US2017067904A1PendingUtilityA1
Compositions and methods for membrane protein detergent stability screen
Assignee: UNIV VIRGINIA PATENT FOUNDATIONPriority: Jun 29, 2009Filed: Nov 18, 2016Published: Mar 9, 2017
Est. expiryJun 29, 2029(~2.9 yrs left)· nominal 20-yr term from priority
B01D 15/3804C30B 7/08G01N 33/6803
47
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Claims
Abstract
The invention provides methods to assess protein stability and to obtain sizing information. In one aspect, the screen comprises a 94 detergent panel and a series of MWCO filtered microplates. A protein of interest is bound to an affinity matrix and aliquoted into a 96-well microplate. Wells containing the immobilized protein are washed in the new detergent and then eluted in the new detergent into a collection plate. Protein not stable in the new detergent is precipitated on the resin and not present in the elutions. Half of the elution is passed through a high (i.e., 300 kDa) MWCO microplate and the other half through a low (i.e., 100 kDa) MWCO microplate. Elutions from the microplates are spotted on a nitrocellulose membrane, visualized by Western analysis (or by some other method), and quantified. The high MWCO provides stability readout and the ratio of low/high kDa provides sizing information.
Claims
exact text as granted — not AI-modifiedWhat is claimed is:
1 . A method for determining the stability and size of a protein in a detergent, said method comprising:
a) obtaining a solution comprising said protein in a first detergent; b) adding an effective amount of an affinity resin to said solution; c) adding an aliquot of said solution comprising said protein in a first detergent and an affinity resin to a first chamber, said first chamber comprising a filter wherein said pore size is about 0.2 μm; d) washing said first chamber with a wash solution comprising a different detergent; e) eluting said protein in said first chamber with an elution solution comprising said different detergent and collecting the eluate; f) passing an aliquot of said eluate through a second chamber comprising a high molecular weight cut-off filter and another aliquot of said eluate through a third chamber comprising a low molecular weight cut-off filter; g) measuring the amount of protein in the eluate passing through the high molecular weight cut-off filter and the amount of protein in the eluate passing through the low molecular weight cut-off filter; and h) comparing the amount of protein eluted through the high molecular weight cut-off filter with the amount of protein eluted through the low molecular weight cut-off filter, thereby determining the stability and size of a protein in at least one detergent.
2 . The method of claim 1 , wherein said first chamber is a well of a multiwell microplate.
3 . The method of claim 1 , wherein said second chamber is a well of a multiwell microplate, further wherein each well comprises a high molecular weight cut-off filter.
4 . The method of claim 1 , wherein said third chamber is a well of a multiwell microplate, further wherein each well comprises a low molecular weight cut-off filter.
5 . The method of claim 2 , wherein at least two wells are used.
6 . The method of claim 5 , wherein multiple different detergents are used.
7 . The method of claim 6 , wherein when multiple detergents are tested, each well comprising a detergent comprises a different detergent than the other wells comprising a detergent, and optionally one or more wells comprise a positive control and one or more wells comprise a negative control.
8 . The method of claim 1 , wherein about 20 column volumes of said wash solution are used for said wash.
9 . The method of claim 1 , wherein about 6 column volumes of said elution solution are used for said elution.
10 . The method of claim 1 , wherein said high molecular weight cut-off filter is about 300 kDa.
11 . The method of claim 1 , wherein said low molecular weight cut-off filter is about 100 kDa.
12 . The method of claim 7 , wherein said multiwell plate is a 96, 384, or 1536 well plate.
13 . The method of claim 12 , wherein said 96 well plate is a Society for Biomolecular Sciences format plate.
14 . The method of claim 12 , wherein 94 different detergents are tested.
