
AFM
@
DPI
MÔEä{²˜

êë
AFM
E•òÓÔZm>ï‡
6yzE…Y‘Aoeèܔ •–+|

¦ŒQi+…Y‘AE
AFM
ÓÔ/æÆÖ

Òr

Ö
3(a)
í
AFM
ÓÔ]Ö

Ö
3(b)
íò*
E‘A’g‹Ö

Ör0^Yètii+yzE…
Y‘A

_
AFM
\l˜™

…Y‘AYèeéí
60~150nm,
ž]Üp­QR
(64nm×55nm×
50nm),
R"yz“” •–+|E…Y‘AÕ÷
x—²EùN+“
。DPI
²˜¿E‘Ayz­H
?Ü7p­QR
,AFM
²˜¿…Y‘AYèeé
ž]Ü7p­QR

x¿’€E³Ži›nô

¦
ŒQi+|yzE…Y‘AX
©“–+
µaxEF)À

‘Ap­9´Â6—

35 
!

# ”{»þ

CQ‘A’“” •–+—E˜™ši›
m3 
ÊËK®W†Ì*?sÁ‰
AFM
ÐÑm

ÐÑâÒ
5μm)
Fig.3 AFM heightand3Dimageofhemoglobinabsorbed
onSi—OHsurface(scannedarea5μm)
22 DPI
%
AFM
ÍÜ®œÊËK®W†Ì‰¢
Ó®WdÌ
  
½oD%&/|QqgEƒÃi+8ä
(HBsAg)
“¦ŒQi+Eyz&½
。067mg/mL
HBsAg
˕ ¿
DPI
E¦ŒQMÔi+

½«‘A
’6V

MÔi+E
TM、TE
¡YX-ÆÖ
4(a)
Ò
r


4(a)
âï
:HBsAg
½«V
,TM

TE
¡Y
ôý7]


HBsAg
“¦ŒQi+j¿oµ™E
yz

¦ŒQi+‘Ayz­H

‚M×Ø
)、
c


JM×Ø

yzã{’™

™^×Ø

EX-
89ÆÖ
4(b)
Òr


4(b)
âï

‘Ayz­
Hyz’™—|7V8ÜLµ


HBsAg
“
¦ŒQi+Eyz™ÌÍLµ

yz­c|7ó
˜?

iô‘A’“i+ùúLµEyz­

HBsAg
“¦ŒQi+Eyz™Æi

Òr

8
i

âï
:DPI
²˜¿E‘A­Hí
106nm,
ž

HBsAg
“Ë•rEeé
22nm[5],
(ãp­‘
AҚ+Ì
1488095nm2
ž]Ü0ð|IE
HB
sAg
ҚIE+Ì
484nm2[5],
nôi+yzE
HB
sAg
|I¹êÜ˕axrE|I

â!X
m
4 
¢Ó®WdÌÕÊËK®W‰†Ì
Fig.4 AdsorptionofHBsAgonSi—OHsensorchip
surface
®
2 HBsAg
ÕÊËK®W‰†ÌÎÏ
Table2 AmountofabsorbedHBsAgonSi—OHsurface


nm
ã{’™

(ng·mm-2)
ãp­‘AҚ+Ì

nm2
106 034 1488095
 
HBsAg
“¦ŒQi+yzV

P
AFM
•òÓÔ
ZmeRÓÔ
DPI


Ï<¦ŒQi+|yz
HB
sAg
EùÈ

/æ‹Ö
5。
Ö
5(a)
í
AFM
ÓÔ]
Ö

Ö
5(b)
íò*E‘AYèg‹Ö

_
AFM
\l
˜™i+yzE
HBsAg


‡Âeéí
288~401
nm,
ž]Ü˕rEeé
(22nm)[5];
¦ŒQi+y
z‘AEãôYèҚ+Ìí
65144~505170nm2,
>]Ü
DPI
²˜¿Eãp­‘AҚ+Ì
1488095nm2。
x¿
AFM

