phase diagram
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Examples of Phase DiagramsExamples of Phase Diagrams
0 10 20 30 40 50 60 70 80 90
0.1
0.2
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0.4
0.5
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0.8
0.9
1.0
1.1
0 10 20 30 40 50 60 70 80 90
0.0
0.1
0.2
0.3
0.4
0.5
0.6
0.7
0.8
0.9
1.0
1.1
0 10 20 30 40 50 60 70 80 90
338
339
340
341
342
343
344
345
346
20 30 40 50 60 70
0.8
1.0
1.2
1.4
1.6
1.8
2.0
2.2
2.4
2.6
2.8
3.0
3.2
0 10 20 30 40 50 60 70 80 90
0.0
0.2
0.4
0.6
0.8
1.0
0 10 20 30 40 50 60 70 80 90
496
498
500
502
504
506
508
510
512
514
516
518
520
522
524
0 10 20 30 40 50 60 70 80 90
0.0
0.1
0.2
0.3
0.4
0.5
0.6
0 10 20 30 40 50 60 70 80 90
0.0
0.2
0.4
0.6
0.8
1.0
0 10 20 30 40 50 60 70 80 90 100
6
8
10
12
14
16
A
Re
lativ
e M
E a
t 2
22
nm
Temperature (oC)
B
Re
lativ
e F
l. p
ea
k in
ten
sity
Temperature (oC)
C
Fl.
pe
ak
po
sitio
n (
nm
)
Temperature (oC)
D
Re
lativ
e h
elix
co
nte
nt
Temperature (oC)
E
Re
lativ
e A
NS
Fl.
pe
ak
inte
nsi
ty
Temperature (oC)
F
AN
S F
l. p
ea
k p
osi
tion
(n
m)
Temperature (oC)
G
Op
tica
l de
nsi
ty a
t 3
50
nm
Temperature (oC)
H
Re
lativ
e F
l. lig
ht
sca
tte
rin
g
Temperature (oC)
Figure 1
I
Exc
ess
MH
C
Temperature (oC)
0 10 20 30 40 50 60 70 80 90
0.2
0.3
0.4
0.5
0.6
0.7
0.8
0.9
1.0
1.1
0 10 20 30 40 50 60 70 80 90
0.0
0.1
0.2
0.3
0.4
0.5
0.6
0.7
0.8
0.9
1.0
1.1
0 10 20 30 40 50 60 70 80 90
339
340
341
342
343
344
345
346
347
348
349
350
351
20 30 40 50 60 701.0
1.2
1.4
1.6
1.8
2.0
2.2
2.4
2.6
0 10 20 30 40 50 60 70 80 900.0
0.2
0.4
0.6
0.8
1.0
0 10 20 30 40 50 60 70 80 90
500
502
504
506
508
510
512
514
516
518
520
522
524
0 10 20 30 40 50 60 70 80 900.0
0.2
0.4
0.6
0.8
1.0
0 10 20 30 40 50 60 70 80 90
0.0
0.2
0.4
0.6
0.8
1.0
0 10 20 30 40 50 60 70 80 90 100
0
2
4
6
8
10
12
14
16
18
20
A
Re
lativ
e M
E a
t 2
22
nm
Temperature (oC)
B
Re
lativ
e F
l. p
ea
k in
ten
sity
Temperature (oC)
C
Fl.
pe
ak
po
sitio
n (
nm
)
Temperature (oC)
D
Re
lativ
e h
elix
co
nte
nt
Temperature (oC)
E
Re
lativ
e A
NS
Fl.
pe
ak
inte
nsi
ty
Temperature (oC)
F
AN
S F
l. p
ea
k p
osi
tion
(n
m)
Temperature (oC)
G
Op
tica
l de
nsi
ty a
t 3
50
nm
Temperature (oC)
H
Re
lativ
e F
l. lig
ht
sca
tte
rin
g
Temperature (oC)
Figure 2
I
Exc
ess
MH
C
Temperature (oC)
Interpretation
Experimental data Experimental data visualization visualization
VisualizationVisualization
Reconstructed 3-D Vector 321, xxxv i TpH
Rescale the vector to RGB color system
10 '''' '321, i
iTpH xxxxv
Each vector will define a color block in the phase diagram at its corresponding coordinates (pH, T)
'
'
'
3
2
1
x
x
x
B
G
R
Phase diagram created using high-resolution Phase diagram created using high-resolution second derivative UV spectroscopy data. second derivative UV spectroscopy data.
4 5 6 7 8pH
10
20
30
40
50
T Ricin2nd DerivativeUV
Phase diagram generated using ANS fluorescence and Phase diagram generated using ANS fluorescence and intrinsic Trp fluorescence intensity data and CD molar intrinsic Trp fluorescence intensity data and CD molar
ellipticity data of recombinant ricin toxin A-chain. ellipticity data of recombinant ricin toxin A-chain.
4 5 6 7 8 9pH
20
40
60
80
T Ricin ANS Int., Intrinsic Int., & CD
Phase diagram rPA based on intrinsic, ANS Phase diagram rPA based on intrinsic, ANS binding fluorescence and CDbinding fluorescence and CD
3 4 5 6 7 8 9pH
20
40
60
80
T Antrax PA # 1
2
Stable phase
Structrally altered and aggregated state
Molten globule state
Severely structurally altered phase
Empirical phase diagram (EPD) of Empirical phase diagram (EPD) of RSV over the pH range 3-8RSV over the pH range 3-8
Data included in the generation of the EPD are all negative second-derivative UV peaks, CD signal at 222 nm, optical density at 350 nm, intrinsic fluorescence peak position, intrinsic fluorescence intensity at 330 nm, and ANS fluorescence intensity at 485 nm
Ausar, et al, Mol. Pharm., 2 (6), 491 -499, 2005.
Adenovirus Type 2Adenovirus Type 2
Non-Viral Gene Delivery Vehicle Non-Viral Gene Delivery Vehicle DOTAPDOTAP
Non-Viral Gene Delivery Vehicle DOTAP Non-Viral Gene Delivery Vehicle DOTAP (Charge ratio =4.0)(Charge ratio =4.0)
Non-Viral Gene Delivery Vehicle Non-Viral Gene Delivery Vehicle DOTAP/DOPEDOTAP/DOPE
Non-Viral Gene Delivery Vehicle Non-Viral Gene Delivery Vehicle DOTAP/DOPE (Charge ratio = 4.0)DOTAP/DOPE (Charge ratio = 4.0)
Non-Viral Gene Delivery Vehicle PLL Non-Viral Gene Delivery Vehicle PLL (Charge ratio =4.0)(Charge ratio =4.0)
Ty21a bacteriaTy21a bacteria
HSPPC96HSPPC96
HSPPC96 Visual InterpretationHSPPC96 Visual Interpretation
HSC70HSC70
HSC70 Visual InterpretationHSC70 Visual Interpretation
FGF1FGF1
FGF1, heparin excipientFGF1, heparin excipient
FGF1, sucrose excipientFGF1, sucrose excipient
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