Multivalent neuraminidase hydrolysis resistant triazole-sialoside protein conjugates as influenza-adsorbents

Article abstract of DOI:10.1016/j.cclet.2017.10.032

We report the synthesis of pseudo triazole-sialoside protein conjugates of various valency that are resistant to neuraminidase for the adsorption of influenza viruses. The glycotriazole monomer bearing an amine-functionalized linker was synthesized by click chemistry and grafted to the lysine residues of bovine serum albumin (BSA) or human serum albumin (HSA) via diethyl squarate and adipate-based strategy. The binding of hemagglutinin (HA) and neuraminidase (NA) on the virion surface by the synthetic neoglycoproteins were evaluated by hemagglutination and neuraminidase inhibition assay, respectively. The results demonstrated that these synthetic glycoproteins have significantly higher affinity with NA than HA. The interactions between these neoglycoproteins and intact influenza viruses were further investigated by Dynamic Light Scattering (DLS) technique. The pronounced agglutination indicated that these glycoconjugates can be used as adsorbents to prevent virus from invading host cells as well as the release of newly synthesized viral particles, which are crucial in the life cycle of the influenza virus. With the high binding affinity to intact influenza viruses, these neoglycoproteins can also be used as probe to elucidate the molecular mechanism of the sialic acid-influenza recognition and biosensors for influenza detection.

Full text of DOI:10.1016/j.cclet.2017.10.032

G Model
CCLET 4305 No. of Pages 5
Chinese Chemical Letters xxx (2017) xxx–xxx
Contents lists available at ScienceDirect
Chinese Chemical Letters
journal homepage: www.elsevier.com/locate/cclet
Communication
Multivalent neuraminidase hydrolysis resistant triazole-sialoside
protein conjugates as influenza-adsorbents
a
a
a
a
a
a
Xin Menga, , Meibing Yang , Yang Li , Xiaobin Li , Tianwei Jia , Haojie He , Qun Yu ,
*
Na Guoa, Yun Heb, Peng Yua, , Yang Yang
*
China International Science and Technology Cooperation Base of Food Nutrition/Safety and Medicinal Chemistry, College of Biotechnology, Tianjin University
of Science and Technology, Tianjin 300457, China
a,
*
a
b
Research Institute of Tsinghua University in Shenzhen, Shenzhen 518057, China
A R T I C L E I N F O
A B S T R A C T
Article history:
We report the synthesis of pseudo triazole-sialoside protein conjugates of various valency that are
resistant to neuraminidase for the adsorption of influenza viruses. The glycotriazole monomer bearing an
amine-functionalized linker was synthesized by click chemistry and grafted to the lysine residues of
bovine serum albumin (BSA) or human serum albumin (HSA) via diethyl squarate and adipate-based
strategy. The binding of hemagglutinin (HA) and neuraminidase (NA) on the virion surface by the
synthetic neoglycoproteins were evaluated by hemagglutination and neuraminidase inhibition assay,
respectively. The results demonstrated that these synthetic glycoproteins have significantly higher
affinity with NA than HA. The interactions between these neoglycoproteins and intact influenza viruses
were further investigated by Dynamic Light Scattering (DLS) technique. The pronounced agglutination
indicated that these glycoconjugates can be used as adsorbents to prevent virus from invading host cells
as well as the release of newly synthesized viral particles, which are crucial in the life cycle of the
influenza virus. With the high binding affinity to intact influenza viruses, these neoglycoproteins can also
be used as probe to elucidate the molecular mechanism of the sialic acid-influenza recognition and
biosensors for influenza detection.
Received 29 June 2017
Received in revised form 7 August 2017
Accepted 23 October 2017
Available online xxx
Keywords:
Click chemistry
Multivalent effect
Glycoprotein
Dynamic light scattering
Hemagglutinin/Neuraminidase inhibitor
© 2017 Chinese Chemical Society and Institute of Materia Medica, Chinese Academy of Medical Sciences.
Published by Elsevier B.V. All rights reserved.
