ペプチド・タンパク質のための分析・分取用カラム カタログ...
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PowerPoint T009J
10 mg
C182002
HC18-2150 LCMS 2.0 id X 150 0.150.2 mL/min
HC18-3150 3.0 id X 150 0.5 mL/min
HC18-46150 4.6 id X 150 1.0 mL/min
HC18-20150 20 id X 150 1020 mL/min
HC18-10250 10 id X 250 23 mL/min
HC18-20250 20 id X 250 1020 mL/min
HC18-28250 28 id X 250 2040 mL/min
HiPep-Cadenza C18
0.45 μm0.2 μm
20 MPa
pHpH 27pH
pH
HiPep-Cadenza C18
HiPep-Cadenza C18
HiPep-Cadenza C18C18
HiPep-Cadenza C184.6 id X 150 mm3.0 id X 150 mmC18
4.6 id X 150 mm
Arg 4Lys 9 3200
A=0.01N HCl B=0.01N HCl in 90% Acetonitrile A/B=95/15-65/35 in 30min UV 210 nm
min 0 2 4 6 8 10 12 14 16 18 20 22 24 26 28 30
m A
B S
150 A (210nm) TO6016-586H-TFA221002 40224AC05
min 0 2 4 6 8 10 12 14 16 18 20 22 24 26 28 30
m A
B S
A (210nm) TO6016-586HTFA221002 030304C 40308AC02
min 0 2 4 6 8 10 12 14 16 18 20 22 24 26 28 30
m A
B S
120 A (210nm) TO6016-586H-TFA221002 40223AC03
HiPep-Cadenza C18 3.0 id X 150 mm : 0.5 mL/min (1% = 1 min)
C18 4.6 id X 150 mm : 1.0 mL/min (1% = 1 min)
3
T009J
: 1.0 mL/min (1% = 1 min)
HiPep-Intrada
HiPep-Intrada
HiPep-Cadenza C18HiPep-
Cadenza C18HiPep-Intrada HiPep-Cadenza C18
HiPep-IntradaHiPep-Cadenza C18
HiPep-Intrada3.0 id ×150 mm 0.3 mL/minUV 210nm
A= 0.1% TFA, B= 0.1% TFA in 90% AcetonitrileGradient A/B= 55/45-25/75% in 30 min at 60
HiPep-Intrada 20 id x 250 mm60
15
*LCMSMS/MS
HiPep-Intrada
Gradient1000
100%
30%0.1%
HiWR-2150 LCMS* 2.0 id X 150 0.150.2 mL/min
HiWR-3150 3.0 id X 150 0.50.75 mL/min
HiWR-46150 4.6 id X 150 0.751.25 mL/min
HiWR-10150 10 id X 150 23 mL/min
HiWR-10250 10 id X 250 23 mL/min
HiWR-20250 20 id X 250 1020 mL/min
HiWR-28250 28 id X 250 2040 mL/min
4
T009J
βAmyloid (Aβ) HiPep- IntradaC18
Hirata, A., Miyajima, M., and Nokihara, K. Separation of peptides having structures and derivatives of mimosine, a non- proteinogenic amino acid, by a novel reverse-phase HPLC column packed with wide-pore silica, J. Liquid Chromatography & Related Technologies, 36, 2960-2967, 2013.
HiPep-Cadenza C18HiPep-IntradaAβ
HiPep-Intrada ODS3.0 id x 150 mmβ1-43 4615.10
0.0 5.0 10.0 15.0 min
0.0
2.5
5.0
0
50
100
150
0.0 5.0 10.0 15.0 20.0 25.0 min
0
50
100
150
200
Mass (m/z)
4616.1269
4617.0778
4618.0602
4615.0981
4619.07294614.0431
4620.0979
5
T009J
PrP (a), (b); PrP(106-126), (c), (d); PrP[(169-176)+180-195)]
(a), (c); HiPep-Cadenza C18, (b), (d); HiPep-Intrada 60
HiPep-Intrada
(a), (b); GITC-D,L-Ala (c), (d); GITC-D,L-Mimosine
(a), (c); C18 (b), (d); HiPep-Intrada
GITC
HiPep-Cadenza C18HiPep-Intrada
6
T009J
100 120 210 μm
ODS
FDA Taspogluchide (GLP-1)id 20 mmTaspogluchide ODS Daisogel® SP65100 Daisogel® SP
2010EPS Nokihara, K., et al., Peptides, Proceedings of the 31st European Peptide Symposium, Lebl, M., Meldal, M., and Jensen, K. J.,, Hoeg-Jensen, T. (Editors), pp136-137, 2010.
