Clustering of PK-trisaccharides on amphiphilic cyclodextrin reveals unprecedented affinity for the Shiga-like toxin Stx2

Article abstract of DOI:10.1039/c7cc06299k

Using amphiphilic cyclodextrin as a scaffold, the first class of P^K-glycoconjugates capable of high avidity binding to both Stx1 and Stx2 toxins in solid-phase assay formats is reported. The generated glycomicroarray effectively mimics the plas

Full text of DOI:10.1039/c7cc06299k

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Marilyn M. Olmstead, Alan L. Balch, Josep M. Poblet, Luis Echegoyenet al.
Reactivity differences of Sc
first methanofullerene derivatives of Sc
3
N@C2n (2n
=
68 and 80). Synthesis of the
3
N@D5h-C80
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k
Clustering of P -trisaccharide on Amphiphilic Cyclodextrin Reveals
Unprecedented Affinity for the Shiga-like Toxin, Stx2
Received 00th January 20xx,
Accepted 00th January 20xx
a
b
a
b
b
Ping Zhang, Eugenia Paszkiewicz, Qifang Wang, Joanna M. Sadowska, Pavel I. Kitov, David R.
b
a
Bundle* and Chang-Chun Ling*
DOI: 10.1039/x0xx00000x
www.rsc.org/
k
k
Using amphiphilc cyclodextrin as a scaffold, the first class of P - Consequently, the co-crystallization of Stx2 with P -
glycoconjugates capable of high avidity binding to both Stx1 and trisaccharide has not been successful. Recently studies showed
k
11
Stx2 toxins in solid-phase assay format is reported. The generated Stx2 unexpectedly binds to an unnatural trisaccharide, P NAc
glycomicroarray effectively mimics the plasma membrane surface , Fig. ) which prompted Jacobson et al to successfully
while discriminating binding of the two Stx toxins with obtain
(
3
1
6
b
the crystal structure of Stx2a with a related
unprecedented affinity to Stx2.
disaccharide, GlNAc-α(1→4)-Gal-β(1→OMe) (Fig
1).
a)
b)
OH
OH
O
H
PK
O
The enterohemorrhagic Escherichia coli (EHEC) O157:H7
1
Shiga-like toxins, Stx1 and Stx2 (Fig. 1). The two
HO
R'
N
K
6
Stx1
P
O
produces AB
5
S
O
OH
O
OH
toxins are responsible for acute kidney failure that can
HO
O
Site 3
HO
O
R
O
OH
N
O
OH
OH
progress to life threatening haemolytic uremic syndrome
2
HUS). The higher virulence of Stx2 makes this toxin clinically
Site 2
x
m-x
n
1. R'=OH, R=ceramide, Gb
2. R'=OH, R=O(CH ) CO2Me
3. R'=NHAc, R=O(CH ) N
2 6 3
3
Site 1
(
4. PK-Povidone conjugate
2
8
3
more relevant than Stx1 but for a variety of reasons it has
been more difficult to study. The cell surface glycolipid Gb (
c)
PK
1,
3
O
O
K
), possessing the commonly called P -trisaccharide
O
PK
O
Fig.
1
O
O
O
O
O
O
O
O
O
O
R
epitope, is recognized by the B homopentamers of both
5
4
toxins. The structural basis of this interaction has been the
O
N
N
N
O
R
R
RR
R
PK
O
N
N
N
N
N
N
O
O
O
O
O
Stx2
O
O
O
O
O
O
PK
O
N N N
N
O
O
O
R
R
O
O
focus of interest, since potent inhibitors of this interaction may
offer the potential for therapeutic intervention in an otherwise
O
OO
OO
N
N
R
O
O
K O
O
O
O
R
Site 3
O
O
N
N
N
P
O
O
O
O
RR
O
O
N
N
N
O
O
RR
5
Site 2
K O
O
limited landscape of treatment options.
O
P
O
O
Stx1 and Stx2 B5 pentamers have a common structural
O
O
PK
6
motif and location of binding sites. Solved crystal structure
-7
5 R=
R=
6
k
of Stx1 with a synthetic P -trisaccharide
Me
d)
2 (Fig. 1) revealed a
O
N
O
O
R
R
N
N
O
total of fifteen binding sites which are arrayed on one face of
7
the B5 pentamer.