15 . The method of claim 1 , wherein said different detergent is selected from the following table:
Well
Abbrev.
Name
[Det] mM
A1
No detergent (−control)
A2
Empty well for current detergent (+control)
A3
Z3-12
ZWITTERGENT ® 3-12
8.4
(2.8)
A4
Z3-14
ZWITTERGENT ® 3-14
10
(0.2)
A5
DMG
n-Decyl-N,N-dimethylglycine
38
(19)
A6
DOMG
n-Dodecyl-N,N-dimethylglycine
4.5
(1.5)
A7
DAO
n-Decyl-N,N-dimethylamine-N-oxide
21
(10.5)
A8
UDAO
n-Undecyl-N,N,-dimethylamine-N-oxide
9.6
(3.2)
A9
LDAO
n-Dodecyl-N,N-dimethylamine-N-oxide
3
(1)
A10
C-DDFOS
C-DODECAFOS ™
44
(22)
A11
CF-4
CYCLOFOS ™-4
28
(14)
A12
CF-5
CYCLOFOS ™-5
13.5
(4.5)
B1
CF-6
CYCLOFOS ™-6
8.04
(2.68)
B2
CF-7
CYCLOFOS ™-7
6.2
(0.62)
B3
FC-10
FOS-CHOLINE ®-10
22
(11)
B4
FC-11
FOS-CHOLINE ®-11
5.55
(1.85)
B5
FC-12
FOS-CHOLINE ®-12
4.5
(1.5)
B6
FC-13
FOS-CHOLINE ®-13
7.5
(0.75)
B7
FC-14
FOS-CHOLINE ®-14
6
(0.12)
B8
FC-I11
FOS-CHOLINE ®-ISO-11
53.2
(26.6)
B9
FC-I11-6U
FOS-CHOLINE ®-ISO-11-6U
51.6
(25.8)
B10
FC-I9
FOS-CHOLINE ®-ISO-9
64
(32)
B11
FC-U10-11
FOS-CHOLINE ®-UNSAT-11-10
15.5
(6.2)
B12
DHPC
1,2-Diheptanoyl-sn-glycero-3-phosphocholine
4.2
(1.4)
C1
LPC-10
LysoPC-10
20
(8)
C2
LPC-12
LysoPC-12
7
(0.7)
C3
FOSFEN-9
FOSFEN ™-9
4.05
(1.35)
C4
CHAPS
CHAPS
20
(8)
C5
CHAPSO
CHAPSO
20
(8)
C6
DDMAU
n-Dodecyl-N,N-(dimethylammonio)undecanoate
6.5
(0.13)
C7
DDMAB
n-Dodecyl-N,N-(dimethylammonio)butyrate
12.9
(4.3)
C8
LAPAO
LAPAO
4.8
(1.6)
C9
TRIPAO
TRIPAO
13.5
(4.5)
C10
T-20
TWEEN ® 20
5.9
(0.059)
C11
BRIJ-35
BRIJ ®35
9.1
(0.091)
C12
TX-100
TRITON ® X-100
11.5
(0.23)
D1
TX-114
TRITON ® X-114
10
(0.2)
D2
TX-305
TRITON ® X-305
6.5
(0.65)
D3
TX-405
TRITON ® X-405
8.1
(0.81)
D4
NID-P40
[Octylphenoxy]polyethoxyethanol
15
(0.3)
D5
APO8
Dimethyloctylphosphine oxide
80
(40)
D6
APO9
Dimethylnonylphosphine oxide
20
(10)
D7
APO10
Dimethyldecylphosphine oxide
13.98
(4.66)
D8
APO11
Dimethylundecylphosphine oxide
3.6
(1.2)
D9
APO12
Dimethyldodecylphosphine oxide
5.7
(0.57)
D10
C6E3
Triethylene glycol monohexyl ether
46
(23)
D11
C6E4
Tetraethylene glycol monohexyl ether
60
(30)
D12
C6E5
Pentaethylene glycol monohexyl ether
74
(37)
E1
C7E5
Pentaethylene glycol monoheptyl ether
42
(21)
E2
C8E4
Tetraethylene glycol monooctyl ether
20
(8)
E3
C8E5
Pentaethylene glycol monooctyl ether
17.75
(7.1)
E4
C8E6
Hexaethylene glycol monooctyl ether
25
(10)
E5
C10E5
Pentaethylene glycol monodecyl ether
8.1
(0.81)
E6
C10E6
Hexaethylene glycol monodecyl ether
9
(0.9)
E7
C10E9
Polyoxyethylene(9)decyl ether
3.9
(1.3)
E8
C12E8
Octaethylene glycol monododecyl ether
9
(0.09)
E9
C12E9
Polyoxyethylene(9)dodecyl ether
5
(0.05)
E10
C12E10
Polyoxyethylene(10)dodecyl ether
10
(0.2)
E11
C13E8
Polyoxyethylene(8)tridecyl ether
10
(0.1)
E12
CHAP
Big CHAP
8.7
(2.9)
F1
CHAP-D
Big CHAP, deoxy
4.2
(1.4)
F2
OHES
Octyl-2-hydroxyethyl-sulfoxide
48.4
(24.2)
F3
RDHPOS
Rac-2,3-dihydroxypropyloctylsulfoxide
48.4
(24.2)
F4
GX-100
Genapol ® X-100
7.5
(0.15)
F5
HTG
n-Heptyl-β-D-thioglucopyranoside
58
(29)
F6
OG
n-Octyl-β-D-glucopyranoside
36
(18)
F7
NG
n-Nonyl-β-D-glucopyranoside
16.25
(6.5)
F8
CYGLU-3
CYGLU ®-3
56
(28)
F9
HECAMEG
HECAMEG
39
(19.5)
F10
HEGA-9
Hega ®-9
78
(39)
F11
C-HEGA-10
C-Hega ®-10
70
(35)
F12
C-HEGA-11
C-Hega ®-11
23
(11.5)
G1
CYMAL-3
CYMAL ®-3
60
(30)
G2
CYMAL-4
CYMAL ®-4
19
(7.6)
G3
CYMAL-5
CYMAL ®-5
7.2
(2.4)
G4
CYMAL-6
CYMAL ®-6
5.6
(0.56)
G5
CYMAL-7
CYMAL ®-7
9.5
(0.19)
G6
DMHM
2,6-Dimethyl-4-heptyl-β-D-maltoside
55
(27.5)
G7
OM
n-Octyl-β-D-maltopyranoside
39
(19.5)
G8
NM
n-Nonyl-β-D-maltopyranoside
15
(6)
G9
DαM
n-Decyl-α-D-maltopyranoside
4.8
(1.6)
G10
DM
n-Decyl-β-D-maltopyranoside
5.4
(1.8)
G11
UDαM
n-Undecyl-α-D-maltopyranoside
5.8
(0.58)
G12
UDM
n-Undecyl-β-D-maltopyranoside
5.9
(0.59)
H1
ωUDM
ω-Undecylenyl-β-D-maltopyranoside
3.6
(1.2)
H2
DDαM
n-Dodecyl-α-D-maltopyranoside
7.5
(0.15)
H3
DDM
n-Dodecyl-β-D-maltopyranoside
8.5
(0.17)
H4
TDM
n-Tridecyl-β-D-maltopyranoside
1.5
(0.03)
H5
OTM
n-Octyl-β-D-thiomaltopyranoside
21.25
(8.5)
H6
NTM
n-Nonyl-β-D-thiomaltopyranoside
9.6
(3.2)
H7
DTM
n-Decyl-β-D-thiomaltopyranoside
9
(0.9)
H8
UDTM
n-Undecyl-β-D-thiomaltopyranoside
10.5
(0.21)
H9
DDTM
n-Dodecyl-β-D-thiomaltopyranoside
5
(0.05)
H10
S-8
Sucrose8
48.8
(24.4)
H11
S-10
Sucrose10
7.5
(2.5)
H12
S-12
Sucrose12
15
(0.3)
16 . The method of claim 14 , wherein said detergents and controls are selected from the following table:
Well
Abbrev.
