Chapter 17

Protein—Surfactant Interactions at Solid

Surfaces

Thomas Arnebrant and Marie C. Wahlgren

Department of Food Technology, University of Lund, P.O. Box 124,

221 00 Lund, Sweden
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Effects of surfactants on protein adsorption are reviewed. Differences

between removal of preadsorbed proteins (elutability) and competitive
adsorption are discussed and simple models are suggested. It can be
concluded that surfactants may interact through solubilization or
replacement mechanisms depending on surfactant- surface interactions
and surfactant- protein binding. Solubilization requires complex
formation between protein and surfactant, and the replacement
adsorption of the surfactant to the surface. As for protein adsorption,
one of the most important properties affecting the elutability appears to
be the conformational stability. Differences between a competitive
situation and addition of surfactant after adsorption of the protein are
suggested to originate from alteration in surface activity of protein­
-surfactant complexes formed in solution as compared to pure protein,
the difference in diffusivity of surfactants and protein, and time
dependent conformational changes of the protein.

Interactions between surfactants and proteins take place in various applications

involving proteins in contact with solid surfaces. Examples are found in general
detergency, for example when process equipment should be cleaned from deposits of
protein origin. Surfactants and proteins or peptides may be simultaneously present
during protein isolation procedures and for minimizing loss of active substance during
drug administration. Another area of application of these interactions is in the field of
dentistry where so-called anti-plaque agents are used in the treatment of plaque
related diseases (1-4). The degree of removal of adsorbed protein by surfactants has
also been used as an indication of the mode of protein attachment to the surface, in
particular with respect to time dependent conformational changes (5-7). The general
features of protein adsorption involve what is referred to as multiple states of ad­
sorption cf. (8). This means that adsorbed protein may exist in several adsorbed frac­
tions with varying binding modes to the surface. These different fractions may be
distinguished by their differences in binding strength as indicated by the degree of
removal by rinsing with buffer, addition of surfactant or by their different suscep­
tibility to exchange by the same or other types of protein. The importance of
structural features of interfacial layers of protein is recognised in research focused on
e.g. biocompatible materials, dental pellicle buildup, immuno assays and enzyme

0097-6156/95/0602-0239$12.00/0
© 1995 American Chemical Society

In Proteins at Interfaces II; Horbett, T., et al.;

ACS Symposium Series; American Chemical Society: Washington, DC, 1995.
 240 PROTEINS AT INTERFACES II

immobilisation. A correlation between the above mentioned structural aspects and a

simple measurable quantity as for example surfactant mediated elutability is therefore
convenient from a practical point of view, and has proved to be valuable in the
assessment of the performance of biomedical polymers (9,70).

Surfactant Adsorption at Solid Surfaces

Surfactants, as the name implies, tend to adsorb at most interfaces and thereby
strongly reduce the interfacial free energy. Surfactants are usually classified accord­
ing to their head group as an-, cat- and nonionic, respectively. The surfactants
discussed in this paper are presented in table I. The properties of the hydrophobic,
usually hydrocarbon, part as well as the hydrophilic head group, will affect the affin­
ity of the molecule for interfaces. Furthermore, due to the strong tendency for the
hydrophobic chains to avoid contact with water, self-association will take place both
in solution and at interfaces, a fact that will influence the adsorption behaviour. In
bulk phase, self-association usually involves the formation of spherical micelles at
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low concentration and depending on surfactant structure, cylindrical micelles,

lamellar structures, cubic phases and structures of the reversed type may form at
higher concentrations and appropriate conditions (77). The effect is that the monomer
concentration in solution will be strongly dependent on the association pattern (//)
and thus have a pronounced effect on the interfacial behaviour.