Daisogel® SP
LC
: HiPep-Intrada20 id x 150 mm60 ºC, : A; 0.1% TFA in H2O, B; 0.1% TFA in 90% Acetonitrile, : 9.9 mL/min, : UV at 210 nm
1. A
DMSO
min 0 2 4 6 8 10 12 14 16 18 20 22 24 26 28 30
m AB
<70%
min 0 2 4 6 8 10 12 14 16 18 20 22 24 26 28 30
m AB
2500 A (215nm) HPP-h12-TFA100909 N90916C07
0.0 2.5 5.0 7.5 10.0 12.5 15.0 17.5 20.0 22.5 25.0 27.5 min 0
25
50
75
100
125
150
175
200
225
250
275
mV A:210nm
>90%
min 0.0 2.5 5.0 7.5 10.0 12.5 15.0 17.5 20.0 22.5 25.0 27.5 30.0 32.5 35.0 37.5 40.0
m AB
DMSO
min 0.0 2.5 5.0 7.5 10.0 12.5 15.0 17.5 20.0 22.5 25.0 27.5 30.0 32.5 35.0 37.5 40.0
m AB
>90%
3. C DMSO HFIP
min 0.0 2.5 5.0 7.5 10.0 12.5 15.0 17.5 20.0 22.5 25.0 27.5 30.0 32.5 35.0 37.5 40.0
m AB
2500 A (215nm) HPP-h23-TFA090909 N90917C02
0.0 2.5 5.0 7.5 10.0 12.5 15.0 17.5 20.0 22.5 25.0 27.5 min 0
25
50
75
100
125
150
175
200mVA:210nm
>90%
min 0.0 2.5 5.0 7.5 10.0 12.5 15.0 17.5 20.0 22.5 25.0 27.5 30.0 32.5 35.0 37.5 40.0
m AB
2500 A (215nm) HPP-h23-TFA090909 N90917C01
0.0 2.5 5.0 7.5 10.0 12.5 15.0 17.5 20.0 22.5 25.0 27.5 min
0
10
20
30
40
50
60
70
80
90
100mVA:210nm
602-8158 486-46 TEL: 075-813-2101, FAX: 075-801-0280, E-mail: [email protected], URL: http://www.hipep.jp/
1. HiPep-Cadeza ODS Ab : 0.5 mL/min, : 210 nm, : A = 0.1% TFA, B = 0.1% TFA, 90% (a) Aβ (21-42); 2070.9 [M+H]+, (b) Aβ (1-40); 2165.4 [M+2H] 2+, (c) Aβ (17-40); 2393.1 [M+H]+, (d) Aβ (1-20) 2461.7 [M+H]+.
b-
602-8158 486-46
(AD) [1] AD [2] 39-43 [3]AβAD43 Aβ (1-43)ODSAβ (1-43) Zorbax® HPLC80NH4OH AβAβ Aβ
AβAβ (1-43) ; Mwt 4615.21Aβ (1-42) ; Mwt 4514.10 Aβ (17-42) ; Mwt 2577.08Aβ (21-42) ; mwt 2070.44Aβ (1-40) ; mwt 4329.86Aβ (17-40) ; mwt 2392.85Aβ (1-20) ; mwt 2461.687 PSSM-8TentaGel® ResinFmocFmoc 2% 2% DBU DMFPyBOP / HOBt / NMM10[4] ‘1 ’ [5]TFA / H2O / Anisol / EDT / EMS / Phenol = 82/5/5/3/2/3 (v/v %) 90HiPep-Cadenza ODS3.0 id × 150 mm LC-MS Aβ (21-42)Aβ (1-40) Aβ (17-40) Aβ (1-20) 1Aβ (1-43)Aβ (1- 42)Aβ (17-42)HiPep-Cadenza ODSHPLC 2eNMR Aβ (1-40)CAβ (1-42)C [6]
2. Aβ (1-43) HiPep-Cadenza ODS (e) HiPep-Intrada (f) : 0.5 mL/min, :210 nm, : A = 0.1% TFA, B = 0.1% TFA, 90% (e) 18-30 2308.3 [M+2H]2+ . : . (b) 2308.3 [M+2H]2+ . : 60.
