Capillary electrophoresis for protein analysis [Elektronische Ressource] : strategies to prevent protein adsorption and method development using coated capillaries for electrophoresis / von Adhitasari Suratman

Adhitasari

Sujets

• Biologie

Informations

• Publié par: technische_universitat_carolo-wilhelmina_zu_braunschweig
• Publié le: 01 janvier 2008
• Nombre de lectures: 43
• Langue: English
• Poids de l'ouvrage: 1 Mo

Extrait

Capillary Electrophoresis for Protein Analysis

Strategies to Prevent Protein Adsorption
and Method Development Using Coated Capillaries
for Electrophoresis



Von der Fakultät für Lebenswissenschaften
der Technischen Universität Carolo-Wilhelmina
zu Braunschweig


zur Erlangung des Grades einer
Doktorin der Naturwissenschaften
(Dr. rer. nat.)


genehmigte
Dissertation





von Adhitasari Suratman
aus Yogyakarta/ Indonesien























1. Referent: Professor Dr. Hermann Wätzig
2. Referent: Professor Dr. Conrad Kunick
eingereicht am: 27.10.2008
mündliche Prüfung (Disputation) am: 01.12.2008

Druckjahr 2008



VORVERÖFFENTLICHUNGEN

Teilergebnisse aus dieser Arbeit wurden mit Genehmigung der Fakultät für
Lebenswissenschaften, vertreten durch den Mentor der Arbeit, in folgenden Beiträgen vorab
veröffentlicht:

Publikationen
1. Suratman, A., Wätzig, H. Reproducible protein analysis by capillary electrophoresis
using linear polyacrylamide-coated capillaries and hydrochloric acid rinsing.
Electrophoresis 2007, 28, 2324-2328.
2. Suratman, A., Wätzig, H. Long-term precision in capillary isoelectric focusing for
protein analysis. J. Sep. Sci 2008, 31(10), 1834-1840.

Tagungsbeiträge
1. Suratman, A., Graf, M., Prucker, O., Rühe, J., and Wätzig, H. PDMAA Coated
capillaries and trehalose in reducing protein adsorption. The Annual Conference of the
Association of Germany Pharmacists, Marburg, Germany, October 4-7, 2006. (Poster
C099)
2. Suratman, A., Wätzig, H. Reproducible protein analysis by capillary electrophoresis
stusing linear polyacrylamide-coated capillaries and hydrochloric acid rinsing. 31
International Symposium on High Peformance Liquid Phase Separations and Related
Techniques, Ghent, Belgium, June 17-21, 2007. (Poster P18.25)
3. Suratman, A., Wätzig, H., Hydrochloric acid rinsing for obtaining a reproducible
thprotein analysis on CZE and CIEF using linear polyacrylamide-coated capillaries. 12
International meeting on recent developments in pharmaceutical analysis, Island of
Elba, Italy, September 23-26, 2007. (Poster P043)
4. Suratman, A., Wätzig, H. HCl rinsing for obtaining a reproducible protein analysis on
CE using LPA-coated capillaries. The Annual Conference of the Association of
Germany Pharmacists, Erlangen, Germany, October 10-13, 2007. (Poster H12)
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5. Suratman, A., Kühne, S., Wätzig, H., Long-term Precision in Capillary Isoelectric
thFocusing (CIEF) for Protein Analyses. 19 International Symposium on
Pharmaceutical and Biomedical analysis (PBA 2008), Gda ńsk, Poland, June 8-12,
2008. (Poster PA100)
6. Kühne, S., Suratman, A., Wätzig, H., Precision in Capillary Isoelectric Focusing
(CIEF) for Long-term Protein Analyses. The Annual Conference of the Association of
Germany Pharmacists, Bonn, Germany, October 8-11, 2008. (Poster B36)

























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ACKNOWLEDGMENTS

I have the opportunity to express my gratitude to my supervisor Prof. Dr. Hermann Wätzig for
contributed time, constructive discussion, and suggestion during this study.
It is a great pleasure to extend thanks to the members of the examination committee for
constructive comments and reviewing this thesis.
I would like to thank to the member and colleagues of the institute of Pharmaceutical
Chemistry, TU-Braunschweig, especially to Dr. Michael Graf, Dr. Sami El Deeb, Heidi
Köppel, Phillip Hasemann, Simone Schröder, Lukas Kaminski, Sascha Kühne and Xi Deng.
I am deeply thankful to Sascha Kühne for carefully reading this thesis.
I wish to extend my gratitude to the Germany Academic Exchange Service (DAAD) for
financial support.
My grateful thanks are also for Department of Microsystems Engineering (IMTEK), Albert-
Ludwigs Universität Freiburg (Breisgau) for providing polydimethylacrylamide (PDMAA)-
coated capillaries and Polymicro Technologies (Phoenix, AZ, USA) for providing bare fused-
silica and polyacrylamide (LPA)-coated capillaries used in this research.
My grateful thanks to my parents who supported me in all I have done in my life.
Finally, I would like to thank to my husband Atmanto Heru Wibowo and my son Fynn Axel
Wibowo for supporting, encouragement and patience during my study.












