
Robert D.
Tilton
Center for
Complex Fluids Engineering, Department of Biomedical Engineering and Department
of Chemical Engineering, Carnegie Mellon University, Pittsburgh, PA 15213. tilton@andrew.cmu.edu
Microfluidic
Biotechnologies
Proteins and Protein-Conjugates at
Interfaces
Polymer and Surfactant
Co-Adsorption
Nanomaterials for
Environmental Remediation
The
main theme underlying the research in our group is the pervasive influence of
complex fluid interfacial phenomena in nature and technology. The group
maintains a long-standing interest in the structure, dynamics and function of
macromolecules or supramolecular assemblies at interfaces. In addition, a variety of concepts based on
colloidal and interfacial phenomena are being developed for new
biotechnological and environmental technologies. A selection of projects in the
group is presented below. A breakdown of publications in different areas is
included at the end.
Microfluidic Biotechnologies –
Electric Field-Driven Cell Separations
We are developing techniques based on a coupling of dielectrophoresis (DEP)
and electrohydrodynamic flows to manipulate colloidal particles and biological
cells using micropatterned electrodes.
Particles and cells migrate to either high field or low field traps,
according to their induced dipole strengths.
This offers the opportunity to separate mixtures according to
differences in dielectric properties.
Whereas DEP separations are typically operated at MHz frequencies, we
operate at frequencies ranging from ~ 10 Hz to 1 kHz where AC electroosmotic
flows are established along the edges of the microelectrode features. The simultaneous action of DEP and AC
electroosmotic drag forces on particles provides a finer level of
discrimination to laterally separate particles, making it possible to separate
mixtures that are not well separated by DEP (high frequency operation) methods
alone. The effect is easily visualized for the separation of low dielectric colloids from yeast cells in
suspension.

Lateral separation of polystyrene colloids and yeast cells. Low frequency (60 Hz) operation shown at left
aligns yeast cells along the microelectrode strip centers, with polystyrene
aligned along the strip edges. This placement is dictated by the DEP/AC
electroosmotic coupling. Operation at
high frequencies (1 MHz) shown at right instead aligns yeast in the high field
traps along the edges and polystyrene in the low field traps in the strip centers
and the centers of the insulating glass regions between strips. This placement is dictated by DEP alone. We have used this technique to separate
mixtures of suspension cells and to pattern vesicles. This work is an ongoing collaboration with Professor
Lee White. See Zhou et al. J. Colloid Interface
Sci. 285, 179-191 (2005). From
the Ph.D. Research of Hao Zhou.
Proteins at Interfaces – Effect of
Poly(Ethylene Glycol) Modification on Protein Adsorption
Covalently grafting PEG chains to protein therapeutics has been
demonstrated in the literature to improve pharmaceutical efficacy by
significantly increasing protein clearance times in the body. In collaboration with Professor Todd
Przybycien, we are investigating the effects of PEG grafts on the extent,
kinetics and reversibility of protein adsorption to solid surfaces, including
chromatographic materials such as will ultimately be used in the commercial
manufacture of PEGylated protein therapeutics and biodegradable
poly(lactide-co-glycolide) (PLG) materials that are used for sustained release
of proteins. Whereas the
protein-repellant nature of PEG has led to its perception as a biologically
“stealthy” polymer, PEG is surface active on many types of surfaces. From a
fundamental perspective, this makes PEG-protein conjugates an intriguing
adsorption system in that it can be considered as a complex block copolymer
with multiple surface active blocks.
Under some conditions, the conjugate can be adsorbed to a surface via
the protein “block” while under other conditions, the PEG block(s) anchor the
conjugate. We have found that on PLG
surfaces, PEGylation decreases the extent of ribonuclease A (a protein with
anti-tumoral effect) adsorption relative to the unmodified protein via an
excluded area effect on the surface.
PEGylation also reverses the way in which the protein responds to aging
of the PLG material. The diminished adsorption correlates with more complete
release, and improved retention of biological activity of ribonuclease A, from
PLG microspheres. See Daly et al.
“Adsorption of poly(ethylene glycol)-modified
ribonuclease A to a poly(lactide-co-glycolide) surface,” Biotechnology
and Bioengineering (2005)
On silica surfaces, steric interactions among PEG grafts on lysozyme
distort the shape of the adsorption isotherm and flip the preferred orientation
of the protein relative to the surface.