Name
[Det] mM
A1
No detergent (−control)
A2
Empty well for current detergent (+control)
A3
Z3-12
ZWITTERGENT ® 3-12
8.4
(2.8)
A4
Z3-14
ZWITTERGENT ® 3-14
10
(0.2)
A5
DMG
n-Decyl-N,N-dimethylglycine
38
(19)
A6
DOMG
n-Dodecyl-N,N-dimethylglycine
4.5
(1.5)
A7
DAO
n-Decyl-N,N-dimethylamine-N-oxide
21
(10.5)
A8
UDAO
n-Undecyl-N,N,-dimethylamine-N-oxide
9.6
(3.2)
A9
LDAO
n-Dodecyl-N,N-dimethylamine-N-oxide
3
(1)
A10
C-DDFOS
C-DODECAFOS ™
44
(22)
A11
CF-4
CYCLOFOS ™-4
28
(14)
A12
CF-5
CYCLOFOS ™-5
13.5
(4.5)
B1
CF-6
CYCLOFOS ™-6
8.04
(2.68)
B2
CF-7
CYCLOFOS ™-7
6.2
(0.62)
B3
FC-10
FOS-CHOLINE ®-10
22
(11)
B4
FC-11
FOS-CHOLINE ®-11
5.55
(1.85)
B5
FC-12
FOS-CHOLINE ®-12
4.5
(1.5)
B6
FC-13
FOS-CHOLINE ®-13
7.5
(0.75)
B7
FC-14
FOS-CHOLINE ®-14
6
(0.12)
B8
FC-I11
FOS-CHOLINE ®-ISO-11
53.2
(26.6)
B9
FC-I11-6U
FOS-CHOLINE ®-ISO-11-6U
51.6
(25.8)
B10
FC-I9
FOS-CHOLINE ®-ISO-9
64
(32)
B11
FC-U10-11
FOS-CHOLINE ®-UNSAT-11-10
15.5
(6.2)
B12
DHPC
1,2-Diheptanoyl-sn-glycero-3-phosphocholine
4.2
(1.4)
C1
LPC-10
LysoPC-10
20
(8)
C2
LPC-12
LysoPC-12
7
(0.7)
C3
FOSFEN-9
FOSFEN ™-9
4.05
(1.35)
C4
CHAPS
CHAPS
20
(8)
C5
CHAPSO
CHAPSO
20
(8)
C6
DDMAU
n-Dodecyl-N,N-(dimethylammonio)undecanoate
6.5
(0.13)
C7
DDMAB
n-Dodecyl-N,N-(dimethylammonio)butyrate
12.9
(4.3)
C8
LAPAO
LAPAO
4.8
(1.6)
C9
TRIPAO
TRIPAO
13.5
(4.5)
C10
T-20
TWEEN ® 20
5.9
(0.059)
C11
BRIJ-35
BRIJ ®35
9.1
(0.091)
C12
TX-100
TRITON ® X-100
11.5
(0.23)
D1
TX-114
TRITON ® X-114
10
(0.2)
D2
TX-305
TRITON ® X-305
6.5
(0.65)
D3
TX-405
TRITON ® X-405
8.1
(0.81)
D4
NID-P40
[Octylphenoxy]polyethoxyethanol
15
(0.3)
D5
APO8
Dimethyloctylphosphine oxide
80
(40)
D6
APO9
Dimethylnonylphosphine oxide
20
(10)
D7
APO10
Dimethyldecylphosphine oxide
13.98
(4.66)
D8
APO11
Dimethylundecylphosphine oxide
3.6
(1.2)
D9
APO12
Dimethyldodecylphosphine oxide
5.7
(0.57)
D10
C6E3
Triethylene glycol monohexyl ether
46
(23)
D11
C6E4
Tetraethylene glycol monohexyl ether
60
(30)
D12
C6E5
Pentaethylene glycol monohexyl ether
74
(37)
E1
C7E5
Pentaethylene glycol monoheptyl ether
42
(21)
E2
C8E4
Tetraethylene glycol monooctyl ether
20
(8)
E3
C8E5
Pentaethylene glycol monooctyl ether
17.75
(7.1)
E4
C8E6
Hexaethylene glycol monooctyl ether
25
(10)
E5
C10E5
Pentaethylene glycol monodecyl ether
8.1
(0.81)
E6
C10E6
Hexaethylene glycol monodecyl ether
9
(0.9)
E7
C10E9
Polyoxyethylene(9)decyl ether
3.9
(1.3)
E8
C12E8
Octaethylene glycol monododecyl ether
9
(0.09)
E9
C12E9
Polyoxyethylene(9)dodecyl ether
5
(0.05)
E10
C12E10
Polyoxyethylene(10)dodecyl ether
10
(0.2)
E11
C13E8
Polyoxyethylene(8)tridecyl ether
10
(0.1)
E12
CHAP
Big CHAP
8.7
(2.9)
F1
CHAP-D
Big CHAP, deoxy
4.2
(1.4)
F2
OHES
Octyl-2-hydroxyethyl-sulfoxide
48.4
(24.2)
F3
RDHPOS
Rac-2,3-dihydroxypropyloctylsulfoxide
48.4
(24.2)
F4
GX-100
Genapol ® X-100
7.5
(0.15)
F5
HTG
n-Heptyl-β-D-thioglucopyranoside
58
(29)
F6
OG
n-Octyl-β-D-glucopyranoside
36
(18)
F7
NG
n-Nonyl-β-D-glucopyranoside
16.25
(6.5)
F8
CYGLU-3
CYGLU ®-3
56
(28)
F9
HECAMEG
HECAMEG
39
(19.5)
F10
HEGA-9
Hega ®-9
78
(39)
F11
C-HEGA-10
C-Hega ®-10
70
(35)
F12
C-HEGA-11
C-Hega ®-11
23
(11.5)
G1
CYMAL-3
CYMAL ®-3
60
(30)
G2
CYMAL-4
CYMAL ®-4
19
(7.6)
G3
CYMAL-5
CYMAL ®-5
7.2
(2.4)
G4
CYMAL-6
CYMAL ®-6
5.6
(0.56)
G5
CYMAL-7
CYMAL ®-7
9.5
(0.19)
G6
DMHM
2,6-Dimethyl-4-heptyl-β-D-maltoside
55
(27.5)
G7
OM
n-Octyl-β-D-maltopyranoside
39
(19.5)
G8
NM
n-Nonyl-β-D-maltopyranoside
15
(6)
G9
DαM
n-Decyl-α-D-maltopyranoside
4.8
(1.6)
G10
DM
n-Decyl-β-D-maltopyranoside
5.4
(1.8)
G11
UDαM
n-Undecyl-α-D-maltopyranoside
5.8