N-Acetyl neuraminic acid (Neu5Ac, also called sialic acid, SA)
(Scheme 1), an acidic monosaccharide with nine-carbon backbone,
is commonly found on the termini branches of N-glycans, O-
glycans, glycosphingolipids and glycoproteins on the cell surface
[1]. Due to its external position, SA is fully accesible to other
biomolecules and further control cell-cell interactions [2]. It has
been found that SA-protein interactions play vital roles in various
biological and pathological processes including cancer [3,4],
inflammation [5] and immunization [6]. Moreover, many viruses
[7,8] and bacteria [9,10] also utilized this interaction at various
stages in their life cycles for cell entry or release.
multivalent SA-protein interactions serve important roles in the
initial and final steps of the replication cycles of influenza viruses
[12]. Hemagglutinin (HA) is one of the major surface glycoprotein
of the virus (80%, ꢀ300 copies of trimer), which binds to
a-SA to
induce fusion between viral and cellular membranes [13].
Neuraminidase (NA) is another receptor-destroying surface
glycoprotein (17%, ꢀ50 copies of tetramer), it cleaves the residual
SA to release the virus from infected cells [14]. Two FDA approved
SA derivatives Zanamivir and Osetamivir as potent NA inhibitors
were invented based on the elucidation of SA-NA interactions,
which were effective measurements to prevent potential Flu
pandemics [15]. However, the mutations lead to the drug
resistance [16] have decreased the effectiveness of the two drugs,
thus developing new anti-influenza agents are in great need.
An alternative strategy for the development of antiviral agents
is inspired by the mucin [17], a heavily glycosylated protein
secreted by epithelial tissues of organisms of mammal to trap
viruses and expel the virions by mucocilliary transportation via
multivalent SA-HA/NA interactions [18]. Native O-linked sialoside
can be hydrolyzed by NA [19], which is the major drawback of using
Influenza virus, a genus of the Orthomyxoviridae family that
causes outbreaks of respiratory disease as annual epidemics and
unpredictable pandemics remains a significant risk to global health
and economy [11]. It has been widely accepted that two
* Corresponding authors.
E-mail addresses: mengx@tust.edu.cn (X. Meng), yupeng@tust.edu.cn (P. Yu),
yyang@tust.edu.cn (Y. Yang).
https://doi.org/10.1016/j.cclet.2017.10.032
1001-8417/© 2017 Chinese Chemical Society and Institute of Materia Medica, Chinese Academy of Medical Sciences. Published by Elsevier B.V. All rights reserved.
Please cite this article in press as: X. Meng, et al., Multivalent neuraminidase hydrolysis resistant triazole-sialoside protein conjugates as
influenza-adsorbents, Chin. Chem. Lett. (2017), https://doi.org/10.1016/j.cclet.2017.10.032

G Model
CCLET 4305 No. of Pages 5
2
X. Meng et al. / Chinese Chemical Letters xxx (2017) xxx–xxx
Scheme 1. Synthesis of di- and tetra-valent triazole-sialoside. (a) (i) H+ resine/MeOH, (ii) Ac2O/Py. (b) BiCl3/CH3SiCl3, CH2Cl2, 90%. (c) NaN3/nBu4HSO4, CH2Cl2/H2O. (d)
CuSO4 5H2O, VcNa, THF/H2O, alkynyl scaffold. (e) (i) CH3ONa/CH3OH, (ii) NaOH, H2O/MeOH.
Á
it directly as virus inhibitor. Alternatively, presenting multivalent
NA resistant pseudo-SA on different scaffolds including polymer
[20], liposome [21], dendrimers [22] and nanoparticles [23] as
mucin mimic to bound to both HA and NA have been developed as
the virus adsorbents to prevent the infection. Our previous work
[24] has also demonstrated that unlike natural O-sialylated
complex-type glycan protein conjugates [25], NA resistant S-
sialosides protein conjugates can bind not only to HA with high
affinity resulting in the inhibition of the viral adhesion to
erythrocytes, but also NA with moderate affinity resulting in the
prevention of the hydrolysis of the SA to reduce virus propagation.