Table I : A presentation of the surfactants discussed in the text

Surfactant Abbreviation cmc Headgroup
Sodium dodecylsulphate SDS 8mM -SOr
Dodecyltrimethylammonium DTAB 15 mM -N+-(CH )3 3

bromide
Tetradecyltrimethylammonium TTAB 3.6 mM -N+-(CH ) 3 3

bromide
Cetyltrimethylammonium CTAB 0.9 mM -N+-(CH )3 3

bromide
Triethylene glycol C12E3 - -(OCH CH ) OH
2 2 3

monododecyl ether
Pentaethylene glycol C12E5 0.065 mM - ( O C H C H ) O H
2 2 5

monododecyl ether
Octaethylene glycol Ci E 2 8
0.071 m M -(OCH CH )80H 2 2

monododecyl ether

To state briefly, a few general features of surfactant adsorption are the

following:

1) At high surfactant concentrations (around the cmc) as discussed in the present

work, the adsorption rate is fast, as exemplified in Fig. 1 for the adsorption of
D T A B to methylated silica.
2) For water soluble surfactants, the adsorption is reversible upon dilution which
also is illustrated in Fig. 1.
3) Usually a plateau in the adsorption isotherm is reached in the range of the cmc
(critical micelle concentration), Fig. 1.
4) As a rule surfactants adsorb at hydrophobic surfaces. The amounts adsorbed are
in the range of, or below, those corresponding to a monolayer, Fig. 2.

In Proteins at Interfaces II; Horbett, T., et al.;

ACS Symposium Series; American Chemical Society: Washington, DC, 1995.
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ACS Symposium Series; American Chemical Society: Washington, DC, 1995.
In Proteins at Interfaces II; Horbett, T., et al.;

Fig. 1 The adsorption of D T A B ( 2*cmc in phosphate buffered saline pH

7, 1=0.17) onto methylated silica. Data are from (25) (left) A schematic
illustration of an isotherm for adsorption of ionic surfactants to hydrophilic
surfaces, from Somasundaran (76)(right).
 242 PROTEINS AT INTERFACES II

5) In the absence of specific chemical interactions, ionic surfactants only adsorb

onto hydrophilic surfaces of opposite charge(72-76). At these surfaces bilayers or
corresponding structures are formed (see below), Fig. 2.
6) Isotherms for surfactants adsorbing to hydrophilic surfaces usually have the
features shown in Fig. 1 (76). The different regions will depend on the asso­
ciation of the surfactant molecules at the interface and in solution. There is a vast
literature concerning the association of surfactants at solid/water interfaces (72-
23). Fig. 2 is an illustration of possible association behaviour of surfactants in the
different regions.

The structure of the surface aggregates at the plateau has been debated and
surface micelles, finite bilayers or infinite bilayers have been suggested for
hydrophilic surfaces. Indications of complete bilayers (24) (Fig. 2 iv a) or
interpenetrating hydrocarbon chains (76) (Fig. 2 iv b) have been found.
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Surfactant-Protein Interactions in Solution

Publication Date: May 5, 1995 | doi: 10.1021/bk-1995-0602.ch017

For mixtures of proteins and surfactants there might be an interaction in bulk solution
involving the formation of surfactant-protein complexes which have different
properties from those of the pure protein (26,27). Further, the binding of surfactant to
protein will reduce the concentration of free surfactant molecules available for inter­
action with the protein at the interface which may show up as an apparent increase in
the cmc.
Ionic surfactants are known to interact with proteins in solution, and the inter­
action is generally stronger for SDS than for cationic surfactants (26-31). Nonionic
surfactants are known to generally interact poorly with soluble proteins (26). Three
types of interactions are observed (26):

i) Binding of surfactant by electrostatic or hydrophobic interactions to specific

sites in the protein, such as for |3-lactoglobulin (26, 32, 33) and serum albumin
(26,28,34).
ii) Cooperative adsorption of surfactant to the protein without gross conformational
changes.
iii) Cooperative binding to the protein followed by conformational changes (28-30,
35,36)

The changes i) -iii) can occur in the same system when surfactant concentration
is increased. The conformational changes that occur in case (iii) involves changes in
secondary structure (28, 29, 35). It is assumed that the surfactant molecules bind to
the polypeptide chain and several models for the protein surfactant complexes have
been suggested e.g., rigid rod (37), pearl and necklace (38) and flexible helix model
(39). In the cooperative region (ii-iii), above the critical association concen­
tration (cac), the interaction is mainly of hydrophobic character (26,29,36).