44 200710 1/2
0 10 20 min
0
250
500
750
0
50
100
150
200
4 5 hPrP(90-142) HiPep- Cadenza 5aHiPep-Intrada 5b 6.AβCD Aβ (1-40) Aβ (1-42)Aβ (1-43)β-Aβ (1-42)Aβ (1-43)
Aβpseudoproline HATU HiPep-Intrada ( 3 mm 5 mm) PepTenChip®
7
T009J
0 10 20 min
a b
0.0 2.5 5.0 7.5 10.0 12.5 15.0 17.5 20.0 22.5 25.0 27.5 min
-50
-25
0
25
50
75
100
125
150
175 mV A:210nm
0.0 2.5 5.0 7.5 10.0 12.5 15.0 17.5 20.0 22.5 25.0 27.5 min
0
10
20
30
40
50
60
70
80
ba c
3. HiPep-Intrada Aβ (1-42) Aβ (17-42) ( LC- MS, : 0.5 mL/min, : 210 nm, : A = 0.1% TFA, B = 0.1% TFA, 90% , : 60). (a) Aβ (1-42); 2257.3 [M+2H] 2+, (b) Aβ (17-42); 2576.9 [M+H]+.
5. hPrP(90-142) . (a) ODS (HiPep-Cadenza). (b) HiPep-Intrada .
6. CD Aβ . (a) Aβ(1-40), (b) Aβ(1-42), (c) Aβ(1-43), 0.1 mM 10 mM (pH 7.0)
1. Newman, M., Musgrave, I., and Lardeli, M. (2007) Biochemica et Biophysica Acta, 1772, 285-297. 2. Rudolph, E. Tanzi. and Lars Bertram. (2005) Cell, 120, 545-555. 3. Glenner, G. G. and Wong, C. W. (1984) Biochem. Biophys. Res. Commun., 122, 1131-1135. 4. Nokihara, K., Yasuhara, T., Nakata, Y., Lerner, E. A. and Wray, V. (2007) Int. J. Peptide Res. Therapeutics, 13, 377-386. 5. Nokihara, K., Yamaguchi, M., Ando, E., Kuriki, T., and Yokomizo, Y. (1996) In Peptide Chemistry 1995, Nishi, N. (ed) Protein Research
Foundation Osaka, 65-68. 6. Lim, K. H., Collver, H. H., Le. Y. T. H., Nagchowdhuri, P. and Kenny, J. M. (2007) Biochem. Biophys. Res. Commun., 353, 443-449.
44 200710 2/2
4. HiPep-Intrada Aβ (1-43)MALDI-TOF-MS
14-18 min
10 mg
C182002
HC18-2150 LCMS 2.0 id X 150 0.150.2 mL/min
HC18-3150 3.0 id X 150 0.5 mL/min
HC18-46150 4.6 id X 150 1.0 mL/min
HC18-20150 20 id X 150 1020 mL/min
HC18-10250 10 id X 250 23 mL/min
HC18-20250 20 id X 250 1020 mL/min
HC18-28250 28 id X 250 2040 mL/min
HiPep-Cadenza C18
0.45 μm0.2 μm
20 MPa
pHpH 27pH
pH
HiPep-Cadenza C18
HiPep-Cadenza C18
HiPep-Cadenza C18C18
HiPep-Cadenza C184.6 id X 150 mm3.0 id X 150 mmC18
4.6 id X 150 mm
Arg 4Lys 9 3200
A=0.01N HCl B=0.01N HCl in 90% Acetonitrile A/B=95/15-65/35 in 30min UV 210 nm
min 0 2 4 6 8 10 12 14 16 18 20 22 24 26 28 30
m A
B S
150 A (210nm) TO6016-586H-TFA221002 40224AC05
min 0 2 4 6 8 10 12 14 16 18 20 22 24 26 28 30
m A
B S
A (210nm) TO6016-586HTFA221002 030304C 40308AC02
min 0 2 4 6 8 10 12 14 16 18 20 22 24 26 28 30
m A
B S
120 A (210nm) TO6016-586H-TFA221002 40223AC03
HiPep-Cadenza C18 3.0 id X 150 mm : 0.5 mL/min (1% = 1 min)
C18 4.6 id X 150 mm : 1.0 mL/min (1% = 1 min)
3
T009J
: 1.0 mL/min (1% = 1 min)
HiPep-Intrada
HiPep-Intrada
HiPep-Cadenza C18HiPep-
Cadenza C18HiPep-Intrada HiPep-Cadenza C18
HiPep-IntradaHiPep-Cadenza C18
HiPep-Intrada3.0 id ×150 mm 0.3 mL/minUV 210nm
A= 0.1% TFA, B= 0.1% TFA in 90% AcetonitrileGradient A/B= 55/45-25/75% in 30 min at 60
HiPep-Intrada 20 id x 250 mm60
15
*LCMSMS/MS
HiPep-Intrada
Gradient1000
100%
30%0.1%
HiWR-2150 LCMS* 2.0 id X 150 0.150.2 mL/min
HiWR-3150 3.0 id X 150 0.50.75 mL/min
HiWR-46150 4.6 id X 150 0.751.25 mL/min
HiWR-10150 10 id X 150 23 mL/min
HiWR-10250 10 id X 250 23 mL/min
HiWR-20250 20 id X 250 1020 mL/min
HiWR-28250 28 id X 250 2040 mL/min
4
T009J
βAmyloid (Aβ) HiPep- IntradaC18
Hirata, A., Miyajima, M., and Nokihara, K. Separation of peptides having structures and derivatives of mimosine, a non- proteinogenic amino acid, by a novel reverse-phase HPLC column packed with wide-pore silica, J. Liquid Chromatography & Related Technologies, 36, 2960-2967, 2013.