Compared with other SA modified macromolecules, the sialyl
Scheme 2. Synthesis of multivalent triazole-sialoside and PEG protein conjugates. (a) CuSO4 5H2O, VcNa, THF/H2O. (b) (i) CH3ONa/CH3OH, (ii) NaOH, H2O/MeOH. (c) TFA/
Á
CH2Cl2. (d) Squaric acid diethyl esters, phosphate buffer saline (pH 7.0). (e) BSA/HSA, borate buffer buffer (pH 9.0). (f) (i) Et3N, DMSO, (ii) sodium phosphate buffer (pH 7.5),
overnight. (g) p-Toluensulfonyl chloride, TosCl/Et3N. (h) (i) NaN3/DMF, (ii) H2, Pd(OH)2/C.
Please cite this article in press as: X. Meng, et al., Multivalent neuraminidase hydrolysis resistant triazole-sialoside protein conjugates as
influenza-adsorbents, Chin. Chem. Lett. (2017), https://doi.org/10.1016/j.cclet.2017.10.032

G Model
CCLET 4305 No. of Pages 5
X. Meng et al. / Chinese Chemical Letters xxx (2017) xxx–xxx
3
human serum albumin (HSA) and bovine serum albumin (BSA) can
closely mimic the 3D natural presentation of SA on the mucin
protein scaffold. Moreover, the density of the SA on the protein
surface can be easily determined by matrix-assisted laser
desorption ionization time of flight mass spectrometry (MALDI-
TOF-MS), which makes it feasible to quantitatively analyze the SA-
HA/NA interactions. Finally, these glycoconjugates also showed
negligible cytotoxicity against human cells with concentration up
shown in Scheme 2, mass analysis of glycoconjugates revealed that
an average of 7 SA residues with OEG linker were loaded onto one
molecule of BSA or HSA (see supporting information). Methox-
ypolyethylene glycols (HO-PEG-OMe, MW ꢀ 2000) modified BSA,
BSA-PEG-OMe and HSA, HSA-PEG-OMe were also prepared with
adipate linker as controls.
With the glycoconjugates in hand, we investigated the binding
between the synthetic neoglycoproteins and influenza virus. The
adsorption activity was first evaluated by Hemagglutination
Inhibition (HAI) assay [31]. HA can bind to the SA on the surface
of chicken red blood cells (CRBCs) resulting in agglutination, which
can be inhibited by the adsorption of HA by our synthetic
glycoconjugates. The lowest concentration of the conjugates to
prevent an influenza virus induced hemagglutination was mea-
sured and defined as Ki [32]. Three strains of influenza virus [A/
Puerto Rico/8/1934 (H1N1), A/Huairou Beijing/11069/2014 (H3N2),
A/Chicken/Beijing/AT609/2014 (H9N2)] with different HA subtype
were chosen. Mucins from bovine submaxillary glands (BSG) were
used as positive control due to its high sialyled protein content
[33].
to 100 mmol/L. These initial results encourage us to develop non-
hydrolyzable pseudo-sialoside protein conjugates library with
different HA/NA binding affinity pattern to the influenza virus
and further investigate the recognition and interaction process of
sialoside to HA or NA.
Herein, a new class of triazole-sialoside protein conjugates was
prepared and their influenza adsorption activity was evaluated by
hemagglutinin/neuraminidase inhibition. Furthermore, the bind-
ing affinity of the glycoconjugates to intact virus was studied by the
dynamic light scattering (DLS). We wish with other sensor
technology, these triazole-sialoside protein conjugates could
directly serve as anti-influenza agent development and capture
molecules for influenza to complement the elucidation of the SA-
HA/NA molecular recognition and interaction for the detection of
influenza viruses.
We found that all of the monomeric SA, low valent triazole-
sialosides (DT and TT) and BSA had no inhibitory effect on the
hemagglutination with the concentration up to 1 mmol/L (Table 2).
Regarding the glycoconjugates, only the BSA-SA2 and HSA-SA4
inhibited hemagglutination at concentrations around 50-
At first, the key intermediate peracetylated 2-azido-
acid 3 was synthesized using the method shown in Scheme 1. Fully
protected SA was first converted to -chloride derivative 2 with a
a-sialic
b
100 mmol/L when H3N2 and H9N2 were used as the sources of
catalytic amount of BiCl3 in the presence of CH3SiCl3 in CH2Cl2 [26].
