Modification and Application of Glass Surfaces
Bioconjugate Chem., Vol. 21, No. 7, 2010 1247
small molecule-protein interactions. The results of these studies
Synthesis of 11-Chloroundecyldimethallylmethylsilane
(2b). A solution of H2PtCl6 · xH2O (10.4 mg, 0.02 mmol) in
2-propanol (0.2 mL) and dichloromethylsilane (5 mL, 48 mmol)
was added in a round-bottom flask, and the mixture was stirred
until it became homogeneous. Diethyl ether (10 mL) was added
into the reaction vessel, and the reaction temperature was raised
to 40 °C by an oil bath. After dropwise addition of 11-
chloroundec-1-ene (10 g, 52.98 mmol), the mixture was stirred
for 6 h at 80 °C. After the reaction, unreacted 11-chloroundec-
1-ene was removed by distillation. The remaining crude product,
dichloro-11-chloroundecylmethylsilane, was used in the next
step without further purification.
are described and discussed below.
MATERIALS AND METHODS
General. All chemical and biochemical products were of
analytical grade and purchased from commercial suppliers:
lectins from Vector Laboratories (Peterborough, U.K.) and
Sigma-Aldrich (St. Louis, MO); protein G from Streptococcus
sp. and goat anti-mouse FITC-labeled IgG from Sigma-Aldrich;
Cy3-streptavidin from Amersham Pharmacia biotech (Piscat-
away, NJ); ꢀ-1,4-galactosyltransferase (ꢀ-1,4-GalT) and UDP-
Gal from Calbiochem (Gibbstown, NJ); carbohydrates from
Sigma-Aldrich, Calbiochem (Gibbstown, NJ), TCI Corp. (To-
kyo, Japan), and Acros (Morris Plains, NJ); anti-dextran
antibody from Stemcell Technologies Inc. (Vancouver, BC,
Canada); microscope glass slide from Corning (Lowell, MA);
Dy547-RRE from Thermo Fisher Scientific (Lafayette, CO). The
solutions of probes were printed on the surface with a MicroSys
5100 from Cartesian Technologies Inc. (Irvine, CA). Microar-
rays probed with fluorescent dye labeled proteins and RNA were
scanned with an ArrayWoRx biochip reader from Applied
Precision (Northwest Issaquah, WA).
The crude dichloro-11-chloroundecylmethylsilane was added
dropwise into methallylmagnesium chloride (1.1 equiv) solution
in 300 mL of THF at 0 °C, and the reaction mixture was stirred
for 6 h at 0 °C. After the reaction, saturated aqueous NH4Cl
was added to the reaction mixture, and the mixture was extracted
with diethyl ether three times. The collected organic layers were
dried over anhydrous MgSO4 and filtered through Celite pad.
The crude mixture was purified by flash column chromatography
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(n-hexane; Rf ) 0.8) to give 2b in 65% yield (10.71 g): H
NMR (250 MHz, CDCl3) δ 4.74 (d, J ) 26.6 Hz, 4 H), 3.70 (t,
J ) 6.74 Hz, 2 H), 2.04-1.91 (m, 8 H), 1.64-1.48 (m, 18 H),
0.77-0.74 (m, 2 H), 0.25 (s, 3 H); 13C NMR (62.9 MHz, CDCl3)
δ 143.4, 108.9, 45.0, 33.9, 32.9, 29.7, 29.6, 29.4, 29.1, 27.1,
26.3, 25.9, 25.5, 23.9, 14.3, 4.2; IR spectrum (neat) 3073, 2924,
2853, 1636, 1448, 1279, 1250, 870 cm-1. Anal. Calcd for
C20H39ClSi: C, 70.02; H, 11.46. Found: C, 70.08; H, 11.79.
Synthesis of 3-Chloropropyldimethallylmethylsilane (2a). A
solution of H2PtCl6 ·xH2O (10.4 mg, 0.02 mmol) in 2-propanol
(0.2 mL) and dichloromethylsilane (26 mL, 250 mmol) was
added in a round-bottom flask, and the mixture was stirred until
it became homogeneous. Diethyl ether (10 mL) was added into
the reaction vessel, and the reaction temperature was raised to
40 °C by an oil bath. After dropwise addition of allyl chloride
(20 mL, 250 mmol), the mixture was stirred for 6 h at 80 °C.
After the reaction, all volatiles were evaporated under the
reduced pressure. The crude product, 3-chloropropyldichlorom-
ethylsilane, was used in the next step without further purification.
Synthesis of 11-Azidoundecyldimethallylmethylsilane (3b). 11-
Chloroundecyldimethallylsilane (2b, 5.5 g, 16.7 mmol) and
sodium azide (2.18 g, 33.5 mmol) were added to 100 mL of
DMF and stirred for 4 h at 80 °C. After the reaction, saturated
aqueous NH4Cl solution was added. The solution was extracted
with diethyl ether three times. The collected organic layers were
dried over anhydrous MgSO4 and filtered through Celite pad.