Surfactants and Proteins at Interfaces

Interaction of Surfactants with Adsorbed Proteins. The removal of preadsorbed

proteins by surfactant has been extensively studied by Horbett and co-workers (5, 6,
40, 41) in investigations into the adsorption strength of proteins, particularly
fibrinogen. They introduced the term "elutability" in order to describe the degree of
removal. The degree of surfactant elutability of proteins is affected by factors that are
known to influence the binding strength of a protein to a surface. Thus, surfactant
elutability has been found to decrease with factors favouring conformational changes

In Proteins at Interfaces II; Horbett, T., et al.;

ACS Symposium Series; American Chemical Society: Washington, DC, 1995.
 17. ARNEBRANT & WAHLGREN Protein-Surfactant Interactions 243

i. e. decreasing protein concentration^, 6,40\ increasing temperature (5, 6), time of

adsorption "residence time", (5, 6, 40, 41) and decreasing stability of the protein (42-
44). However, surfactant elutability will not only be influenced by protein properties
but also by the type of surfactant (6, 45-47) and surface (7, 40, 44-46, 48, 49) as
discussed in more detail below. Of course, the self association of the surfactant plays
a major role in this context and the use of non associating displacers may be more
straightforward if evaluation of the binding strength is the main concern.
A discussion on the influence on protein-surfactant behaviour by protein proper­
ties, surfactant properties and surface properties, based on our own observations as
well as by other workers in the field is given below.

Influence of Protein Properties. As discussed above, the interaction between

surfactant and protein might involve a certain specificity, especially so at low surfac­
tant concentrations. At higher surfactant concentrations these effects are less pro­
nounced. The correlation of protein properties to elutability is not straightforward, as
the effects of different properties overlap. However, when the D T A B induced elution
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Publication Date: May 5, 1995 | doi: 10.1021/bk-1995-0602.ch017

of six model proteins, cytochrome c, bovine serum albumin, a-lactalbumin, (J-

lactoglobulin, lysozyme and ovalbumin adsorbed at a silica surface was compared,
the removal of the proteins that were still adsorbed after rinsing with buffer, appear to
increase with decreasing molecular weight and adiabatic compressibility (a measure
of conformational stability (50)) and increasing thermal denaturation temperature
(44). In the case of a methylated silica surface, the trends were weaker and differences
between the proteins as regards elutability were smaller. However, increasing
molecular weight and shell hydrophobicity of the protein seem to reduce elutability. It
was also found that the elutability did not relate to the degree of desorption of
proteins upon rinsing with buffer, indicating that the two mechanisms are different.
Recent experiments on stability mutants of bacteriophage T4 lysozyme show a very
convincing relation between D T A B mediated elutability and the difference in free
energy of thermal unfolding of the protein in comparison with the wild type (see sep­
arate contribution within this volume (43)). It might thus be concluded that factors
relating to the structural stability of the protein is of major importance and that an
increased stability increases the degree of elution.

Influence of Surfactant Properties. It is necessary to keep in mind that the

surfactants will, depending on mechanism of elution (see below), interact with the
protein, the surface or usually both. Therefore knowledge of the main mechanism of
removal for each combination of surfactant, protein and surface is mandatory in order
to correctly interpret the effect of one component. The influence of different surfac­
tant headgroups on the desorption of lysozyme at hydrophilic silica surfaces is pre­
sented in Fig. 3 (25,44,45).
Surfactant concentrations differ in this figure but are in all cases above the cmc,
except for C 1 2 E 3 which does not form micelles (57). The difference between SDS,
cationic surfactants and nonionics as regards the effect on surfactant elutability of
proteins is analogous to the strength of binding to protein in solution. This suggests
that above the critical association concentration (cac), complex formation between
surfactant and protein is involved in the removal mechanism on hydrophilic surfaces.
In this connection, Blomberg and coworkers studied the removal of adsorbed
lysozyme by SDSo (Sodium Dodecane Sulfonate) and SDS. They found that SDSo,
which has a Krafft temperature above room temperature and hence does not form
micelles, had a very minor effect on the interaction between adsorbed lysozyme
layers on mica (52) and concluded that few surfactant molecules were bound to the
adsorbed protein. SDS showed a similar low binding to lysozyme on mica at low con­
centrations (up to 0.5 cmc) but caused a collective desorption of the protein at the cmc
of the surfactant, indicating that the cac to adsorbed lysozyme is in the range of its

In Proteins at Interfaces II; Horbett, T., et al.;

ACS Symposium Series; American Chemical Society: Washington, DC, 1995.
 244 PROTEINS AT INTERFACES II

M M M t 'M^MWM JI111I<
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Fig. 2. An illustration of probable arrangements of adsorbed surfactant

molecules at different degrees of surface coverage. Adsorption to
hydrophilic surfaces (left panels) and hydrophobic ones (right panels). The
illustrations are drawn to represent structures having minimal water contact
with the hydrophobic parts of the molecules. The figures should be
considered as schematic and other structures, especially for ii-iii, have been
suggested (72, 76,22,23,25).