A phase-transfer catalysis process was used to introduce azido
group on C-2 position with a-configuration, which was confirmed
by the NMR spectra [27] (see Supporting information). Then click
chemistry [28,29] was employed to attach the SA monomer to the
alkynyl scaffolds to afford fully protected di- and tetravalent
HA. Although BSA-OEG-SA7 and HSA-OEG-SA7 have higher density
of triazole-sialoside residue on the protein surface, no enhanced
hemagglutination inhibition was observed. This clearly demon-
strated that appropriate dimensional presentation of the sugar
ligand on the surface of the scaffold is the key factor for the binding
affinity enhancement of multivalent carbohydrate-protein inter-
action [34]. We speculated that the distribution of the SA on the
protein surface in BSA-OEG-SA7 and HSA-OEG-SA7 was not
homogeneous, which couldnot fit the exposed binding site of
HA trimers and resulted in decreased binding to the HA. On the
other hand, compared with the S-sialoside protein conjugates
prepared by us previously, the triazole-sialosides protein con-
jugates BSA-OEG-SA7 and HSA-OEG-SA7 were found to show
worse inhibition performance due to the different triazole
structure on anomeric centre. It is therefore envisioned that
different pseudo-SA conjugates have a potential application to
differentiate and classify various influenza virus strains due to
their different binding affinity to HA, which have been already
initially proved by our group [35]. When PR8 strain was used as the
HA source, no hemagglutination inhibition was observed due to
the lack of galactosyl linkage to the SA in all of the glycoconjugate
which is in agreement with our [24] and other group’s results [33].
Because all types of SA linkages are contained in the BSG mucins, it
triazole-sialoside
4 and 5. After deacylation, demethylation,
purification on Sephadex LH-20 and lyophilisation under standard
procedures, di-, DT and tetravalent, TT triazole-sialoside were
prepared, respectively (Scheme 1).
The neoglycoprotein conjugates BSA-SA2 and HSA-SA4 were
synthesized using the dimethyl squarate strategy as we previously
reported [24] (Scheme 2). The molecular weights of the resulting
glycoconjugates were characterized by MALDI-TOF-MS (see
supporting information). Unfortunately, high density of pseudo
SA modification was not achieved as the S-sialoside BSA or HSA
conjugates we prepared previously. Mass analysis of the glyco-
conjugates (BSA-SA2 and HSA-SA4) revealed that only about two or
four triazole-sialosides were attached on one molecule of BSA or
HSA, respectively, even when increasing the starting reaction
molar ratio of 8 and protein to 50:1 (Table 1). This could due to the
steric hindrance effect caused by the triazole ring on 2-position of
SA, which reduce the protein coupling efficiency. In order to attach
more sialoside residue on the protein scaffold, longer and more
flexible spacer arm (polyethylene glycol, n = 3, OEG) and N-
hydroxysuccinimide (NHS)-activated adipate [30] were adopted
to covalently couple primary amine of the triazole-SA to lysine
residues on protein. After the synthesis and purification steps as
Table 2
Inhibition constant Ki of HAI assay.
Entry
Ki (mmol/L)
H1N1
H3N2
H9N2
SA
DT
TT
>1000
NAa
NA
>1000
NA
NA
>1000
NA
NA
Table 1
The conjugation of triazole-sialoside with proteins.
BSA
>1000
NA
NA
125
NA
>1000
50
62.5
125
100
NA
>1000
50
62.5
125
100
NA
Entry
Molecular weighta
Loading SA residues
BSA-SA2
BSA-OEG-SA7
HSA
HSA-SA4
HSA-OEG-SA7
Mucin
BSA
66612.42
67698.92
71320.31
66714.25
68759.90
71519.56
0
2
7
0
4
7
BSA-SA2
BSA-OEG-SA7
HSA
HSA-SA4
HSA-OEG-SA7
NA
0.16 (1.95b)
0.16
0.16
a
No Activity.
Data reported from Ref [33].
a
b
Molecular weight was determined by MALDI-TOF-MS.
Please cite this article in press as: X. Meng, et al., Multivalent neuraminidase hydrolysis resistant triazole-sialoside protein conjugates as
influenza-adsorbents, Chin. Chem. Lett. (2017), https://doi.org/10.1016/j.cclet.2017.10.032

G Model
CCLET 4305 No. of Pages 5
4
X. Meng et al. / Chinese Chemical Letters xxx (2017) xxx–xxx
Table 3
IC50 values in the neuramidinase inhibition assay.