(0.58)
G12
UDM
n-Undecyl-β-D-maltopyranoside
5.9
(0.59)
H1
ωUDM
ω-Undecylenyl-β-D-maltopyranoside
3.6
(1.2)
H2
DDαM
n-Dodecyl-α-D-maltopyranoside
7.5
(0.15)
H3
DDM
n-Dodecyl-β-D-maltopyranoside
8.5
(0.17)
H4
TDM
n-Tridecyl-β-D-maltopyranoside
1.5
(0.03)
H5
OTM
n-Octyl-β-D-thiomaltopyranoside
21.25
(8.5)
H6
NTM
n-Nonyl-β-D-thiomaltopyranoside
9.6
(3.2)
H7
DTM
n-Decyl-β-D-thiomaltopyranoside
9
(0.9)
H8
UDTM
n-Undecyl-β-D-thiomaltopyranoside
10.5
(0.21)
H9
DDTM
n-Dodecyl-β-D-thiomaltopyranoside
5
(0.05)
H10
S-8
Sucrose8
48.8
(24.4)
H11
S-10
Sucrose10
7.5
(2.5)
H12
S-12
Sucrose12
15
(0.3)
17 . The method of claim 1 , wherein said solution comprising said protein in a first detergent comprises about 1000 micrograms or less of said protein.
18 . The method of claim 1 , wherein said solution comprising said protein in a first detergent comprises about 500 micrograms or less of said protein.
19 . The method of claim 1 , wherein said solution comprising said protein in a first detergent comprises about 400 micrograms or less of said protein.
20 . The method of claim 1 , wherein said solution comprising said protein in a first detergent comprises about 200 micrograms or less of said protein.
21 . The method of claim 1 , wherein said solution comprising said protein in a first detergent comprises about 100 micrograms or less of said protein.
22 . The method of claim 1 , wherein said solution comprising said protein in a first detergent comprises about 50 micrograms or less of said protein.
23 . The method of claim 1 , wherein said method is performed in less than about two hours.
24 . The method of claim 1 , wherein said method is performed in less than about 1 hour.
25 . The method of claim 1 , wherein said protein amounts are determined by dot blot or Western blot analysis.
26 . The method of claim 25 , wherein said protein amounts determined from the high molecular weight cut-off dot blots are normalized and plotted with the ratio of low:high normalized intensities and the values grouped into quartiles.
27 . The method of claim 25 , wherein said protein amounts determined from the high molecular weight cut-off dot blots are normalized and plotted graphically on the abscissa while the ratio of low:high normalized intensities are plotted on the ordinate.
28 . The method of claim 1 , further wherein said method is used to screen detergent mixtures, additives, ionic strength, and pH.
29 . The method of claim 1 , wherein said protein is a membrane protein.
30 . The method of claim 1 , wherein the concentration of said different detergent is the critical micelle concentration of said different detergent.
31 . The method of claim 30 , wherein the critical micelle concentration of said different detergent is selected from the following table:
Well
Abbrev.
Name
[Det] mM
A1
No detergent (−control)
A2
Empty well for current detergent (+control)
A3
Z3-12
ZWITTERGENT ® 3-12
8.4
(2.8)
A4
Z3-14
ZWITTERGENT ® 3-14
10
(0.2)
A5
DMG
n-Decyl-N,N-dimethylglycine
38
(19)
A6
DOMG
n-Dodecyl-N,N-dimethylglycine
4.5
(1.5)
A7
DAO
n-Decyl-N,N-dimethylamine-N-oxide
21
(10.5)
A8
UDAO
n-Undecyl-N,N,-dimethylamine-N-oxide
9.6
(3.2)
A9
LDAO
n-Dodecyl-N,N-dimethylamine-N-oxide
3
(1)
A10
C-DDFOS
C-DODECAFOS ™
44
(22)
A11
CF-4
CYCLOFOS ™-4
28
(14)
A12
CF-5
CYCLOFOS ™-5
13.5
(4.5)
B1
CF-6
CYCLOFOS ™-6
8.04
(2.68)
B2
CF-7
CYCLOFOS ™-7
6.2
(0.62)
B3
FC-10
FOS-CHOLINE ®-10
22
(11)
B4
FC-11
FOS-CHOLINE ®-11
5.55
(1.85)
B5
FC-12
FOS-CHOLINE ®-12
4.5
(1.5)
B6
FC-13