PEG grafts also increase the reversibility of adsorption (even though
PEG homopolymer adsorption and unconjugated lysozyme adsorption are both
effectively irreversible on silica). They inhibit the lateral aggregation of
lysozyme, thereby reducing a prevalent mode of surface-induced denaturation of
protein therapeutics. The complexity of
protein adsorption phenomena mandates the use of multiple experimental probes
in parallel. The conclusions described here were based on several novel
variants of total internal reflection fluorescence (TIRF) spectroscopy,
flouorescence spectroscopy, optical reflectometry, AFM colloidal force
measurements, electrokinetic (streaming current) measurements and attenuated
total reflection-Fourier transform infrared spectroscopy (ATR-FTIR). See Daly
et al. “Adsorption of poly(ethylene glycol) modified lysozyme to silica,” Langmuir 21, 1328-1337 (2005).

Adsorption isotherms on silica for lysozyme (squares), monoPEG-lysozyme
(triangles) and diPEG-lysozyme (circles).
From the Ph.D. research of Susan Daly
DiPasquo
Co-Adsorption of Polymers and
Surfactants – Control of Persistent Non-Equilibrium States in Thin Films
The properties of colloidal materials are engineered by controlling
interfacial chemistry. Mixed
self-assembly and/or co-adsorption of polymers and surfactants exert powerful
controls over the macroscopic properties of complex fluids that are found in
the manufacture and application of materials such as pharmaceutical suspensions
or solid dosage forms, ceramics, paints, inks, and personal care products. The
tendency of surfactants and polymers to exert strongly coupled influences on
complex fluid properties makes these fluids particularly difficult to
formulate. A particular emphasis of our
research in this area concerns the prevalence of persistent non-equilibrium, or
hysteretic, states in polymer layers. We
have found that it is possible to control the composition and structure of
polymer layers by controlling the order in which surfactants and polymers are
exposed to a surface. For example, dense films of Pluronic® PEO-PPO-PEO
copolymers can be prepared on silica by co-adsorption with sodium dodecyl
sulfate (SDS), followed by removal of the surfactant from the system. Similar SDS-processing can be used to to
condense, or to stretch, cellulosic polymers on a silica surface depending on
conditions.


The force between two silica surfaces coated with PEO-PPO-PEO
layers, measured by atomic force microscopy, on the left is for a layer
prepared by simple adsorption from an SDS-free solution. The force profile on the right, which is
consistent with a thinner layer, is for a PEO-PPO-PEO layer produced by co-adsorption
with SDS followed by surfactant removal.
Although the SDS-processed layer is thinner, it contains approximately
30% more polymer per unit area – the processed layer is ~ 50 % denser, due to a
condensation of polymer around bound SDS micelles that occurs during
adsorption. Once deposited on the
surface, the condensed polymer configuration does not relax sufficiently to
match the configuration of the “ordinary” polymer layer. See Braem et al. “Control of Persistent
Nonequilibrium Adsorbed Polymer Layer Structure by Transient Exposure to
Surfactants,” Langmuir 19, 2736 (2003). From
the Ph.D. Research of Alan Braem and Derek Berglund.
Nanomaterials for
Environmental Remediation – Development of Amphiphilic Polymers for Targeted
Delivery of Iron Nanoparticles to DNAPL/Water Interfaces
Organic contamination of subsurface soil and groundwater is an extensive
and vexing environmental problem that stands to benefit from nanotechnology.
The Environmental Protection Agency reports that contamination by organic
pollutants, especially chlorinated volatile organic compounds (CVOCs), are
primary concerns at over half of the Superfund National Priorities List sites.
(See Common Chemicals Found at Superfund
Sites, last update
In collaboration with Professors Gregory Lowry (Dept.
of Civil & Environmental Engineering), Krzysztof Matyjaszewski (Dept. of
Chemistry) and Sara Majetich (Dept. of Physics), we are developing the concept
of targeted delivery of remediation agents to the contamination source –
underground residuals of dense nonaqueous phase liquids (DNAPL) trapped in
microporous aquifers. We have synthesized
amphiphilic poly(methacrylic
acid)-b-poly(methylmethacrylate)-b-poly(styrenesulfonate) triblock copolymers
that adsorb to reactive nanoparticulate Fe(0) (RNIP, particles commercially
available from Toda

The macroscale problem.