HiPep-Cadenza C18HiPep-IntradaAβ
HiPep-Intrada ODS3.0 id x 150 mmβ1-43 4615.10
0.0 5.0 10.0 15.0 min
0.0
2.5
5.0
0
50
100
150
0.0 5.0 10.0 15.0 20.0 25.0 min
0
50
100
150
200
Mass (m/z)
4616.1269
4617.0778
4618.0602
4615.0981
4619.07294614.0431
4620.0979
5
T009J
PrP (a), (b); PrP(106-126), (c), (d); PrP[(169-176)+180-195)]
(a), (c); HiPep-Cadenza C18, (b), (d); HiPep-Intrada 60
HiPep-Intrada
(a), (b); GITC-D,L-Ala (c), (d); GITC-D,L-Mimosine
(a), (c); C18 (b), (d); HiPep-Intrada
GITC
HiPep-Cadenza C18HiPep-Intrada
6
T009J
100 120 210 μm
ODS
FDA Taspogluchide (GLP-1)id 20 mmTaspogluchide ODS Daisogel® SP65100 Daisogel® SP
2010EPS Nokihara, K., et al., Peptides, Proceedings of the 31st European Peptide Symposium, Lebl, M., Meldal, M., and Jensen, K. J.,, Hoeg-Jensen, T. (Editors), pp136-137, 2010.
Daisogel® SP
LC
: HiPep-Intrada20 id x 150 mm60 ºC, : A; 0.1% TFA in H2O, B; 0.1% TFA in 90% Acetonitrile, : 9.9 mL/min, : UV at 210 nm
1. A
DMSO
min 0 2 4 6 8 10 12 14 16 18 20 22 24 26 28 30
m AB
<70%
min 0 2 4 6 8 10 12 14 16 18 20 22 24 26 28 30
m AB
2500 A (215nm) HPP-h12-TFA100909 N90916C07
0.0 2.5 5.0 7.5 10.0 12.5 15.0 17.5 20.0 22.5 25.0 27.5 min 0
25
50
75
100
125
150
175
200
225
250
275
mV A:210nm
>90%
min 0.0 2.5 5.0 7.5 10.0 12.5 15.0 17.5 20.0 22.5 25.0 27.5 30.0 32.5 35.0 37.5 40.0
m AB
DMSO
min 0.0 2.5 5.0 7.5 10.0 12.5 15.0 17.5 20.0 22.5 25.0 27.5 30.0 32.5 35.0 37.5 40.0
m AB
>90%
3. C DMSO HFIP
min 0.0 2.5 5.0 7.5 10.0 12.5 15.0 17.5 20.0 22.5 25.0 27.5 30.0 32.5 35.0 37.5 40.0
m AB
2500 A (215nm) HPP-h23-TFA090909 N90917C02
0.0 2.5 5.0 7.5 10.0 12.5 15.0 17.5 20.0 22.5 25.0 27.5 min 0
25
50
75
100
125
150
175
200mVA:210nm
>90%
min 0.0 2.5 5.0 7.5 10.0 12.5 15.0 17.5 20.0 22.5 25.0 27.5 30.0 32.5 35.0 37.5 40.0
m AB
2500 A (215nm) HPP-h23-TFA090909 N90917C01
0.0 2.5 5.0 7.5 10.0 12.5 15.0 17.5 20.0 22.5 25.0 27.5 min
0
10
20
30
40
50
60
70
80
90
100mVA:210nm
602-8158 486-46 TEL: 075-813-2101, FAX: 075-801-0280, E-mail: [email protected], URL: http://www.hipep.jp/
1. HiPep-Cadeza ODS Ab : 0.5 mL/min, : 210 nm, : A = 0.1% TFA, B = 0.1% TFA, 90% (a) Aβ (21-42); 2070.9 [M+H]+, (b) Aβ (1-40); 2165.4 [M+2H] 2+, (c) Aβ (17-40); 2393.1 [M+H]+, (d) Aβ (1-20) 2461.7 [M+H]+.