FOS-CHOLINE ®-13
7.5
(0.75)
B7
FC-14
FOS-CHOLINE ®-14
6
(0.12)
B8
FC-I11
FOS-CHOLINE ®-ISO-11
53.2
(26.6)
B9
FC-I11-6U
FOS-CHOLINE ®-ISO-11-6U
51.6
(25.8)
B10
FC-I9
FOS-CHOLINE ®-ISO-9
64
(32)
B11
FC-U10-11
FOS-CHOLINE ®-UNSAT-11-10
15.5
(6.2)
B12
DHPC
1,2-Diheptanoyl-sn-glycero-3-phosphocholine
4.2
(1.4)
C1
LPC-10
LysoPC-10
20
(8)
C2
LPC-12
LysoPC-12
7
(0.7)
C3
FOSFEN-9
FOSFEN ™-9
4.05
(1.35)
C4
CHAPS
CHAPS
20
(8)
C5
CHAPSO
CHAPSO
20
(8)
C6
DDMAU
n-Dodecyl-N,N-(dimethylammonio)undecanoate
6.5
(0.13)
C7
DDMAB
n-Dodecyl-N,N-(dimethylammonio)butyrate
12.9
(4.3)
C8
LAPAO
LAPAO
4.8
(1.6)
C9
TRIPAO
TRIPAO
13.5
(4.5)
C10
T-20
TWEEN ® 20
5.9
(0.059)
C11
BRIJ-35
BRIJ ® 35
9.1
(0.091)
C12
TX-100
TRITON ® X-100
11.5
(0.23)
D1
TX-114
TRITON ® X-114
10
(0.2)
D2
TX-305
TRITON ® X-305
6.5
(0.65)
D3
TX-405
TRITON ® X-405
8.1
(0.81)
D4
NID-P40
[Octylphenoxy]polyethoxyethanol
15
(0.3)
D5
APO8
Dimethyloctylphosphine oxide
80
(40)
D6
APO9
Dimethylnonylphosphine oxide
20
(10)
D7
APO10
Dimethyldecylphosphine oxide
13.98
(4.66)
D8
APO11
Dimethylundecylphosphine oxide
3.6
(1.2)
D9
APO12
Dimethyldodecylphosphine oxide
5.7
(0.57)
D10
C6E3
Triethylene glycol monohexyl ether
46
(23)
D11
C6E4
Tetraethylene glycol monohexyl ether
60
(30)
D12
C6E5
Pentaethylene glycol monohexyl ether
74
(37)
E1
C7E5
Pentaethylene glycol monoheptyl ether
42
(21)
E2
C8E4
Tetraethylene glycol monooctyl ether
20
(8)
E3
C8E5
Pentaethylene glycol monooctyl ether
17.75
(7.1)
E4
C8E6
Hexaethylene glycol monooctyl ether
25
(10)
E5
C10E5
Pentaethylene glycol monodecyl ether
8.1
(0.81)
E6
C10E6
Hexaethylene glycol monodecyl ether
9
(0.9)
E7
C10E9
Polyoxyethylene(9)decyl ether
3.9
(1.3)
E8
C12E8
Octaethylene glycol monododecyl ether
9
(0.09)
E9
C12E9
Polyoxyethylene(9)dodecyl ether
5
(0.05)
E10
C12E10
Polyoxyethylene(10)dodecyl ether
10
(0.2)
E11
C13E8
Polyoxyethylene(8)tridecyl ether
10
(0.1)
E12
CHAP
Big CHAP
8.7
(2.9)
F1
CHAP-D
Big CHAP, deoxy
4.2
(1.4)
F2
OHES
Octyl-2-hydroxyethyl-sulfoxide
48.4
(24.2)
F3
RDHPOS
Rac-2,3-dihydroxypropyloctylsulfoxide
48.4
(24.2)
F4
GX-100
Genapol ® X-100
7.5
(0.15)
F5
HTG
n-Heptyl-β-D-thioglucopyranoside
58
(29)
F6
OG
n-Octyl-β-D-glucopyranoside
36
(18)
F7
NG
n-Nonyl-β-D-glucopyranoside
16.25
(6.5)
F8
CYGLU-3
CYGLU ®-3
56
(28)
F9
HECAMEG
HECAMEG
39
(19.5)
F10
HEGA-9
Hega ®-9
78
(39)
F11
C-HEGA-10
C-Hega ®-10
70
(35)
F12
C-HEGA-11
C-Hega ®-11
23
(11.5)
G1
CYMAL-3
CYMAL ®-3
60
(30)
G2
CYMAL-4
CYMAL ®-4
19
(7.6)
G3
CYMAL-5
CYMAL ®-5
7.2
(2.4)
G4
CYMAL-6
CYMAL ®-6
5.6
(0.56)
G5
CYMAL-7
CYMAL ®-7
9.5
(0.19)
G6
DMHM
2,6-Dimethyl-4-heptyl-β-D-maltoside
55
(27.5)
G7
OM
n-Octyl-β-D-maltopyranoside
39
(19.5)
G8
NM
n-Nonyl-β-D-maltopyranoside
15
(6)
G9
DαM
n-Decyl-α-D-maltopyranoside
4.8
(1.6)
G10
DM
n-Decyl-β-D-maltopyranoside
5.4
(1.8)
G11
UDαM
n-Undecyl-α-D-maltopyranoside
5.8
(0.58)
G12
UDM
n-Undecyl-β-D-maltopyranoside
5.9
(0.59)
H1
ωUDM
ω-Undecylenyl-β-D-maltopyranoside
3.6
(1.2)
H2
DDαM
n-Dodecyl-α-D-maltopyranoside
7.5
(0.15)
H3
DDM
n-Dodecyl-β-D-maltopyranoside
8.5
(0.17)
H4
TDM
n-Tridecyl-β-D-maltopyranoside
1.5
(0.03)
H5
OTM
n-Octyl-β-D-thiomaltopyranoside
21.25
(8.5)
H6
NTM
n-Nonyl-β-D-thiomaltopyranoside
9.6
(3.2)
H7
DTM
n-Decyl-β-D-thiomaltopyranoside
9
(0.9)
H8
UDTM
n-Undecyl-β-D-thiomaltopyranoside
10.5
(0.21)
H9
DDTM
n-Dodecyl-β-D-thiomaltopyranoside
5
(0.05)
H10
S-8
Sucrose8
48.8
(24.4)
H11
S-10
Sucrose10
7.5
(2.5)
H12
S-12
Sucrose12
15
(0.3)
32 . The method of claim 1 , wherein said different detergent has moderate or high aqueous solubility.