Illustration of DNAPL distribution as residual saturation and a plume of
dissolved contaminants in an aquifer. Pump and treat remediation methods
require multiple wells to pump groundwater from the plume to the surface for
chemical treatment, leaving the residual saturation source behind to
continually replenish the plume. Our
proposed method will target the chemical treatment to the residual saturation
zone in situ. Drawing from S.G. Huling and J.W. Weaver, “Dense Nonaqueous Phase Liquids,” EPA Groundwater
Issue EPA/540/4-91-002, 1991.

The nanoscale solution. Residual DNAPL (shown in
gray) is trapped in and between soil grains (shown in black) in pores that
range from micrometers to as small as 2 – 50 nm in diameter. Targeted iron nanoparticles, by virtue of
their size and surface functionalization, gain access to the trapped DNAPL in
micropores and adsorb with high affinity to the DNAPL/water interface. There, the accumulated iron can reduce DNAPL
to nontoxic products.

The targeting mechanism.
Particle-grafted block copolymers contain a hydrophilic (blue) block
that has an affinity for water and a hydrophobic (red) block that has an
affinity for DNAPL. In water, the hydrophilic block swells and the hydrophobic
block collapses. The reverse happens in
the DNAPL phase. This amphiphilicity anchors the particle at the DNAPL/water
interface. The hydrophilic block is
large enough to stably suspend particles in water without aggregating.
Incorporating strong negative charge in the hydrophilic block minimizes
particle adhesion to negatively charged mineral or natural organic matter
surfaces in the soil before reaching the DNAPL.
Grafted Nanoparticles

As a sub-set of this
project, we developed poly(styrenesulfonate)-grafted silica nanoparticles that
are excellent stabilizers of non-invertable oil-in-water emulsions. These “
From the Ph.D. research of Kevin Sirk and Navid Saleh.
YKI Ambassador Program
Professor Tilton maintains an active collaboration with the Institute
for Surface Chemistry (Ytkemiska Institutet; YKI) in
As a YKI Ambassador, Prof. Tilton is
available to facilitate industrial research collaborations with YKI, including
joint collaborations that leverage the knowledge and resources of YKI and Prof.
Tilton’s Complex Fluid and Bio-Interfaces Group at Carnegie Mellon.
Group Publications
Polymers and Surfactants: Adsorption,
Self-Assembly, Wetting, Surface Forces
K. D. Berglund, M. T. Truong, R. D. Tilton and L. M. Walker, “Rheology of Transient Networks Containing Hydrophobically Modified Cellulose, Anionic Surfactant and Colloidal Silica: Role of Selective Adsorption,” Rheol. Acta 43, 50 – 61 (2004)
O.J. Rojas, P.M. Claesson, K.D. Berglund, and R.D. Tilton, “Coadsorption and Surface Forces for Selective Surfaces in Contact with Aqueous Mixtures of Oppositely Charged Surfactants and Low Charge Density Polyelectrolytes,” Langmuir, 20, 3221-3230 (2004).
I.N.
Nnebe, R.D. Tilton, and J.W. Schneider, “Direct Measurement of the Stability of
Poly(ethylene glycol)-Poly(ethylenimine) Graft Films,” Journal of Colloid and Interface Science 276, 306-316 (2004).
R.D. Tilton, “Adsorption from Mixtures of Polymers and Surfactants,” in Encyclopedia of Surface and Colloid Science, 8th Updated Edition, P. Somasundaran, Ed., Marcel-Dekker, Inc., New York (2004)
K.D. Berglund, A. Timko, T.M. Przybycien, R.D. Tilton, “Use of Nonionic Ethylene Oxide Surfactants as Phase Transfer Catalysts for Poly(Acrylic Acid) Adsorption to Silica against an Electrostatic Repulsion,” Progress in Colloid and Polymer Science 122, 56-66 (2003).
K.D. Berglund, T.M. Przybycien, R.D. Tilton, “Coadsorption of Sodium Dodecylsulfate with Hydrophobically Modified Nonionic Cellulose Polymers I: Role of Polymer Hydrophobic Modification,” Langmuir 19, 2705-2713 (2003).
K.D. Berglund, T.M. Przybycien, R.D. Tilton, “Coadsorption of Sodium Dodecylsulfate with Hydrophobically Modified Nonionic Cellulose Polymers II: Role of Surface Selectivity in Adsorption Hysteresis,” Langmuir 19, 2714-2721 (2003).
A.D. Braem, S. Biggs, D.C. Prieve, R.D. Tilton, Control of Persistent Nonequilibrium Adsorbed Polymer Layer Structure by Transient Exposure to Surfactants,” Langmuir 19, 2736 (2003).