K O
O
O
O
O
P
b
K
Stx1 toxin binds monomeric P -
O
Me
7 R=
R=
8
trisaccharide with a millimolar K , Despite the low affinity,
8
D
Fig 1. a) Crystal structures to show carbohydrate-binding sites (rendered in
subunit of Stx1, co-crystalized with ( ) (top, 1BOS) and AB of Stx2a,
4)-Gal-β(1→OMe) disaccharide (4M1U). b)
remarkably, Stx1 was reported to bind to cell surfaces with PyMol): B
5
2
5
co-crystallised with GlNAc-α(1
→
9
K
nanomolar avidity, as a result of multivalency. The fluidity of Structures of Gb
3
c)-d) Structures of the synthetic targets P -conjugates
(
1
) and synthetic analogs (
2
-
3
) and a P -povidone polymer (
4).
K
5
/
6
and
7/8.
the cell membrane probably allows translation of Gb3
molecules for optimal spatial positioning to engage each site,
generating maximum binding efficiency. On the other hand,
k
In ELISA assays, Stx2 bind to P -BSA glycoconjugate with
6b
very low avidity (EC50 ~2480 ng/mL), and our recent results
k
8b,10
Stx2 binds P -trisaccharide >10 times weaker than Stx1.
k
also revealed that it can only bind to a linear P -povidone
1
polymer (P PVP,
2
K
4, Fig. 1) with similar affinity. Its weak
K
binding to P -conjugates and low ligand occupancy of binding
sites in solved crystal structures is in sharp contrast to the
3
b
higher virulence of Stx2 toxin. In an effort to obtain improved
ligands for Stx detection and potentially also neutralization, we
k
elected to investigate scaffolds for display of P ligands that
5
could exploit the radial symmetry of the toxin B subunit.
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Cyclodextrins (CDs), possess
n
C -axial symmetry that glycosidated with O-propargylated alcohol 14. With the
matches the radial geometry of both Stx1 and Stx2 toxins. We lactoside 12 in hand, we carried out an enzymatic α-
k
are therefore interested in developing CD-based P - galactosylation using UDP-glucose 16 as a donor. The crude
19
K
glycoclusters to study their binding to toxins. Previously, CDs P -trisaccharide 17 was O-acetylated with acetic anhydride –
K
have been functionalized with simple ionic groups to either pyridine, to give the desired P -trisaccharide 11 in 66% yield
1
inhibit anthrax lethal toxin and others, or block the α-helix over two steps (Scheme 2).
3
1
4
barrel of Wza, an outer membrane protein responsible for
the export of bacterial capsular polysaccharides. Many CD-
1
5
based glycoclusters have also been prepared to study
multivalent interaction with legume lectins. In addition to their
radial geometry their pseudo face-to-face symmetry is unique
and advantageous for this class of supramolecular hosts, as
different functionalities can be installed at each face to obtain
functional biomaterials. Incorporation of oligosaccharides to
the primary face and aliphatic chains to the secondary face of
CDs generates a class of amphiphilic CD-based glycoclusters
that have the ability to self-assemble in water and most
importantly interacting with hydrophobic surfaces. We
envisioned that with numerous aliphatic chains at the same
planar surface, they could be immobilized on ELISA microtiter
plates, to print a glycoarray for assay of bioreceptors
possessing radial geometry. Such CD glycoclusters have not
been used to inhibit Stx1 and Stx2. If linkers of sufficient length
Scheme 1. Retrosynthetic scheme to targets 5 and 6.
K
were inserted between the CD scaffold and P -residues, the
carbohydrate ligands on the generated glycosurface should
have ample freedom to rotate and translate in order to adopt
favourable conformations when interacting with multivalent
5
B -pentamers of Stx toxins, effectively mimicking lateral
K
Scheme 2. Synthesis of P -trisaccharide (11) via an chemoenzymatic route.
diffusion of glycolipids in the cell membrane. Previously,
several groups have reported the synthesis of different classes
K
The conjugation between the fully protected P -
1
5,16
of glycoclusters based on amphiphilic CDs
and also
1
calixarene. Here we report the synthesis of two β-CD-based
K
P -trisaccharide glycoclusters (5-6, Fig. 1) containing either 14
7
trisaccharide 11 with the two per-6-azido derivatives of β-CD
(
9
and 10) is shown in Scheme
reacted with the per-6-azide-β-CD containing n-hexyl groups
) using 1.25 : 1 ratio of alkyne : azide. The desired conjugate
was isolated in 67% yield by column chromatography. In a
3. Compound 11 was first
K
n-hexyl or 14 n-dodecyl chains at the secondary face, and 7 P -
trisaccharide units at the primary face via the water-soluble
tetraethylene glycol linker.