Entry
IC50 (mmol/L)
H1N1
H3N2
H9N2
SA
DT
TT
BSA
BSA-SA2
BSA-OEG-SA7
HSA
HSA-SA4
HSA-OEG-SA7
Zanamivir
>250
25
>250
11
NA
>500
NA
NA
>250
222
88
>250
7
NA
>500
NA
NA
>250
203
56
NAa
>500
NA
NA
>250
187
38
4 nmol/L
2 nmol/L
2 nmol/L
a
No Activity.
showed potent inhibition toward all virus strains as positive
control.
Fig. 2. Cytotoxicity study of glycoproteins.
We next performed the fluorescent MUNANA [2'-(4-methyl-
umbelliferyl)-a-D-N-acetylneuraminic acid]-based NA inhibition
in PBS before and after mixing with H9N2 was measured and the
intensity-based size distribution was plotted in Fig. 1a. The
diameters of the different molecule grafted protein and H9N2
virus in solution were found to be around 15 and 150 nm,
respectively. After the addition of the virus to the sialyl modified
HSA, DLS indicated that the mean diameter of the conjugate had
increased to 250 nm, indicating the agglutination [42] formed by
the multivalent interaction between the HSA-OEG-SA7 and H9N2.
On the contrary, the addition of the virus did not increase the size
of the HSA-PEG-OMe, as still two separated peaks at 15 nm and
150 nm, which were assigned to individual HSA-PEG-OMe and
viron, respectively. Similar results were observed when H3N2 and
H1N1 were added to the conjugates solution (Fig. 1b and c). All of
the DLS size distribution curves clearly showed our glycoconjugate
can specifically attach to the surface of the influenza virus and
could also cross-link several virus particles through multivalent
SA-NA interactions. This glycoconjugate is promising candidate for
more effective viral shielding agent in antiviral therapies and
influenza detection development with other detection technolo-
gies.
assay [36] to evaluate the NA binding activity of our glycocon-
jugtes. The fluorescent signal produced by the hydrolysis of
MUNANA by NA decreased when the glycoprotein conjugates
competitively bind to the active site of NA compared to blank. In
this assay system, the same virus strains inactivated by
b-propio-
lactone ( -PL) as mentioned in the HAI assay were used as the
b
sources of NAs. The IC50 was calculated as the concentration of the
glycoconjugates resulting in a 50% reduction in hydrolysis reaction
rate compared to the blank and Zanamivir was used as positive
control [37] (Table 3).
The data showed that divalent SA derivative is significantly
more potent than monomeric and tetravalent sialoside against all
the three strains with IC50 around 10 mmol/L which is in the range
of literature values [27]. Interestingly, TT, HSA-SA4 and even HSA-
OEG-SA7 have more sialoside residue per molecule did not
enhance the inhibitory activity. These could be explained by the
incorrect presentation of SA on the surface of the backbone which
cannot achieve effective interactions with the NA active site on the
same virus particle or adjacent other NA in different virions [38].
When changing the protein backbone from BSA to HSA, both of the
glycoproteins HSA-SA4 and HSA-OEG-SA7 have the best inhibition
activity against all virus strains, which is consistent with our
previous results [24]. More importantly, these results also
indicated SA protein conjugate for multivalent carbohydrate
display [39] and appropriate dimensional presentations on protein
backbone are important in the development of NA inhibitor and
native mucins mimic.
Finally, it was essential to study the cell toxicity of the
neoglycoproteins for the antiviral applications. The NCI-H1299,
human non-small cell lung cancer cells were incubated in the
presence of glycoproteins for 24 h. The cell activity was determined
by MTT [43]. Fig. 2 showed that the cell viability was maintained at
nearly to 80% compared to the untreated blank control under the
concentration of 250 mmol/L, indicating the extensive medical
developments of these glycoconjugates.