N. Kumar, K. Varanasi, R.D. Tilton, S. Garoff, “Surfactant Self-Assembly ahead of the Contact Line on a Hydrophobic Surface and its Implications for Wetting,” Langmuir 19, 5366 (2003).
S.B. Velegol, R.D. Tilton, “Specific Counterion Effects on the Competitive Co-Adsorption of Polyelectrolytes and Ionic Surfactants,” Journal of Colloid and Interface Science 249, 282-289 (2002).
E.A.S. Doherty, K.D. Berglund, B.A. Buchholz, I.V. Kourkine, T.M. Przybycien, R.D. Tilton, A.E. Barron, “Critical Factors for High-Performance Physically Adsorbed (Dynamic) Polymeric Wall Coatings for Microchannel Electrophoresis of DNA,” Electrophoresis 23, 2766-2776 (2002).
S.B. Velegol, R.D. Tilton, “A Connection between Interfacial Self-Assembly and the Inhibition of Hexadecyltrimethylammonium Bromide Adsorption on Silica by Poly-L-lysine,” Langmuir 17, 219 (2001).
A.D.
B.B. Luokkala, S. Garoff, R.D. Tilton, R.M. Suter, “Interfacial Structure and Rearrangement of Nonionic Surfactants near a Moving Contact Line,” Langmuir 17, 5917 (2001).
S.B. Velegol, B.D. Fleming, S. Biggs, E.J. Wanless, R.D. Tilton, “Counterion Effects on Hexadecyltrimethylammonium Surfactant Adsorption and Self-Assembly on Silica,” Langmuir 16, 2548 (2000).
J.-H. Kim, M.M. Domach, R.D. Tilton, “Effect of Electrolytes on the Pyrene Solubilization Capacity of Dodecyl Sulfate Micelles,” Langmuir 16, 10037 (2000).
J.-H. Kim, M.M. Domach and R.D. Tilton,
“Solubilization Capacity of Pyrene in Octaethylene Glycol Monododecyl Ether
Micelles,” Colloids Surf. A Physicochem.
J.-H. Kim, M.M. Domach, R.D. Tilton, “Pyrene Micropartitioning and Solubilization by Sodium Dodecyl Sulfate Complexes with Poly(ethylene glycol),” J. Phys. Chem. B, 103, 10582 (1999).
R.D. Tilton, “Scanning Angle Reflectometry and its Application to Polymer Adsorption and Coadsorption with Surfactants,” in Polymer-Colloid Interactions: Techniques and Applications, P. Dubin and R. Farinato, Eds., pp. 331-363, John Wiley and Sons, Inc., New York (1999).
E.S.
E.S. Pagac, R.D. Tilton and D.C. Prieve, “Depletion Attraction Caused by Unadsorbed Polyelectrolytes,” Langmuir 14, 5106 (1998).
E.S. Pagac, D.C. Prieve, Y. Solomentsev and R.D. Tilton, “A Comparison of Polystyrene-Poly(ethylene oxide) Diblock Copolymer and Poly(ethylene oxide) Homopolymer Adsorption from Aqueous Solutions,” Langmuir, 13, 2993-3001 (1997).
J.R. Charron and R.D. Tilton, “Penetration of Insoluble Lipid Monolayers at the Air-Water Interface by Water-Soluble Block Copolymers and Homopolymers,” Langmuir 13, 5524 (1997).
J.R. Charron and R.D. Tilton, “A Scanning Angle Reflectometry Investigation of Block Copolymer Adsorption to Insoluble Lipid Monolayers at the Air-Water Interface,” J. Phys. Chem. 100, 3179 (1996).
E.M. Furst, E.S. Pagac, and R.D. Tilton,
“Coadsorption of Polylysine and the Cationic Surfactant Cetyltrimethylammonium
Bromide on Silica,” Ind.
E.S. Pagac, R.D. Tilton and D.C. Prieve,
“Hindered Mobility of a Rigid Sphere Near a Wall,” Chem.
Colloidal Dispersions
N. Saleh, T. Sarbu, K. Sirk, G.V. Lowry, K. Matyjaszewski, and R.D. Tilton, “Oil-in-water emulsions stabilized by highly charged polyelectrolyte-grafted silica nanoparticles,” Langmuir in press (available on-line July 2005).
Proteins at Interfaces
S.M. Daly, T.M. Przybycien and R.D. Tilton, “Adsorption of poly(ethylene glycol)-modified ribonuclease A to a poly(lactide-co-glycolide) surface,” Biotechnology and Bioengineering 90, 856-868 (2005).