(
9
1
8
K
similar fashion, the conjugation of P -trisaccharide 11 with the
analogous compound 10 was also successful, providing the
To estimate the affinity enhancement ascribable to
clusterization, we also designed two monomeric ligands 7-8,
K
desired conjugate 19 in 77%. Finally,
a
Zémplen
transesterification afforded the desired P -β-CD glycocluster
and in 91% and 88% yield, respectively, isolated by size-
which effectively represent one slice of the P -CD glycoclusters
respectively. At the O-2 / O-3 positions, we introduced
either two n-hexyl ( ) or two n-dodecyl groups ( ), and at the
K
5
5-6
6
7
8
K
exclusion chromatography on Sephadex LH-20.
C-6 position, we linked a P -trisaccharide in an identical
manner as the two CD conjugates. Furthermore, we blocked
both O-1 and O-4 positions of the α-D-glucopyranosyl unit with
a methyl group so that the polarity of the two compounds
K
closely mimics the two P -CD clusters.
Scheme
1 shows our retrosynthetic analysis of compounds
1
8
. The previously reported β-CD derivatives 9-10 were
5-
6
served as the key intermediates along with the per-O-
K
acetylated P -trisaccharide 11. This combination of functional
K
groups allowed us to perform the key conjugation of P -
trisaccharides to β-CD via the highly efficient copper-mediated
Huisgen 1,3-dipolar cycloaddition (“click” reaction). To obtain
K
the desired P -trisaccharide, we planned to perform an
enzymatic galactosylation from the key β-lactoside 12
K
The synthesis of P -trisaccharide (11, Scheme
.
K
Scheme 3. Conjugation of protected P -trisaccharide 11 to per-2,3-O-alkylated β-
2
) started CD per-6-azides
9
and 10 and final full deprotection.
from the per-O-benzoylated lactosyl bromide 13, which was
2
| Chem. Commun., 2017, 00, 1-3
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K
Syntheses of the two P -αGlc conjugates
7-
8
are illustrated broadening was observed (Fig. 2d), suggesting no formation of
in Scheme 4. The two key intermediates 29 and 30 were aggregates by the compound in methanol. Finally, for the
K
obtained by following a conventional route in excellent yields. analogous P -αGlc
1
possessing two n-hexyl chains, its H NMR
7
K
The last glycoconjugation step with the protected P - spectrum in D
2
O (Fig. 2a) appeared slightly broadened but
trisaccharide 11 afforded the desired conjugates 31 and 32 in nevertheless much better resolved than the other amphiphilic
almost quantitative yields. After an analogous O-deacetylation compounds, revealing its much reduced ability to self-
step, the two targets
7
and
8
were isolated in excellent yields.
assemble in water. The identities of all final glycoconjugates
and were ultimately confirmed by high-resolution
mass spectrometry (see ESI).
The biological activities of the two P -β-CD conjugates 5-6
5/
6
7/8
R'
N
3
Ph
O
O
Ph
O
O
O
O
O
O
a
or b
O
c
BzO
e
R'O
O
R
O
K
HO
HO
OOMe
R
OOMe
R
OOMe
R
OMe
R
R
20
K
as well as the two P -αGlc conjugates
7-8 were evaluated by
2
2
1. R=C
6
H13-n
23. R=C
25. R=C
6
H
13-n, R'=Br
27. R=C
6
H
H
H
H
13-n, R'=H
13-n, R'=Me
25-n, R'=H
25-n, R'=Me
d
d
f
f
2. R=C
6
H13-n
6
H
13-n, R'=N
3
29. R=C
6
ELISA using a direct binding assay format. Except for
2
2
4. R=C12
6. R=C12
H
H
25-n, R'=Br
25-n, R'=N
28. R=C12
30. R=C12
k
3
compound
7
, all other P -conjugates were found to coat the
shows the
and to
interact with both Stx1 and Stx2 and the results were
OR'
OR'
O
hydrophobic surface of ELISA plates well. Fig 3
R'O
O
a. n-C
6
H13Br, NaH/DMF;
1
1
O
O
3
K
O
R'O
O
b. n-C12H25Br, NaH/DMF;
c. NBS/BaCO
/heat;
d. NaN
results of printed surface of both P -β-CD ligands
5
6
OR'
OR'
3 2 2
/ClCH CH Cl
g
O
R'O
N
O
N
N
3
/DMF/70° C;
k
OR'
OR'
e. NaOMe/MeOH;
f. MeI/NaH/DMF;
compared with a P PVP polymer (
K
monomeric P -αGlc liand 8. The same amount (10 μg/mL, 100
4
) as well as with the
R'O
O
2
9 or 30
MeO
O
g. CuI/iPr
reflux;
2
NEt/acetone,
3
3
1. R=C
. R=C
6
6
H
H
13-n, R'=Ac
13-n, R'=H
25-n, R'=Ac
H25-n, R'=H
h
h
O
R
7
R
OMe
μL/well) of each sample was applied to the microtiter plate. As
can be seen, strikingly differences are observed between Stx1
h. NaOMe/MeOH
2. R=C12
. R=C12
H
8
K
K
and Stx2 when interacting with the two P -β-CD conjugates.