0.30

^0.20

Silica

S0.10
T3
<

0.00
3600 5400 7200
Time (s)
Fig. 3. The elutability of lysozyme by different surfactants. The adsorbed
amount versus time for adsorption of lysozyme to silica followed by buffer
rinsing after 1800 sec, addition of surfactant after 3600 seconds and a final
rinse with buffer after 5400 sec. The protein concentration is 1 mg/ml in
phosphate buffered saline solution pH 7,1=0.17. The surfactants are 5 mg/ml
SDS (•), 5 mg/ml D T A B (O), 0.5 g/ml C12E3 (•), 0.5 mg/ml C12E5 (+).

In Proteins at Interfaces II; Horbett, T., et al.;

ACS Symposium Series; American Chemical Society: Washington, DC, 1995.
 17. ARNEBRANT & WAHLGREN Protein-Surfactant Interactions 245

self-association limit in solution (cmc) (53) and further supports that the formation of
surfactant- protein complexes are important in the removal process.
The non-ionic surfactants do not remove lysozyme from the hydrophilic silica
surface. As mentioned above, these surfactants bind to a very low extent to protein in
solution and to the protein covered surface (fig. 3).
It was found that removal of protein at methylated silica surfaces (hydrophobic)
is similar for the different surfactants (25), also for non-micelle forming ones, indi­
cating that the proteins are removed through replacement due to higher surface
activity of the surfactant. The trend, that non-ionic surfactants do not affect the
amount adsorbed at hydrophilic surfaces but have a considerable effect at
hydrophobic surfaces has also been observed by Elwing et al. (54), when studying
surfactant elutability of proteins adsorbed at a surface with a gradient in wettability.
Rapoza and Horbett (6) found that surfactants with large headgroups such as Tween-
20 gave rise to lower fibrinogen elutability levels than other surfactants at
polyethylene surfaces. Welin-Klintstrom et al. (47) found that the elutability of
fibrinogen adsorbed at wettability gradient surfaces decreased with the bulkiness of
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the hydrophobic part of the surfactant. In this connection it was also found that non-
ionics showed an increased removal of fibrinogen into the more hydrophilic region of
the gradient surface when the cloud point (phase separation temperature) was
approached (48). Further, these general observations of removal efficiency are in line
with the findings of Backstrom and co-workers (48,55, 56) who studied the removal
of fat by different surfactants and found a maximum at conditions corresponding to an
optimum in the packing of surfactant molecules at a flat interface and those of
Malmsten and Lindman (57) who investigated, among other variables, the effect of
temperature on cleaning of hard surfaces.
The effects of chain length of alkyltrimethylammonium surfactants on the
elutability of fibrinogen are presented in Fig. 4 (25). The dependence of the elutability
of proteins on chain length of surfactant is small at both silica and methylated silica.
The elutability of proteins by DTAB is slightly smaller than for TTAB and C T A B .
Rapoza and Horbett (6, 58) did not find any effects of chain length of sodium
alkyl sulphates on the elutability for fibrinogen and albumin down to a chain length of
six methyl groups. However, they found, as expected, that the chain length did in­
fluence the surfactant concentration at which the onset of protein removal started. The
trend was similar to the one observed for the onset of cooperative binding events (e.g.
micelle formation).
It may be concluded that surfactant headgroup effects are most pronounced at
hydrophilic surfaces but less important at hydrophobic ones. In addition, it appears
that principles for detergency in general, involving the packing efficiency of
molecules at interfaces are qualitatively applicable in these systems.