The influenza adsorbent behaviours of the neoglycoproteins,
HSA-OEG-SA7 which have the highest sugar density and NA
binding activity were further investigated by dynamic light
scattering (DLS) in solution [40,41]. The size of the glycoconjugates
To a summary, we have successfully synthesized multivalent
hydrolysis resistant triazole-sialoside monomer by click chemistry
and their protein conjugates via squaric acid diester and adipate
NHS ester strategy. The prepared glycoconjugates can bound to HA
Fig. 1. Hydrodynamic size distribution curves of the influenza virus a) Influenza A/Chicken/Beijing/AT609/2014 (H9N2). b) Influenza A/Puerto Rico/8/1934 (H1N1). c)
Influenza A/Huairou, Beijing/11069/2014 (H3N2) before and after mixed with different molecule modified HSA.
Please cite this article in press as: X. Meng, et al., Multivalent neuraminidase hydrolysis resistant triazole-sialoside protein conjugates as
influenza-adsorbents, Chin. Chem. Lett. (2017), https://doi.org/10.1016/j.cclet.2017.10.032

G Model
CCLET 4305 No. of Pages 5
X. Meng et al. / Chinese Chemical Letters xxx (2017) xxx–xxx
5
to prevent the viral adhesion to erythrocytes. Moreover these
glycoproteins can also strongly bind to NA to inhibit the hydrolysis
of the sialylside for the virus releasing. In addition, a pronounced
aggregation can also be observed by DLS when mixing the sialoside
protein conjugates with influenza virus through multivalent
interactions. Our findings suggest these pseudo-sialoside protein
conjugates could serve as effective influenza virus capture in
biosensor application, detection and antiviral development.
[10] S.J. Crennell, E.F. Garman, W.G. Laver, E.R. Vimr, G.L. Taylor, Proc. Natl. Acad. Sci.
U. S. A 90 (1993) 9852–9856.
[11] R.G. Webster, W.J. Bean, O.T. Gorman, T.M. Chambers, Y. Kawaoka, Microbiol.
Rev. 56 (1992) 152–179.
[12] R. Wagner, M. Matrosovich, H.D. Klenk, Rev. Med. Virol. 12 (2002) 159–166.
[13] J.J. Skehel, D.C. Wiley, Annu. Rev. Biochem. 69 (2000) 531–569.
[14] P.M. Colman, Protein Sci. 3 (1994) 1687–1696.
[15] L.V. Gubareva, L. Kaiser, F.G. Hayden, Lancet 355 (2000) 827–835.
[16] A. Moscona, N. Engl. J. Med. 353 (2005) 2633–2636.
[17] S.K. Linden, P. Sutton, N.G. Karlsson, V. Korolik, M.A. McGuckin, Mucosal
Immunol. 1 (2008) 183–197.
[18] O. Lieleg, C. Lieleg, J. Bloom, C.B. Buck, K. Ribbeck, Biomacromolecules 13
Acknowledgments
(2012) 1724–1732.
[19] J.C. Wilson, M.J. Kiefel, D.I. Angus, M. von Itzstein, Org. Lett. 1 (1999) 443–446.
[20] A. Nazemi, S.M.M. Haeryfar, E.R. Gillies, Langmuir 29 (2013) 6420–6428.
[21] H.W. Yeh, T.S. Lin, H.W. Wang, et al., Org. Biomol. Chem.13 (2015) 11518–11528.
[22] D. Zanini, R. Roy, J. Org. Chem. 63 (1998) 3486–3491.
[23] F. Feng, Y. Sakoda, T. Ohyanagi, et al., Antiviral Chem. Chemother. 23 (2013) 59–
65.
[24] Y. Yang, H.P. Liu, Q. Yu, et al., Carbohydr. Res. 435 (2016) 68–75.
[25] H. Wang, W. Huang, J. Orwenyo, et al., Bioorg. Med. Chem. 21 (2013) 2037–
2044.
[26] J.L. Montero, J.Y. Winum, A. Leydet, et al., Carbohydr. Res. 297 (1997) 175–180.
[27] M. Weïwer, C.C. Chen, M.M. Kemp, R.J. Linhardt, Eur. J. Org. Chem. 2009 (2009)
2611–2620.
[28] W.Q. Zhang, Y. He, Q. Yu, et al., Eur. J. Med. Chem. 121 (2016) 640–648.
[29] Z.L. Yang, X.F. Zeng, H.P. Liu, et al., Tetrahedron Lett. 57 (2016) 2579–2582.