DPI
E³Ži›

‡Â
“” •–+yzE
HBsAg
Õ÷—²Eùm+“

23 DPI
%
AFM
ÍÜ®œ
DEAE
Ô!Ä®W†
̉¢Ó®WdÌ
  HBsAg
“±­1…­qrE:;Ÿa=?Ü
50%,
Õ3ô‰-45E`Z

R"í2]a=

³
´6
HBsAg
“±­1…­q6’i+Eyz&½

45
€
 
m
 

 
4
 
¿
 
À
  
!
10
"
 
m5 
ÊËK®W†Ì¢Ó®Wd̉
AFM
ÐÑm

ÐÑâÒ
10μm)
Fig.5 AFM heightand3DimageofHBsAgabsorbedon
SiOHsurface(scannedarea10μm)
DEAE
ö{DÕðPE‰-
HBsAg
E±­1…6
’

77µ_QR:x

ûöqgEÑLÜ9Û
û/|

‚.CÂn§ÑLE“Ø~&

R"“Œ
QMÔi+J­
DEAE
ö{Dp­$Z^±­1…
­qE6’i+

x¿
DPI
“Øi›
HBsAg
š
DE
AE
ö{Di+EòwìP

Ö
6(a)
í
TE、TM
¡YX-Ö

ÕÖ
6(a)
â
÷m´

½«

¡
DEAE
ö{D˕6V
,TE、TM
ÁöoµE7]


DEAE
ö{D“¦ŒQi+
ùúyz­

R)½«
067mg/mLHBsAg
˕

TE、TM
¡Yôý7]

nô‘A’“
DEAE
ö{D
i+öx™yz

Ö
6(b)
ýr6i+yz­EH


‚M×Ø
)、


JM×Ø

ã{’™

™^
×Ø

X-89

½«
DEAE
ö{D6V



H
’™ö7]

nô“i+j¿6oµ™Ey
z

“
DEAE
ö{Di+|

‘A­EH’™Ì
͞


HBsAg
ê«öyz

(cE·£i
ô‘A­Ã
DEAE
ö{D­[©

m
6 HBsAg
Õ
DEAE
Ô!Ä®W‰†Ì
Fig.6 AdsorptionofHBsAgonDEAEagarosesurface
asshownby(a)themeasuredTM andTE
phasechanges,(b)theresolvedlayer
thickness,densityandmassloading
aftersampleinjection
HBsAg
“
DEAE
ö{Di+Eyz™Æi

Ò
r

8i

âï
:DEAE
ö{Di+‘Ayz­EH
âj¿
2369nm,
]ܦŒQi+E‘A­H
(106nm),DEAE
ö{Di+
HBsAg
Eyz’™í
777ng/mm2,
]ܦŒQi+E
034ng/mm2。
HBsAg
þ—ví
46,
“
pH
í
74
EJû•ÑßU
—V

(
DEAE
ö{Di+ß0—

—VìPBC
HBsAg
“
DEAE
ö{Di+öQ]Eyz™

DEAE
ö{Di+‘AEyz­H
(2369
nm)
š
HBsAg
E˕reé
(22nm)


ãp­
Қ+Ìí
64292nm2(2533nm×2533nm),
š
‘A’Ë•rEeé
(22nm)



HBsAg
“
DEAE
ö{Di+E|IšË•rE|Iòú

®
3 DEAE
Ô!Ä®W
HBsAg
‰†ÌÎÏ
Table3 AmountofabsorbedHBsAgonDEAE
agarosesurface


nm
ã{’™

(ng·mm-2)
ãp­‘AҚ+Ì

nm2
2369 777 64292
55 
!

# ”{»þ

CQ‘A’“” •–+—E˜™ši›
  AFM
“•òaxÀÓÔ
DEAE
ö{Di+yz
E
HBsAg,
/æ‹Ö
7。
Ö
7(a)
í‘A’E]Ö

Ö
7(b)
í@*‘A’YèEg‹Ö



âï

_
AFM
\l˜™
DEAE
ö{Di+E‘AYèeé
“
520~971nm
69

]ܦŒQi+‘AYèEe
é
(288~401nm),
” •–+|E‘AYèe鈞

HBsAg
˕axrEeé
(22nm)。DEAE
ö
{Di+yz‘AEãôYèҚ+Ìí
212317~
740505nm2,

DPI
²˜Eãp­‘AҚ+Ì
64292nm2。
x¿’€E³Ži›‡Â” •–+|
E
HBsAg
ùú6—

©“
DEAE
ö{Di+E
—ÂIÓ¦ŒQi+|EQý;

m
7 DEAE
Ô!Ä®W†Ì¢Ó®Wd̉
AFM
ÐÑm

ÐÑâÒ
15μm)
Fig.7 AFM heightand3DimageofHBsAgabsorbedon
DEAEagarosesurface(scannedarea15μm)
“¦ŒQi+
DEAE
ö{Di+yzE
HB
sAg
E
AFM
ÓÔÖS