The crude mixture was purified by flash column chromatography
(n-hexane/ethyl acetate ) 10:1, Rf ) 0.6) to give 3b in 78%
yield (4.56 g): 1H NMR (250 MHz, CDCl3) δ 4.51 (d, J ) 27.3
Hz, 4 H), 3.22 (t, J ) 6.9 Hz, 2 H), 1.68 (s, 6 H), 1.58 (m, 6
The crude 3-chloropropyldichloromethylsilane was added
dropwise into methallylmagnesium chloride (1.1 equiv) solution
in 300 mL of THF at 0 °C, and the reaction mixture was stirred
for 6 h at 0 °C. After the reaction, saturated aqueous NH4Cl
was added to the reaction mixture, and the mixture was extracted
with diethyl ether three times. The collected organic layers were
dried over anhydrous MgSO4 and filtered through Celite pad.
The crude mixture was purified by flash column chromatography
H), 1.24 (s, 18 H), 0.54 (t, J ) 7.8 Hz, 2 H), 0.01 (s, 3 H); 13
C
NMR (62.9 MHz, CDCl3) δ 143.7, 108.9, 51.7, 33.9, 30.0, 29.8,
29.5, 29.4, 29.1, 26.9, 25.9, 25.6, 23.9, 14.3, -4.3; IR spectrum
(neat) 3072, 2921, 2851, 2094, 1636, 1454, 1373, 1279, 1250,
1166, 998, 971, 868, 839, 722 cm-1. Anal. Calcd for C20H39N3Si:
C, 68.71; H, 11.24; N, 12.02. Found: C, 68.60; H, 12.15; N,
12.15.
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(n-hexane; Rf ) 0.6) to give 2a in 70% yield (40 g): H NMR
(250 MHz, CDCl3) δ 4.57 (d, J ) 30.7 Hz, 4 H), 3.50 (t, J )
13.9 Hz, 2 H), 1.86-1.76 (m, 2 H), 1.73 (s, 6 H), 1.60 (s, 4 H),
0.73-0.68 (m, 2 H), 0.09 (s, 3 H); 13C NMR (62.9 MHz, CDCl3)
δ 143.3, 109.3, 48.1, 27.7, 25.8, 25.6, 11.9, 4.2; IR spectrum
(neat) 3083, 2966, 2914, 1644, 1450, 1748, 1378, 1280, 1168,
871, 840 cm-1. Anal. Calcd for C12H23ClSi: C, 62.43; H, 10.04.
Found: C, 62.48; H, 9.93.
Synthesis of 2,5-Dioxopyrrolidin-1-yl-1-(3-(methylbis(2-me-
thylallyl)silyl)propyl)-1H-1,2,3-triazole 4-Carboxylate (1a). A
solution of propiolic acid (2.1 mL, 34.0 mmol), N-hydroxysuc-
cinimide (4.0 g, 34.8 mmol), and N,N′-dicyclohexylcarbodiimide
(8.6 g, 41.7 mmol) in 80 mL of dimethoxyethane was stirred
for 18 h at room temperature. After the reaction, the mixture
was filtered through Celite pad to remove dicyclohexylurea, and
all volatiles were evaporated under the reduced pressure. The
crude product, 2,5-dioxopyrrolidin-1-yl propiolate, was used in
the next step without further purification. To a solution of crude
2,5-dioxopyrrolidin-1-yl propiolate in 50 mL of THF was added
CuSO4 ·5H2O (0.43 g, 1.74 mmol), sodium ascorbate (0.69 g,
3.48 mmol) in 50 mL of H2O, and 3-azidopropylmeth-
yldimethallylsilane (3a, 4.13 g, 17.38 mmol). The reaction
mixture was stirred for 4 h at room temperature. After the
reaction, saturated aqueous NH4Cl was added, and the resulting
mixture was extracted with diethyl ether three times. The
collected organic layers were dried over anhydrous MgSO4 and
filtered through Celite pad, then purified by flash column
chromatography (n-hexane/ethyl acetate ) 1:1, Rf ) 0.3) to give
Synthesis of 3-Azidopropyldimethallylmethylsilane (3a).
3-Chloropropyldimethallylmethylsilane (2a, 10 g, 43.2 mmol)
and sodium azide (5.63 g, 86.58 mmol) was added to 100 mL
of DMF and stirred for 4 h at 80 °C. After the reaction, saturated
aqueous NH4Cl solution was added. The solution was extracted
with diethyl ether three times. The collected organic layers were
dried over anhydrous MgSO4 and filtered through Celite pad.
The crude mixture was purified by flash column chromatography
(n-hexane/ethyl acetate ) 10:1, Rf ) 0.6) to give 3a in 92%
yield (9.5 g): 1H NMR (250 MHz, CDCl3) δ 4.57 (d, J ) 28.5
Hz, 4 H), 3.23 (t, J ) 7.0 Hz, 2 H), 1.78 (s, 6 H), 1.77-1.60
(m, 2 H), 1.58 (s, 4 H), 0.66-0.60 (m, 2 H), 0.07 (s, 3 H); 13
C
NMR (62.9 MHz, CDCl3) δ 143.3, 109.3, 54.7, 25.8, 25.6, 23.8,
11.4, 4.2; IR spectrum (neat) 3079, 2971, 2922, 2876, 2102,
1748, 1636, 1459, 1378, 1282, 1255, 1170, 873, 846 cm-1. Anal.
Calcd for C12H23N3Si: C, 60.71; H, 9.76; N, 17.70. Found: C,
60.68; H, 9.72; N, 16.78.
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1a in 60% yield (4.21 g): H NMR (250 MHz, CDCl3) δ 8.33