33 . The method of claim 1 , wherein said different detergent has zwitterionic or nonionic headgroups.
34 . A detergent panel for determining the stability and size of a protein, said panel comprising at least two detergents at or above their critical micelle concentrations, and optionally a positive control and a negative control, wherein said detergents and their critical micelle concentrations, or suitably higher concentrations, are selected from the following table:
Well
Abbrev.
Name
[Det] mM
A1
No detergent (−control)
A2
Empty well for current detergent (+control)
A3
Z3-12
ZWITTERGENT ® 3-12
8.4
(2.8)
A4
Z3-14
ZWITTERGENT ® 3-14
10
(0.2)
A5
DMG
n-Decyl-N,N-dimethylglycine
38
(19)
A6
DOMG
n-Dodecyl-N,N-dimethylglycine
4.5
(1.5)
A7
DAO
n-Decyl-N,N-dimethylamine-N-oxide
21
(10.5)
A8
UDAO
n-Undecyl-N,N,-dimethylamine-N-oxide
9.6
(3.2)
A9
LDAO
n-Dodecyl-N,N-dimethylamine-N-oxide
3
(1)
A10
C-DDFOS
C-DODECAFOS ™
44
(22)
A11
CF-4
CYCLOFOS ™-4
28
(14)
A12
CF-5
CYCLOFOS ™-5
13.5
(4.5)
B1
CF-6
CYCLOFOS ™-6
8.04
(2.68)
B2
CF-7
CYCLOFOS ™-7
6.2
(0.62)
B3
FC-10
FOS-CHOLINE ®-10
22
(11)
B4
FC-11
FOS-CHOLINE ®-11
5.55
(1.85)
B5
FC-12
FOS-CHOLINE ®-12
4.5
(1.5)
B6
FC-13
FOS-CHOLINE ®-13
7.5
(0.75)
B7
FC-14
FOS-CHOLINE ®-14
6
(0.12)
B8
FC-I11
FOS-CHOLINE ®-ISO-11
53.2
(26.6)
B9
FC-I11-6U
FOS-CHOLINE ®-ISO-11-6U
51.6
(25.8)
B10
FC-I9
FOS-CHOLINE ®-ISO-9
64
(32)
B11
FC-U10-11
FOS-CHOLINE ®-UNSAT-11-10
15.5
(6.2)
B12
DHPC
1,2-Diheptanoyl-sn-glycero-3-phosphocholine
4.2
(1.4)
C1
LPC-10
LysoPC-10
20
(8)
C2
LPC-12
LysoPC-12
7
(0.7)
C3
FOSFEN-9
FOSFEN ™-9
4.05
(1.35)
C4
CHAPS
CHAPS
20
(8)
C5
CHAPSO
CHAPSO
20
(8)
C6
DDMAU
n-Dodecyl-N,N-(dimethylammonio)undecanoate
6.5
(0.13)
C7
DDMAB
n-Dodecyl-N,N-(dimethylammonio)butyrate
12.9
(4.3)
C8
LAPAO
LAPAO
4.8
(1.6)
C9
TRIPAO
TRIPAO
13.5
(4.5)
C10
T-20
TWEEN ® 20
5.9
(0.059)
C11
BRIJ-35
BRIJ ®35
9.1
(0.091)
C12
TX-100
TRITON ® X-100
11.5
(0.23)
D1
TX-114
TRITON ® X-114
10
(0.2)
D2
TX-305
TRITON ® X-305
6.5
(0.65)
D3
TX-405
TRITON ® X-405
8.1
(0.81)
D4
NID-P40
[Octylphenoxy]polyethoxyethanol
15
(0.3)
D5
APO8
Dimethyloctylphosphine oxide
80
(40)
D6
APO9
Dimethylnonylphosphine oxide
20
(10)
D7
APO10
Dimethyldecylphosphine oxide
13.98
(4.66)
D8
APO11
Dimethylundecylphosphine oxide
3.6
(1.2)
D9
APO12
Dimethyldodecylphosphine oxide
5.7
(0.57)
D10
C6E3
Triethylene glycol monohexyl ether
46
(23)
D11
C6E4
Tetraethylene glycol monohexyl ether
60
(30)
D12
C6E5
Pentaethylene glycol monohexyl ether
74
(37)
E1
C7E5
Pentaethylene glycol monoheptyl ether
42
(21)
E2
C8E4
Tetraethylene glycol monooctyl ether
20
(8)
E3
C8E5
Pentaethylene glycol monooctyl ether
17.75
(7.1)
E4
C8E6
Hexaethylene glycol monooctyl ether
25
(10)
E5
C10E5
Pentaethylene glycol monodecyl ether
8.1
(0.81)
E6
C10E6
Hexaethylene glycol monodecyl ether
9
(0.9)
E7
C10E9
Polyoxyethylene(9)decyl ether
3.9
(1.3)
E8
C12E8
Octaethylene glycol monododecyl ether
9
(0.09)
E9
C12E9
Polyoxyethylene(9)dodecyl ether
5
(0.05)
E10
C12E10
Polyoxyethylene(10)dodecyl ether
10
(0.2)
E11
C13E8
Polyoxyethylene(8)tridecyl ether
10
(0.1)
E12
CHAP
Big CHAP
8.7
(2.9)
F1
CHAP-D
Big CHAP, deoxy
4.2
(1.4)
F2
OHES
Octyl-2-hydroxyethyl-sulfoxide
48.4
(24.2)
F3
RDHPOS
Rac-2,3-dihydroxypropyloctylsulfoxide
48.4
(24.2)
F4
GX-100
Genapol ® X-100
7.5
(0.15)
F5
HTG
n-Heptyl-β-D-thioglucopyranoside
58