ÆÖ

Òr

Ö
8(a)
ir¦
ŒQi+yzE
HBsAg,
Ö
8(b)
ir
DEAE
ö{D
i+yzE
HBsAg。
x¿@Ãâ÷‡Â

¦ŒQi
+
HBsAg
Eyz™¼ž1Ü
DEAE
ö{Di+E
yz™
,DPI
²˜¿E¦ŒQi+
HBsAg
yz™í
034ng/mm2,DEAE
ö{Di+í
778ng/mm2,
DPI
²˜/æš
AFM
ÓÔ/æò˜

m
8 
ÊËK®W
(a)

DEAE
Ô!Ä®W
(b)
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[1] SuTJ,GreenRJ,WangY,etal.Adsorptionoflysozymeontothe
siliconoxidesurfacechemicalygraftedwithamonolayerofpenta
decyl1ol[J].Langmuir,2000,16(11):49995007.
[2] LuJR,SwannMJ,PeelLL,etal.Lysozymeadsorptionstudies
atthesilica/waterinterfaceusingdualpolarizationinterferometry
[J].Langmuir,2004,20(5):18271832.
[3] BiesheuvelPM,WitemannA.Amodifiedboxmodelincluding
chargeregulationforproteinadsorptioninasphericalpolyelectro
lytebrush[J].JPhysChemB,2005,109(9):42094214.
[4] SonessonAW,CalisenTH,BrismarH,etal.Adsorptionandac
tivityofThermomyceslanuginosuslipaseonhydrophobicandhy
drophilicsurfacesmeasuredwithdualpolarizationinterferometry
(DPI)andconfocalmicroscopy[J].ColoidSurfaceB,2008,61
(2):208215.
[5] BrussV,GerhardtE,VielufK,etal.Functionsofthelargehepa
titisBvirussurfaceproteininviralparticlemorphogenesis[J].
Intervirology,1996,39(1/2):2331.
[6] HeitzF,VanMauN.Proteinstructuralchangesinducedbytheirup
takeatinterfaces[J].BiochimBiophysActa,2002,1597(1):111.
[7] CooperM A.Biosensorprofilingofmolecularinteractionsin
pharmacology[J].CurOpinPharmacol,2003,3(5):557562.
[8] PanceraSM,GliemannH,SchimmelT,etal.EfectofpHonthe
adsorptionandactivityofcreatinephosphokinase[J].JPhys
ChemB,2006,110(6):26742680.
[9] ZhouWB,BiJX,JansoJC,etal.Ionexchangechromatography
ofhepatitisBvirussurfaceantigenfromarecombinantChinese
hamsterovarycelline[J].JChromatogrA,2005,1095(1/2):
119125.
[10] HuangYD,BiJX,ZhouWB,etal.Improvingrecoveryofre
combinanthepatitisBvirussurfaceantigenbyionexchangechro
matographicsupportswithlowliganddensity[J].ProcessBio
chem,2006,41(11):23202326.
[11] XuY,TakaiM,IshiharaK.Proteinadsorptionandceladhesion
oncationic,neutral,andanionic2methacryloyloxyethylphospho
rylcholinecopolymersurfaces[J].Biomaterials,2009,30(28):
49304938.
[12] SwannMJ,PeelLL,CaringtonS,etal.Dualpolarizationinter
ferometry:ananalyticaltechniquetomeasurechangesinprotein
structureinrealtime,todeterminethestoichiometryofbindinge
vents,andtodiferentiatebetweenspecificandnonspecificinter
actions[J].AnalBiochem,2004,329(2):190198.
[13] LinSM,LeeCK,HsuSM,etal.Measurementofdimensionsof
pentagonaldoughnutshapedCreactiveproteinusinganatomic
forcemicroscopeandadualpolarisationinterferometricbiosensor
[J].BiosensBioelectron,2006,22(2):323327.
[14] HuT,SuZG.Bovineserumalbuminbovinehemoglobinconju
gateasacandidatebloodsubstitute[J].BiotechnolLet,2002,24
(4):275278.
[15] 
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