Influence of Surface Properties. As described above, the adsorption and orientation

of surfactant are dependent on the type of surface and it is therefore natural to expect
that the way in which proteins are removed by surfactants should be influenced by the
surface character as well. Elwing et al. (49, 54) studied the surfactant elutability of
proteins adsorbed to a gradient in hydrophobicity and found large differences in the
amounts removed at the hydrophilic and hydrophobic ends. In the case of a non ionic
surfactant (Tween 20) the elutability was largest at the midpoint of the gradient (54),
which might be attributed to enhanced conformational changes of the adsorbed
protein at the hydrophobic end in combination with a lower efficiency of nonionics at
hydrophilic surfaces. Horbett and co-workers (5,40) studied the elutability of fibrino­
gen and albumin at different polymeric surfaces and found that the elutability and the
change of elutability with time differed among the surfaces. These differences could
not, however, be correlated to their critical surface tension of wetting.

In Proteins at Interfaces II; Horbett, T., et al.;

ACS Symposium Series; American Chemical Society: Washington, DC, 1995.
 246 PROTEINS AT INTERFACES II

0.8 r

1
w>0.6 -
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1 1 1
0.0 ' "
0 2000 4000 6000 8000
Time (s)

Fig. 4. The elutability of fibrinogen by different surfactants. The protein

concentration is 0.4 mg/ml in phosphate buffered saline solution pH 7. The
adsorption procedure is the same as in Fig. 3 and the surfactants are SDS (x),
D T A B (+), T T A B (O), C T A B (•) and surfactant concentration is twice the
cmc.

In Proteins at Interfaces II; Horbett, T., et al.;

ACS Symposium Series; American Chemical Society: Washington, DC, 1995.
 17. ARNEBRANT & WAHLGREN Protein-Surfactant Interactions 247

Investigations into the elutability of lysozyme and p-lactoglobulin on methy­

lated silica (hydrophobic) and oxides of silicon, chromium and nickel showed that the
elutability did not follow any clear cut rules relating to the charge on the surface.
Instead, elutability of P-lactoglobulin and lysozyme by SDS decreased in the order
silica>chromium oxidonickel oxide. The order of decrease is nearly the same for
C T A B , but the difference between nickel and chromium oxides was insignifi­
cant (45). The similarity between the two oppositely charged surfactants indicates that
the elutability in these cases mainly reflects the binding strength of the protein to the
surface.
At methylated silica surfaces the elutability was high. This should, however, not
be considered as evidence for weak binding of the proteins to the surface but rather as
an indication of the strong interaction between the surfactants and surface.
It might be suggested that, even though not of universal applicability, an electro­
static repulsion between surfactant and surface or a strong (hydrophobic) interaction
between the hydrocarbon chain of the surfactant and the surface might favour the
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elution of adsorbed protein.

Publication Date: May 5, 1995 | doi: 10.1021/bk-1995-0602.ch017

Simple Models and Conclusions. We have found it useful to classify the observed
effects of surfactants on adsorbed proteins in four categories. A short description of
these follows below (see Fig. 9 and Figs 5-8):
i) No remaining adsorbate after addition of surfactant. At these conditions, the
surfactant does not adsorb to the surface. However, it interacts with the protein
and forms a complex that desorbs from the surface Fig. 5
ii) Replacement of protein by surfactant. This implies that the interaction between
surfactant and surface has to be stronger than the interaction between protein or
surfactant/protein complex and surface. Adsorption of the surfactant to the
surface is essential in this connection, but binding of surfactant to protein may
not be required Fig. 6.
iii) The surfactant might adsorb to the surface and/or to adsorbed protein but does
not have any net effect on the amount of protein adsorbed Fig. 7.
iv) The adsorbate is only partly removed by the surfactant. Partial elutability of
proteins has previously been suggested as an indication of the presence of
multiple states of adsorbed proteins (8) Fig. 8.

It is important to note that surfactant can remove proteins without binding to the
surface. This could be described as a solubilization of the proteins by the surfactant.
An interesting question is whether the replacement of proteins by surfactant in
category (ii) is first initiated by solubilization followed by surfactant adsorption?
Non-ionic surfactants interact to a very low extent with soluble proteins (26), as for
example can be concluded from the low amounts adsorbed to the adsorbed protein
layer at the silica surface (Fig. 3). These surfactants still remove proteins from the
methylated silica surface, and it is thus evident that in this case it occurs through re­
placement of adsorbed protein molecules by surfactant.