(29)
F6
OG
n-Octyl-β-D-glucopyranoside
36
(18)
F7
NG
n-Nonyl-β-D-glucopyranoside
16.25
(6.5)
F8
CYGLU-3
CYGLU ®-3
56
(28)
F9
HECAMEG
HECAMEG
39
(19.5)
F10
HEGA-9
Hega ®-9
78
(39)
F11
C-HEGA-10
C-Hega ®-10
70
(35)
F12
C-HEGA-11
C-Hega ®-11
23
(11.5)
G1
CYMAL-3
CYMAL ®-3
60
(30)
G2
CYMAL-4
CYMAL ®-4
19
(7.6)
G3
CYMAL-5
CYMAL ®-5
7.2
(2.4)
G4
CYMAL-6
CYMAL ®-6
5.6
(0.56)
G5
CYMAL-7
CYMAL ®-7
9.5
(0.19)
G6
DMHM
2,6-Dimethyl-4-heptyl-β-D-maltoside
55
(27.5)
G7
OM
n-Octyl-β-D-maltopyranoside
39
(19.5)
G8
NM
n-Nonyl-β-D-maltopyranoside
15
(6)
G9
DαM
n-Decyl-α-D-maltopyranoside
4.8
(1.6)
G10
DM
n-Decyl-β-D-maltopyranoside
5.4
(1.8)
G11
UDαM
n-Undecyl-α-D-maltopyranoside
5.8
(0.58)
G12
UDM
n-Undecyl-β-D-maltopyranoside
5.9
(0.59)
H1
ωUDM
ω-Undecylenyl-β-D-maltopyranoside
3.6
(1.2)
H2
DDαM
n-Dodecyl-α-D-maltopyranoside
7.5
(0.15)
H3
DDM
n-Dodecyl-β-D-maltopyranoside
8.5
(0.17)
H4
TDM
n-Tridecyl-β-D-maltopyranoside
1.5
(0.03)
H5
OTM
n-Octyl-β-D-thiomaltopyranoside
21.25
(8.5)
H6
NTM
n-Nonyl-β-D-thiomaltopyranoside
9.6
(3.2)
H7
DTM
n-Decyl-β-D-thiomaltopyranoside
9
(0.9)
H8
UDTM
n-Undecyl-β-D-thiomaltopyranoside
10.5
(0.21)
H9
DDTM
n-Dodecyl-β-D-thiomaltopyranoside
5
(0.05)
H10
S-8
Sucrose8
48.8
(24.4)
H11
S-10
Sucrose10
7.5
(2.5)
H12
S-12
Sucrose12
15
(0.3)
35 . The detergent panel of claim 34 , wherein all 96 detergents and controls are used.
36 . A kit for determining the stability and size of a protein, said kit comprising at least one detergent, at least one chamber wherein said chamber comprises a filter of about 0.2 μm pore size, at least one chamber comprising a high molecular weight cut-off filter, at least one chamber comprising a low molecular weight cut-off filter, optionally at least one standard protein, and an instructional material for the used thereof.
37 . The kit of claim 36 , wherein said chambers are wells in 96 well plates, said high molecular weight cut-off filter is about 300 kDa, and said low molecular weight cut-off filter is about 100 kDa.
38 . The kit of claim 36 , further comprising at least one detergent or control selected from the following table:
Well
Abbrev.
Name
[Det] mM
A1
No detergent (−control)
A2
Empty well for current detergent (+control)
A3
Z3-12
ZWITTERGENT ® 3-12
8.4
(2.8)
A4
Z3-14
ZWITTERGENT ® 3-14
10
(0.2)
A5
DMG
n-Decyl-N,N-dimethylglycine
38
(19)
A6
DOMG
n-Dodecyl-N,N-dimethylglycine
4.5
(1.5)
A7
DAO
n-Decyl-N,N-dimethylamine-N-oxide
21
(10.5)
A8
UDAO
n-Undecyl-N,N,-dimethylamine-N-oxide
9.6
(3.2)
A9
LDAO
n-Dodecyl-N,N-dimethylamine-N-oxide
3
(1)
A10
C-DDFOS
C-DODECAFOS ™
44
(22)
A11
CF-4
CYCLOFOS ™-4
28
(14)
A12
CF-5
CYCLOFOS ™-5
13.5
(4.5)
B1
CF-6
CYCLOFOS ™-6
8.04
(2.68)
B2
CF-7
CYCLOFOS ™-7
6.2
(0.62)
B3
FC-10
FOS-CHOLINE ®-10
22
(11)
B4
FC-11
FOS-CHOLINE ®-11
5.55
(1.85)
B5
FC-12
FOS-CHOLINE ®-12
4.5
(1.5)
B6
FC-13
FOS-CHOLINE ®-13
7.5
(0.75)
B7
FC-14
FOS-CHOLINE ®-14
6
(0.12)
B8
FC-I11
FOS-CHOLINE ®-ISO-11
53.2
(26.6)
B9
FC-I11-6U
FOS-CHOLINE ®-ISO-11-6U
51.6
(25.8)
B10
FC-I9
FOS-CHOLINE ®-ISO-9
64
(32)
B11
FC-U10-11
FOS-CHOLINE ®-UNSAT-11-10
15.5
(6.2)
B12
DHPC
1,2-Diheptanoyl-sn-glycero-3-phosphocholine
4.2
(1.4)
C1
LPC-10
LysoPC-10
20
(8)
C2
LPC-12
LysoPC-12
7
(0.7)
C3
FOSFEN-9
FOSFEN ™-9
4.05
(1.35)
C4
CHAPS
CHAPS
20
(8)
C5
CHAPSO
CHAPSO
20
(8)
C6
DDMAU
n-Dodecyl-N,N-(dimethylammonio)undecanoate