Scheme 4. Synthesis route to the amphiphilic P -αGlc monomers (7-8).
K
The two P -β-CD conjugates
5-6 clearly showed stronger
K
In the NMR spectra of all P -β-CD conjugates, an axial binding to Stx2 (~8-10 ng/mL) than Stx1 (~45-55 ng/mL). This is
symmetry is observed. For example, only one set of triazole in contrast to the monomeric ligand
8
which showed a reverse
1
signal was observed in the H NMR spectra of each conjugate; trend by binding to Stx1 (~150 ng/mL) much stronger than to
1
13
K
8b,10
additionally, for each of the H/ C signal of P -trisaccharides, Stx2 (~1900 ng/mL), which is consistent with literature.
K
only one set of NMR signal was also observed, confirming the Stx1 toxin binds to both P -β-CD conjugates
5-6
weaker than
) (10 ng/mL) but only slightly better than the
residues to the β-CD core. For the two final P -β-CD conjugates monomeric ligand
(EC50 150 ng/mL). On the other hand, Stx2
, as expected, their H signals are generally very broad in clearly prefers interacting with spatially prearranged
deuterated water (D O, Fig. 2b). This is expected for amphiphilic CD-based glycoconjugates; detection of Stx2 by
amphiphilic molecules in water because they likely form both P -β-CD conjugates
K
k
complete and uniform incorporation of P -trisaccharide P PVP polymer (
4
K
8
1
5/6
2
K
5-
6
occurs at more than two orders of
micelles of different sizes. The H spectrum of the compound magnitudes lower concentration (EC50 ~8-10 ng/mL) than the
possessing n-dodecyl chains ( ) showed even more broadness monomeric ligand
(EC50 ~1900 ng/mL), which interestingly
see ESI), revealing its stronger ability to self-assemble.
binds to Stx2 at about the same strength as the polymeric
1
6
8
(
k
P PVP (4, EC50 ~1800 ng/mL). However, it should be noted that
the above comparisons were based on presumed similar
K
coating densities of P -ligands. Considering the fact that
monomeric ligand (
plates with the same efficiency as the two P -β-CD conjugates,
the above observed increase (compared to monomer ) could
be overestimated. When comparing the P -glycoarray
constructed with the linear polymeric ligand ( ) to the one
8) is most unlikely to print the microtiter
K
8
K
4
constructed with the monomeric ligand, we note there does
not appear to exist much difference for Stx2 detection.
Furthermore, the different chain lengths attached to the two
CD-conjugates did not affect the binding significantly,
confirming that they essentially serve as anchor groups of the
ligands to the hydrophobic surface.
K
In conclusion, amiphiphilic CD-based P -glycoclusters
1
. Partial 1D H NMR spectra of a) monomeric P -αGlc
K
Fig 2
MHz), b) P -βCD glycocluster
7
(D
2
O, 298 K, 600
O, 298 K, 600 MHz), c) monomeric P -αGlc 8 in
K
K
5 and
k
reported here are the first P -glycoconjugates capable of
5
O (298 K, 400 MHz) and d) monomeric P -αGlc
(D
2
K
D
2
8
in CD
3
OD (298 K, 600 MHz).