[30] A. Geissner, C.L. Pereira, M. Leddermann, C. Anish, P.H. Seeberger, ACS Chem.
Biol. 11 (2016) 335–344.
This work was financially supported by the National Natural
Science Foundation of China (Nos. 21402140, 21502139), Shenzhen
Peacock Plan (No. KQTD2016053114253158), Natural Science
Foundation of Tianjin City (No. 16JCTPJC46000), Key Laboratory
of Industrial Fermentation Microbiology of Ministry of Education
and Tianjin Key Lab of Industrial Microbiology (No. 2015IM107)
and Youth Innovation Research Foundation of Tianjin University of
Science and Technology (No. 2016LG08). The authors are thankful
to the Research Centre of Modern Analytical Technology, Tianjin
University of Science and Technology for NMR and MALDI-TOF-MS
measurements. The authors also appreciate National Institute for
Viral Disease Control and Prevention, China CDC for the viral
strains.
[31] J.M. Katz, K. Hancock, X. Xu, Expert Rev. Anti. Infect. Ther. 9 (2011) 669–683.
[32] M. Mammen, G. Dahmann, G.M. Whitesides, J. Med. Chem. 38 (1995) 4179–
4190.
[33] S. Tang, W.B. Puryear, B.M. Seifried, et al., ACS Macro Lett. 5 (2016) 413–418.
[34] R. Bansil, B.S. Turner, Curr. Opin. Colloid Interface Sci. 11 (2006) 164–170.
[35] Y. He, Y. Yang, S.S. Iyer, Bioconjug. Chem. 27 (2016) 1509–1517.
[36] S. Barrett, P.G. Mohr, P.M. Schmidt, J.L. McKimm-Breschkin, PLoS ONE 6 (2011)
e23627.
[37] Y. Yang, Y. He, X. Li, H. Dinh, S.S. Iyer, Bioorg. Med. Chem. Lett. 24 (2014) 636–
643.
[38] S.J.F. Macdonald, K.G. Watson, R. Cameron, et al., Antimicrob. Agents.
Chemother. 48 (2004) 4542–4549.
References
[1] R. Schauer, Curr. Opin. Struct. Biol. 19 (2009) 507–514.
[2] C.W. Lloyd, Biol. Rev. Camb. Philos. Soc. 50 (1975) 325–350.
[3] N. Katopodis, Y. Hirshaut, N.L. Geller, C.C. Stock, Cancer Res. 42 (1982) 5270–
5275.
[4] J. Zhao, D.M. Simeone, D. Heidt, M.A. Anderson, D.M. Lubman, J. Proteome Res.
5 (2006) 1792–1802.
[39] S. Bhatia, L.C. Camacho, R. Haag, J. Am. Chem. Soc. 138 (2016) 8654–8666.
[40] V. Poonthiyil, P.T. Nagesh, M. Husain, V.B. Golovko, A.J. Fairbanks,
ChemistryOpen 4 (2015) 708–716.
[41] J.D. Driskell, C.A. Jones, S.M. Tompkins, R.A. Tripp, Analyst 136 (2011) 3083–
3090.
[5] S.S. Gokmen, C. Kazezoglu, B. Sunar, et al., Clin. Chem. Lab. Med. 44 (2006) 199–
206.
[6] G.R. Moe, T.S. Bhandari, B.A. Flitter, J. Immunol. 182 (2009) 6610–6617.
[7] S. Taube, M. Jiang, C.E. Wobus, Viruses 2 (2010) 1011–1049.
[8] S. Olofsson, T. Bergstrom, Ann. Med. 37 (2005) 154–172.
[9] C. Jones, M. Virji, P.R. Crocker, Mol. Microbiol. 49 (2003) 1213–1225.
[42] Z. Zhang, B. Schepens, L. Nuhn, et al., Chem. Commun. 52 (2016) 3352–3355.
[43] M. Yu, Y. Yang, R. Han, et al., Langmuir 26 (2010) 8534–8539.
Please cite this article in press as: X. Meng, et al., Multivalent neuraminidase hydrolysis resistant triazole-sialoside protein conjugates as
influenza-adsorbents, Chin. Chem. Lett. (2017), https://doi.org/10.1016/j.cclet.2017.10.032