Adsorption from Mixtures of Proteins and Surfactants

The adsorption from surfactant/protein mixtures to hydrophobic solid surfaces is to

some extent analogous to the adsorption at air/water or oil/water interfaces which
have been the subject of frequent studies (34, 59, 60) due to the importance of
stability of food emulsions and foams. Competitive adsorption between proteins and
surfactant at these interfaces has recently been reviewed by Dickinson and Woskett
(67). They conclude that, as expected, small surface active components above a
certain critical concentration will dominate over proteins at these interfaces, as such

In Proteins at Interfaces II; Horbett, T., et al.;

ACS Symposium Series; American Chemical Society: Washington, DC, 1995.
 248 PROTEINS AT INTERFACES II

r(ugfcrn )
0.3
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2000 4000 8000

Time (s)

Fig. 5. The adsorbed amount versus time for adsorption of lysozyme to

silica followed by buffer rinsing after 1800 sec, addition of surfactant (SDS)
after 3600 seconds and a final rinse with buffer after 5400 sec. ( O ) .
Adsorption from a mixture of the protein and surfactant for 1800 sec.
followed by rinsing is also included (•). The experiments were carried out
at 25° C in 0.01 M phosphate buffer, 0.15 M NaCl, pH 7.

F(u^cm )
0.3

2000 4000 6000 8000

Time (s)

F i g . 6. The adsorbed amount versus time for adsorption of P~

lactoglobulin to silica followed by buffer rinsing after 1800 sec, addition of
surfactant (CTAB) after 3600 seconds and a final rinse with buffer after
5400 sec (•). Adsorption from a mixture of the protein and surfactant for
1800 sec. followed by rinsing is also included (•). Conditions as in Fig. 5.

In Proteins at Interfaces II; Horbett, T., et al.;

ACS Symposium Series; American Chemical Society: Washington, DC, 1995.
 ARNEBRANT & WAHLGREN Protein-Surfactant Interactions

r(pgtm ) 2

1.0 i
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1 1 1 1 1
o.o
0 2000 4000 6000 8000
Time (s)

Fig. 7. The adsorbed amount versus time for adsorption of fibrinogen to

silica followed by buffer rinsing after 1800 sec, addition of surfactant
(DTAB) after 3600 seconds and a final rinse with buffer after 5400 sec. (A).
Adsorption from a mixture of the protein and surfactant for 1800 sec.
followed by rinsing is also included ( • ) . Conditions as in Fig. 5.

1
0.0—• '—'—'—'—•—•—
0 2000 4000 6000 8000
Time (s)
Fig. 8. The adsorbed amount versus time for adsorption of lysozyme to
silica followed by buffer rinsing after 1800 sec, addition of surfactant
(CTAB) after 3600 seconds and a final rinse with buffer after 5400 sec. ((•).
Adsorption from a mixture of the protein and surfactant for 1800 sec.
followed by rinsing is also included (•). Conditions as in Fig. 6.

In Proteins at Interfaces II; Horbett, T., et al.;

ACS Symposium Series; American Chemical Society: Washington, DC, 1995.
 250 PROTEINS AT INTERFACES II
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P r o t e i n
Surfactant After
Adsorption Adsorption Rinse

Fig. 9. A n illustration of the different classes of surfactant -protein

interactions at solid interfaces.

components normally have higher surface activity (superiority in lowering interfacial

tension).
Experimentally it is observed that the presence of surfactants in protein solutions
may influence the amount of proteins adsorbed to solid surfaces in three different
ways (Figs. 5,7, 8):

i) Complete hindrance of protein adsorption, Fig. 5.

ii) Reduced amounts adsorbed compared to adsorption from pure protein solution,
Fig. 8.
iii) Increased amounts adsorbed, Fig. 7.

In case (i), the complete lack of adsorption could be explained:

In Proteins at Interfaces II; Horbett, T., et al.;

ACS Symposium Series; American Chemical Society: Washington, DC, 1995.
 17. ARNEBRANT & WAHLGREN Protein-Surfactant Interactions 251

1) If a complex is formed with a surfactant that has no attraction to the surface.

2) If the surfactants adsorb, due to their higher surface activity and diffusivity, and
by doing so prevent further adsorption of proteins or protein/surfactant com­
plexes.