S. Daly, T.M. Przybycien, and R.D. Tilton, “Adsorption of poly(ethylene glycol) modified lysozyme to silica,” Langmuir 21, 1328-1337 (2005).
S. Daly, T.M. Przybycien, R.D. Tilton, “Coverage Dependent Orientation of Lysozyme Adsorbed on Silica,” Langmuir 19, 3848 (2003).
E. J. Choi,
M.D. Foster, S. Daly, R. Tilton, T. Przybycien, C.F. Majkrzak, P. Witte, H.
Menzel, "Effect of Flow on Human Serum Albumin Adsorption to
Self-Assembled Monolayers of Varying Packing Density, " Langmuir 19, 5464 (2003).
R.D. Tilton, “Mobility of Biomolecules at Interfaces,” in Biopolymers at Interfaces, Surfactant Science Series vol. 75, M. Malmsten, Ed., pp. 363-407, Marcel Dekker, New York (1998). Reproduced in Biopolymers at Interfaces, 2nd Edition Revised and Expanded, Surfactant Science Series vol. 110, M. Malmsten, Ed., pp. 221-257, Marcel Dekker, New York (2003).
M.L. Sun, R.D. Tilton “Coadsorption of Protein/Surfactant Complexes to the Air/Water Interface,” Colloids and Surfaces B: Biointerfaces 20, 281 (2001).
S. M. Butler, M. A. Tracy and R.D. Tilton, “Adsorption of Serum Albumin to Thin Films of Poly(lactide-co-glycolide),” J. Controlled Release 58, 335 (1999).
E.F. Tsung and R.D. Tilton, “Measurement of Catalytic Reaction Kinetics for Adsorbed Enzyme Monolayers,” J. Colloid Interface Sci. 213, 208 (1999).
J.L. Robeson and R.D. Tilton, “Spontaneous Reconfiguration of Adsorbed Lysozyme Layers Observed by Total Internal Reflection Fluorescence with a pH-Sensitive Fluorophore,” Langmuir, 12, 6104 (1996).
J.L. Robeson and R.D. Tilton, “Effect of Concentration Quenching on Fluorescence Recovery After Photobleaching Measurements,” Biophys. J. 68, 2145 (1995).
E. Blomberg, P.M. Claesson and R.D. Tilton, “Short-Range Interaction between Adsorbed Layers of Human Serum Albumin,” J. Colloid Interface Sci 166, 427 (1994).
T.E. Rabe and R.D. Tilton, “Surface Diffusion of Adsorbed Proteins in the Vicinity of the Substrate Glass Transition Temperature,” J. Colloid Interface Sci. 159, 243 (1993).
R.D. Tilton,
R.D. Tilton, C.R. Robertson and A.P. Gast, “Lateral Diffusion of Bovine Serum Albumin Adsorbed at the Solid-Liquid Interface,” J. Colloid Interface Sci. 137, 192 (1990).
R.D. Tilton, A.P. Gast and C.R. Robertson, “Surface Diffusion of Interacting Proteins: Effect of Concentration on the Lateral Mobility of Adsorbed Bovine Serum Albumin,” Biophys. J. 58, 1321 (1990).
Dielectrophoresis and
Electrohydrodynamics
H. Zhou, M.A. Preston, R.D. Tilton, and
L.R. White, “Calculation of the dynamic impedance of the double layer on a
planar electrode by the theory of electrokinetics,” Journal of Colloid and Interface Science in press (available on-line
H. Zhou, L.R. White and R.D. Tilton, “Lateral separation of colloids or cells by dielectrophoresis augmented by AC electroosmosis,” Journal of Colloid and Interface Science 285, 179-191 (2005).
H. Zhou, M.A. Preston, R.D. Tilton, and L.R. White, “Calculation of the electric polarizability of a charged spherical dielectric particle by the theory of colloidal electrokinetics,” Journal of Colloid and Interface Science 285, 845-856 (2004).
V. Brisson, R.D. Tilton, “Self-assembly and two-dimensional patterning of cell arrays by electrophoretic deposition,” Biotechnology and Bioengineering 77, 290-295 (2002)
Environmental Applications
Y.Liu, S. Majetich, K. Matyjaszewski, R.D. Tilton, D. Sholl, and G.V. Lowry, “TCE Dechlorination Rates, Pathways, and Efficiency of Nanoscale Iron Particles with Different Properties,” Environmental Science and Technology, 39, 1338-1335 (2005).