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Dedications
to:
My Parents,
Husband, Son and Brother











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List of Abbreviations

abc 4-amino benzoic acid
Bicine N,N-bis(2-hydroxyethyl)glycine
BGE background electrolyte
CAPS 3-(cyclohexylamino)-1-propanesulfonic acid
CD cyclodextrine
CE capillary electrophoresis
CGE gel
CHAPS 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate
CIEF isoelectric focusing
CITP capillary isotachophoresis
CZE zone electrophoresis
DDAB N,N-didodecyl-N,N-dimethylammonium bromide
DEGMA diethylene glycol monomethacrylate
DMAA N,N-dimethylacrylamide
DMSO dimethylsulfoxide
EOF electroosmosis flow
EPDMA epoxy-poly(dimethylacrylamide)
ESI-MS electrospray ionization-mass spectrometry
FSCE free solution capillary electrophoresis
GC gas chromatography
HEC hydroxyethylcellulose
HEMA hydroxyethyl methacrylate
HMPA hexamethyl phosphoric acid triamide
HPLC high performance liquid chromatography
HPMC hydroxypropylmethylcellulose
ID internal diameter
LC liquid chromatography
LPA linear polyacrylamide
MALDI-TOF-MS matrix-assisted laser desorption/ionization-time-of-flight mass
spectometry

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MEKC micellar electrokinetic chromatography
MPS ethacryloyloxypropyl trimethoxy silane
MS mass spectrometry
PB-PVS polybrene and poly(vinyl sulfonate)
PDMAA poly(dimethylacrylamide)
PEG poly(ethylene glycol)
PHEA Poly-N-hydroxyethylacrylamide
pI isoelectric point
PVA poly(vinyl alcohol)
RE-CZE reversed-charge Capillary Zone Electrophoresis
RSD relative standard deviation
SAM self-assembled monolayers
SDS sodium dodecyl sulfate
SDS-PAGE sodium dodecyl sulphate-polyacrylamide gel electrophoresis
TEGMA triethylene glycol monomethacrylate
Trp tryptophan
UV ultraviolet
VIS visible
WCID whole-column imaging detection













vi
List of Symbol

-1 -1κ molar extinction coefficient or absorptivity [M cm ]
-2η viscosity of the solution [Pa.s = N.s.cm ]
-3ρ density of the solution [kg.m ]
µ apparent electrophoretic mobility a
2 -1 -1µ electrophoretic mobility [m V s ] i
2 -1 -1µ EOF mobility [m V s ] eof
2 -1 -1µ apparent EOF mobility [m V s ] EOF app
ε dielectric constant
ζ zeta potential
-1v ion migration velocity [m.s ]
-1v net velocity of component i [m.s ] i(net)
-1v electroosmotic flow velocity [m.s ] eof
0 -1v migration velocity of component i [m.s ] i
z charge number of component i i
-19e elemental charge [1.602.10 C] 0
A peak area [AU.min]
c concentration of component i [M] i
d optical path length of light through the detection cell [cm]
-1E electric field strength [Vm ]
E( λ) adsorbed light intensity
F electric force [N] e
F drag forces [N] d
-1g gravitational acceleration (9.80665 N.kg )
Δh height difference [m]
I electric current [A]
I initial light intensity 0
I light intensity after absorbance t
L capillary length to detector or effective capillary length [cm] D
L distance between the electrodes or total capillary length [cm] T
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P injection pressure [mbar] inj
Δp pressure difference [Pa]
P additional pressure [mbar] add
q charge on the ion [C]
Q amount of species i i
r inner radius of capillary [m]
r ion radius i
-1R electric resistance of the electrolyte [V.A = Ω]
t migration time of component i [s] i
t mie of EOF marker [s] eof
t injection time [s] inj
t migration time [min] mig
U voltage [V]
3V injection volume [m ] i
















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