6
high avidity binding to both Stx toxins in a solid phase assay
format. In contrast to previous solid phase assays a marked
selectivity for binding to Stx2 over Stx1 is observed by
K
For the monomeric P -αGlc conjugate
8
that contains two
O also appeared
very broad (Fig. 2c), which resembles compounds . On the
1
n-dodecyl chains, its H NMR spectrum in D
2
5/6
K
amphiphilic CD-based P -conjugates. Compared to monomeric
K
, the two P -β-CD conjugates
1
other hand, when the H NMR spectrum of the same
compound was recorded in deuterated methanol, no signal
ligands
7
and
8
5
-6
showed a
remarkable increase in Stx2 binding avidity, presumably due to
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K
Miyazawa, C. Yamasaki, J. Aoki, S. Takashima, Y. Yamakawa,
M. Nishijima, D. Terunuma, H. Kuzuhara and Y. Natori, Proc.
Natl. Acad. Sci. U. S. A., 2002, 99, 7669; d) A. A. Kulkarni, C.
Fuller, H. Korman, A. A. Weiss and S. S. Iyer Bioconjugate
Chem., 2010, 21, 1486.
a) M. E. Fraser, M. Fujinaga, M. M. Cherney, A. R. Melton-
Celsa, E. M. Twiddy, A. D. O' Brien and M. N. G. James, J. Biol.
Chem., 2004, 279, 27511; b) J. M. Jacobson, J. Yin, P. I. Kitov,
G. Mulvey, T. Griener, M. James, G. D. Armstrong and D. R.
Bundle, J. Biol. Chem., 2014, 289, 885.
the preclustering of P -ligands on the radial scaffold that
provides secondary preorganization. The exact magnitude of
this avidity increase requires
a rigorous quantitative
comparison using a family of newly designed ligands for
solution phase assays. Nevertheless, the results from this work
shows that prearrangement of carbohydrate ligands on CDs as
natural circular scaffolds constitutes an interesting new way of
presenting specific ligands to certain biological receptors that
have complementary geometry. In solid-phase assay, such
platforms may mimic the presentation of glycolipids in cell
membrane. Biosensor applications and array screening
platforms can be envisioned.
6
7
8
a) P. E. Stein, A. Boodhoo, G. J. Tyrrell, J. L. Brunton and R. J.
Read, Nature, 1992, 355, 748; b) H. Ling A. Boodhoo, B.
Hazes, M. D. Cummings, G. D. Armstrong, J. L. Brunton and R.
J. Read, Biochemistry, 1998, 37, 1777.
a) P. M. St. Hilaire, M. K. Boyd and E. J. Toone, Biochemistry,
1
994, 33, 14452-14463; b) E. N. Kitova, P. I. Kitov, E.
Paszkiewicz, J. Kim, G. L. Mulvey, G. D. Armstrong, D. R.
Bundle and J. S. Klassen, Glycobiology, 2007, 17, 1127.
S. C. Head, M. A. Karmali and C. A. Lingwood, J. Biol. Chem.,
9
1
1
1
991, 266, 3617.
0 M. Mylvaganam and C. A. Lingwood, Biochem. Biophys. Res.
Commun., 1999, 257, 391.
1 R. R. Kale, C. M. McGannon, C. Fuller-Schaefer, D. M. Hatch,
M. J. Flagler, S. D. Gamage, A. A. Weiss and S. S. Iyer, Angew.
Chem., Int. Ed., 2008, 47, 1265.
1
1
2 P. I. Kitov, Y. Kotsuchibashi, E. Paszkiewicz, D. Wilhelm, R.
Narain and D. R. Bundle, Org. Lett., 2013, 15, 5190.
3 a) V. A. Karginov, E. M. Nestorovich, M. Moayeri, S. H. Leppla
and S. M. Bezrukov, Proc. Natl. Acad. Sci. U. S. A., 2005, 102
5075; b) V. A. Karginov, A. Yohannes, T. M. Robinson, N. E.
Fahmi, K. Alibek and S. M. Hecht, Bioorg. Med. Chem., 2006,
, 33; c) M. Moayeri, T. M. Robinson, S. H. Leppla and V. A.
,
1
1
4
Karginov, Antimicrob. Agents Chemother., 2008, 2239; d) E.
M. Nestorovicha, V. A. Karginov, A. M. Berezhkovskii, V. A.
Parsegian and S. M. Bezrukov, Proc Natl Acad Sci U S A, 2012,
1
09, 18453 and the references therein; e) M. Roeder, E. M.