b-
602-8158 486-46
(AD) [1] AD [2] 39-43 [3]AβAD43 Aβ (1-43)ODSAβ (1-43) Zorbax® HPLC80NH4OH AβAβ Aβ
AβAβ (1-43) ; Mwt 4615.21Aβ (1-42) ; Mwt 4514.10 Aβ (17-42) ; Mwt 2577.08Aβ (21-42) ; mwt 2070.44Aβ (1-40) ; mwt 4329.86Aβ (17-40) ; mwt 2392.85Aβ (1-20) ; mwt 2461.687 PSSM-8TentaGel® ResinFmocFmoc 2% 2% DBU DMFPyBOP / HOBt / NMM10[4] ‘1 ’ [5]TFA / H2O / Anisol / EDT / EMS / Phenol = 82/5/5/3/2/3 (v/v %) 90HiPep-Cadenza ODS3.0 id × 150 mm LC-MS Aβ (21-42)Aβ (1-40) Aβ (17-40) Aβ (1-20) 1Aβ (1-43)Aβ (1- 42)Aβ (17-42)HiPep-Cadenza ODSHPLC 2eNMR Aβ (1-40)CAβ (1-42)C [6]
2. Aβ (1-43) HiPep-Cadenza ODS (e) HiPep-Intrada (f) : 0.5 mL/min, :210 nm, : A = 0.1% TFA, B = 0.1% TFA, 90% (e) 18-30 2308.3 [M+2H]2+ . : . (b) 2308.3 [M+2H]2+ . : 60.
44 200710 1/2
0 10 20 min
0
250
500
750
0
50
100
150
200
4 5 hPrP(90-142) HiPep- Cadenza 5aHiPep-Intrada 5b 6.AβCD Aβ (1-40) Aβ (1-42)Aβ (1-43)β-Aβ (1-42)Aβ (1-43)
Aβpseudoproline HATU HiPep-Intrada ( 3 mm 5 mm) PepTenChip®
7
T009J
0 10 20 min
a b
0.0 2.5 5.0 7.5 10.0 12.5 15.0 17.5 20.0 22.5 25.0 27.5 min
-50
-25
0
25
50
75
100
125
150
175 mV A:210nm
0.0 2.5 5.0 7.5 10.0 12.5 15.0 17.5 20.0 22.5 25.0 27.5 min
0
10
20
30
40
50
60
70
80
ba c
3. HiPep-Intrada Aβ (1-42) Aβ (17-42) ( LC- MS, : 0.5 mL/min, : 210 nm, : A = 0.1% TFA, B = 0.1% TFA, 90% , : 60). (a) Aβ (1-42); 2257.3 [M+2H] 2+, (b) Aβ (17-42); 2576.9 [M+H]+.
5. hPrP(90-142) . (a) ODS (HiPep-Cadenza). (b) HiPep-Intrada .
6. CD Aβ . (a) Aβ(1-40), (b) Aβ(1-42), (c) Aβ(1-43), 0.1 mM 10 mM (pH 7.0)
1. Newman, M., Musgrave, I., and Lardeli, M. (2007) Biochemica et Biophysica Acta, 1772, 285-297. 2. Rudolph, E. Tanzi. and Lars Bertram. (2005) Cell, 120, 545-555. 3. Glenner, G. G. and Wong, C. W. (1984) Biochem. Biophys. Res. Commun., 122, 1131-1135. 4. Nokihara, K., Yasuhara, T., Nakata, Y., Lerner, E. A. and Wray, V. (2007) Int. J. Peptide Res. Therapeutics, 13, 377-386. 5. Nokihara, K., Yamaguchi, M., Ando, E., Kuriki, T., and Yokomizo, Y. (1996) In Peptide Chemistry 1995, Nishi, N. (ed) Protein Research
Foundation Osaka, 65-68. 6. Lim, K. H., Collver, H. H., Le. Y. T. H., Nagchowdhuri, P. and Kenny, J. M. (2007) Biochem. Biophys. Res. Commun., 353, 443-449.
44 200710 2/2
4. HiPep-Intrada Aβ (1-43)MALDI-TOF-MS
14-18 min