In (ii) and (iii), the formation of complexes in solution leads to a decrease or in­
crease in the amounts of protein adsorbed, respectively. The presence of surfactant
influences the total amount of protein by steric effects or changes in electrostatic in­
teraction between complexes as opposed to native protein. In addition, the complex
may adsorb in a different orientation than the pure protein. The surfactant molecules
will desorb upon dilution, leaving protein molecules adsorbed at the surface.
Due to the different shapes of the adsorption isotherms of surfactants and
proteins, the interfacial interaction is of course, as for protein mixtures, strongly
dependent on the concentration of the components. The special character of the
surfactant adsorption isotherms featuring the sharp increase in adsorbed amount in the
range of their critical association concentration will influence these events in a very
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pronounced way. This fact and the reversibility upon dilution will give surfactant-
protein interactions much of their uniqueness. Studies regarding these surfactant-
protein "Vroman effects" have been reported, for example adsorption of fibrinogen
from mixtures with Triton X-100 has been seen to go through a maximum (63). The
adsorption from p-lactoglobulin/SDS mixtures at different degrees of dilution was
studied by Wahlgren and Arnebrant (62) (Fig. 10). At concentrations above the cmc
for the surfactant the amount adsorbed corresponded to a layer of pure surfactant and
was found to increase after rinsing. At lower concentrations, the adsorbate prior to
rinsing appeared to be a mixture of protein and surfactant and the total amount
adsorbed passes a maximum. The composition of the adsorbate after rinsing is most
likely pure p-lactoglobulin, as interactions between surface or protein and SDS are
reversible.
At high degrees of dilution of the mixture, the absence of surfactant adsorption
to the methylated silica, the non-reversible adsorption of P-lactoglobulin and the
observed partial desorption of the adsorbate from the mixture, implies that some SDS
molecules are bound to the P-lactoglobulin molecules with a higher affinity than to
the surface (Fig. 10). The amount of protein adsorbed is larger, even after rinsing,
than for adsorption from pure P-lactoglobulin solutions, and it can be concluded that
SDS- binding facilitates the adsorption of protein.
Generally, it can be concluded that surfactants may interact through solubiliza­
tion or replacement mechanisms depending on surfactant- surface interactions and
surfactant- protein binding. Solubilization requires complex formation between
protein and surfactant, and replacement adsorption of the surfactant to the surface. As
previously concluded for protein adsorption, one of the most important protein prop­
erties affecting the elutability appear to be the conformational stability. Differences
between a competitive situation and addition of surfactant after protein adsorption
may be found in the alteration in surface activity of protein- surfactant complexes
formed in solution as compared to pure protein, the difference in diffusivity of surfac­
tants and proteins effecting the "race for the interface" and time dependent confor­
mational changes resulting in "residence time" effects.
The effects at low surfactant concentration, below the cmc, involving the range
of specific binding to some proteins are not fully understood. Further, the exact pre­
requisites for solubilization versus replacement as well as detailed information on
molecular parameters such as aggregation numbers are not known. Several of the
phenomena discussed in this paper resemble those observed in polymer- surfactant
systems (64). A complete survey of the analogies is however beyond the scope of this

In Proteins at Interfaces II; Horbett, T., et al.;

ACS Symposium Series; American Chemical Society: Washington, DC, 1995.
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Publication Date: May 5, 1995 | doi: 10.1021/bk-1995-0602.ch017

cmc SDS

1000 100 10 1
Degree of dilution

Fig. 10. The amounts adsorbed to a methylated silica surface as a function

of degree of dilution for a mixture of p-lactoglobulin and SDS (1/5 w/w), in
phosphate buffered saline pH 7, 1=0.17. The figure shows the adsorbed
amount after 30 min of adsorption (O) and 30 min. after rinsing (+)• In
addition, the figure shows the adsorption of pure P-lactoglobulin, after 30
min of adsorption (•) and 30 min after rinsing (x). Finally, the adsorption
isotherm of SDS is inserted (A), data from (62).

review. As pointed out in the introduction, improved knowledge concerning surfac­

tant elution will be useful for estimation of protein attachment strength to surfaces,
for optimizing detergency processes and avoiding undesired adsorption in medical
applications.

Acknowledgement

The authors would like to thank Stefan Welin-Klintstrom and Joseph McGuire for
good collaboration and Hans Elwing, Per Claesson and Eva Blomberg for valuable
discussions. We also gratefully acknowledge financial support from Swedish
Research Council for Engineering Sciences (TFR).

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