6.5
(0.13)
C7
DDMAB
n-Dodecyl-N,N-(dimethylammonio)butyrate
12.9
(4.3)
C8
LAPAO
LAPAO
4.8
(1.6)
C9
TRIPAO
TRIPAO
13.5
(4.5)
C10
T-20
TWEEN ® 20
5.9
(0.059)
C11
BRIJ-35
BRIJ ®35
9.1
(0.091)
C12
TX-100
TRITON ® X-100
11.5
(0.23)
D1
TX-114
TRITON ® X-114
10
(0.2)
D2
TX-305
TRITON ® X-305
6.5
(0.65)
D3
TX-405
TRITON ® X-405
8.1
(0.81)
D4
NID-P40
[Octylphenoxy]polyethoxyethanol
15
(0.3)
D5
APO8
Dimethyloctylphosphine oxide
80
(40)
D6
APO9
Dimethylnonylphosphine oxide
20
(10)
D7
APO10
Dimethyldecylphosphine oxide
13.98
(4.66)
D8
APO11
Dimethylundecylphosphine oxide
3.6
(1.2)
D9
APO12
Dimethyldodecylphosphine oxide
5.7
(0.57)
D10
C6E3
Triethylene glycol monohexyl ether
46
(23)
D11
C6E4
Tetraethylene glycol monohexyl ether
60
(30)
D12
C6E5
Pentaethylene glycol monohexyl ether
74
(37)
E1
C7E5
Pentaethylene glycol monoheptyl ether
42
(21)
E2
C8E4
Tetraethylene glycol monooctyl ether
20
(8)
E3
C8E5
Pentaethylene glycol monooctyl ether
17.75
(7.1)
E4
C8E6
Hexaethylene glycol monooctyl ether
25
(10)
E5
C10E5
Pentaethylene glycol monodecyl ether
8.1
(0.81)
E6
C10E6
Hexaethylene glycol monodecyl ether
9
(0.9)
E7
C10E9
Polyoxyethylene(9)decyl ether
3.9
(1.3)
E8
C12E8
Octaethylene glycol monododecyl ether
9
(0.09)
E9
C12E9
Polyoxyethylene(9)dodecyl ether
5
(0.05)
E10
C12E10
Polyoxyethylene(10)dodecyl ether
10
(0.2)
E11
C13E8
Polyoxyethylene(8)tridecyl ether
10
(0.1)
E12
CHAP
Big CHAP
8.7
(2.9)
F1
CHAP-D
Big CHAP, deoxy
4.2
(1.4)
F2
OHES
Octyl-2-hydroxyethyl-sulfoxide
48.4
(24.2)
F3
RDHPOS
Rac-2,3-dihydroxypropyloctylsulfoxide
48.4
(24.2)
F4
GX-100
Genapol ® X-100
7.5
(0.15)
F5
HTG
n-Heptyl-β-D-thioglucopyranoside
58
(29)
F6
OG
n-Octyl-β-D-glucopyranoside
36
(18)
F7
NG
n-Nonyl-β-D-glucopyranoside
16.25
(6.5)
F8
CYGLU-3
CYGLU ®-3
56
(28)
F9
HECAMEG
HECAMEG
39
(19.5)
F10
HEGA-9
Hega ®-9
78
(39)
F11
C-HEGA-10
C-Hega ®-10
70
(35)
F12
C-HEGA-11
C-Hega ®-11
23
(11.5)
G1
CYMAL-3
CYMAL ®-3
60
(30)
G2
CYMAL-4
CYMAL ®-4
19
(7.6)
G3
CYMAL-5
CYMAL ®-5
7.2
(2.4)
G4
CYMAL-6
CYMAL ®-6
5.6
(0.56)
G5
CYMAL-7
CYMAL ®-7
9.5
(0.19)
G6
DMHM
2,6-Dimethyl-4-heptyl-β-D-maltoside
55
(27.5)
G7
OM
n-Octyl-β-D-maltopyranoside
39
(19.5)
G8
NM
n-Nonyl-β-D-maltopyranoside
15
(6)
G9
DαM
n-Decyl-α-D-maltopyranoside
4.8
(1.6)
G10
DM
n-Decyl-β-D-maltopyranoside
5.4
(1.8)
G11
UDαM
n-Undecyl-α-D-maltopyranoside
5.8
(0.58)
G12
UDM
n-Undecyl-β-D-maltopyranoside
5.9
(0.59)
H1
ωUDM
ω-Undecylenyl-β-D-maltopyranoside
3.6
(1.2)
H2
DDαM
n-Dodecyl-α-D-maltopyranoside
7.5
(0.15)
H3
DDM
n-Dodecyl-β-D-maltopyranoside
8.5
(0.17)
H4
TDM
n-Tridecyl-β-D-maltopyranoside
1.5
(0.03)
H5
OTM
n-Octyl-β-D-thiomaltopyranoside
21.25
(8.5)
H6
NTM
n-Nonyl-β-D-thiomaltopyranoside
9.6
(3.2)
H7
DTM
n-Decyl-β-D-thiomaltopyranoside
9
(0.9)
H8
UDTM
n-Undecyl-β-D-thiomaltopyranoside
10.5
(0.21)
H9
DDTM
n-Dodecyl-β-D-thiomaltopyranoside
5
(0.05)
H10
S-8
Sucrose8
48.8
(24.4)
H11
S-10
Sucrose10
7.5
(2.5)
H12
S-12
Sucrose12
15
(0.3)
39 . The kit of claim 38 , wherein said kit comprises at least one test 96 well plate comprising all of the detergents and controls according to said table.
40 . The method of claim 1 , wherein at least one of said first, second, or third chambers is subjected to centrifugation to enhance the filtration process.Join the waitlist — get patent alerts
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