Nestorovich, V. A. Karginov, C. Schwan, K. Aktories and H.
Barth, Toxins, 2014, , 2097; f) A. Díaz-Moscoso, A. Méndez-
6
Ardoy, F. Ortega-Caballero, J. M. Benito, C. Ortiz Mellet, J.
Defaye, T. M. Robinson, A. Yohannes, V. A. Karginov, J. M.
García Fernández, ChemMedChem, 2011,
4 L. Kong, L. Harrington, Q. Li, S. Cheley, B. G. Davis and H.
Bayley, Nat. Chem., 2013, , 651.
15 For reviews: see: a) A. Martinez, C. Ortiz Mellet and J. M.
Garcia Fernandez, Chem. Soc. Rev., 2013, 42, 4746 and
references therein. b) F. Sallas and R. Darcy, Eur. J. Org.
Chem., 2008, 957 and references therein.
6, 181.
1
5
K
Titration of Stx1 (top) and Stx2 (bottom) on plates coated with P -βCD
Fig 3.
glycoclusters
K
6, and compared with the P -αGlc monomeric conjugate 8 as
5 and
well as P -povidone (P PVP) polymer (
K
k
4).
We acknowlege the financial support of Alberta Innovates
Technology Futures and the University of Calgary.
1
6 a) A. Mazzaglia, D. Forde, D. Garozzo, P. Malvagna, B. J.
Ravoo and R. Darcy, Org. Biomol. Chem., 2004, , 957; b) S.
McNicholas, A. Rencurosi, L. Lay, A. Mazzaglia, L. Sturiale, M.
Perez, and R. Darcy, Biomacromolecules, 2007, , 1851. c) A.
–
2
8
Notes and references
Díaz-Moscoso, N. Guilloteau, C. Bienvenu, A. Méndez-Ardoy,
J. L. Jiménez Blanco, J. M. Benito, L. Le Gourriérec, C. Di
Giorgio, P. Vierling, J. Defaye, C. Ortiz Mellet and J. M. García
Fernández, Biomaterials, 2011, 32, 7263; d)F. Sallas, K.
Nikura and S.-I. Nishimura, Chem. Commun., 2004, 596.
7 R. Roy and J. M. Kim, Angew. Chem., Int. Ed., 1999, 38, 369
8 H. Parrot-Lopez, C.-C. Ling, P. Zhang, A. Baszkin, G. Albrecht,
C. de Rango and A. W. Coleman, J. Am. Chem. Soc., 1992,
1
2
M. A . Karmali, Clin. Microbiol. Rev., 1989,
P. I. Tarr, C. A. Gordon and W. L. Chandler, Lancet, 2005, 365
073.
2, 15.
,
1
3
4
C. A Fuller, C. A Pellino, M. J. Flagler, J. E. Strasser and A. A.
Weiss, Infect. Immun., 2011, 79, 1329.
a) A. Cohen, G. E. Hannigan, B. R. Williams and C. A.
Lingwood, J Biol Chem., 1987, 262, 17088; b) T. Waddell, S.
Head, M. A. Petric, A. Cohen and C. A. Lingwood, Biochem.
Biophys. Res. Commun., 1988, 152, 674.
1
1
1
14, 5479-5480.
9 P. I. Kitov. E. Paszkiewicz, W. W Wakarchuk and D. R. Bundle,
1
Methods Enzymol., 2003, 362, 865.
5
a) G. D. Armstrong, P. C. Rowe, P. Goodyear. Elaine Orrbine,
T. P. Klassen, G. Wells, A. MacKenzie, H. Lior, C. Blanchard, F.
Auclair, B. Thompson, D. J. Rafter and P. N. McLaine, J. Infect.
Dis., 1995, 171, 1042; b) P.I. Kitov, J. M. Sadowska, G.
Mulvey, G. D. Armstrong, H. Ling, N. S. Pannu, and R. J. Read,
D. R. Bundle, Nature, 2000, 403, 669; c) K. Nishikawa, K.
Matsuoka, E. Kita, N. Okabe, M. Mizuguchi, K. Hino, S.
4
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DOI: 10.1039/C7CC06299K
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Amphiphilic cyclodextrin-based P -glycoarrays show remarkable binding avidity and selectivity for Stx2 in
solid phase assay formats