US 2015/0087554 A1Published application

Polypeptide markers for the diagnosis of alzheimer's disease

AssigneeMosaiques Diagnostics and Therapeutics AG

InventorHarald Mischak

Life of this patent

Estimating the term…

Abstract

A method for the diagnosis of the probability of Alzheimer's disease in a subject patient comprising the step of determining the presence or absence of at least three polypeptide markers in a sample, wherein the at least three polypeptide markers are at least three different markers selected from markers 1 to 50 (frequency markers) and markers 51 to 279 (amplitude markers), wherein the diagnosis is based on a comparison and ranking of the frequency and/or amplitude of the at least three polypeptide markers to the same at least three polypeptide markers obtained from control subjects with and without Alzheimer's disease.

Claims6

Dependent claims

A method for diagnosing the probability of Alzheimer's disease in a subject patient, comprising:

obtaining a sample of bodily fluid from said patient, said sample selected from the group consisting of blood, urine, cerebrospinal fluid or any combinations of the foregoing;
separating said sample patient into a plurality of polypeptides;
identifying said plurality of polypeptides;
comparing said plurality of polypeptides to known polypeptide markers taken from control subjects with Alzheimer's disease and without Alzheimer's disease to obtain a subset of polypeptide markers from said plurality of polypeptides that substantially match said known polypeptide markers;
wherein said known polypeptide markers are characterized by the following molecular masses and migration times (CE time):
Mass
Migration
No.
[g/mol]
time [min]
1
1073.35
34.96
2
1073.35
35.9
3
1110.31
37.67
4
1111.3
34.97
5
1196.36
36.37
6
1265.63
27.22
7
1322.44
36.2
8
1387.07
20.42
9
1390.49
36.26
10
1423.57
29.85
11
1453.55
30.47
12
1490.43
35.48
13
1494.62
30.23
14
1819.86
23.51
15
1927.02
21
16
2128.97
26.03
17
2214.22
26.48
18
2313.26
30.32
19
2423.19
27.03
20
2428.11
27.16
21
2521.33
27.86
22
2644.31
21.1
23
2706.37
28.3
24
3068.46
30.03
25
3079.61
37.02
26
3153.38
35.65
27
3229.79
25.02
28
3438.77
23.49
29
3481.8
30.62
30
3567.61
23.88
31
3685.85
25.47
32
3921.84
29.49
33
4036.3
24.92
34
4038.22
20.53
35
4093.96
21.04
36
4145.22
24.51
37
4372.3
19.37
38
4377.95
30.81
39
4496.98
19.35
40
4516
19.37
41
4540.4
22
42
4586.98
19.38
43
4596.23
19.6
44
4810.95
19.77
45
5227.91
25.08
46
5582.16
24.3
47
6402.73
22.3
48
7749.32
19.97
49
7760.02
28.72
50
10671.25
28.92
51
824.48
19.92
52
840.49
19.92
53
846.38
25.96
54
897.41
25.86
55
911.3
34.1
56
950.57
24.13
57
973.26
35.47
58
984.45
26.32
59
988.57
35.65
60
1013.43
25.18
61
1016.3
35.53
62
1041.58
36.79
63
1053.52
25.25
64
1055.61
25.7
65
1082.55
23.75
66
1082.65
26.15
67
1083.5
27.14
68
1083.55
25.61
69
1085.63
36.39
70
1094.61
25.99
71
1130.37
35.19
72
1130.63
27.07
73
1135.56
27.03
74
1150.61
26.55
75
1158.62
27.02
76
1183.66
27.49
77
1186.56
27.21
78
1217.56
21.8
79
1229.62
27.38
80
1250.61
27.59
81
1276.43
35.7
82
1283.4
35.91
83
1300.67
29.7
84
1306.74
22.16
85
1333.44
35.93
86
1342.44
35.43
87
1347.71
28.94
88
1349.67
28.25
89
1360.67
28.06
90
1386.68
28.62
91
1388.74
27.97
92
1399.44
36.06
93
1404.73
29.49
94
1409.63
22.11
95
1449.7
22.04
96
1451.72
28.84
97
1464.7
30.57
99
1475.77
30.05
100
1499.74
29.89
101
1519.76
31.87
102
1535.74
30
103
1542.78
23.24
104
1550.67
27.47
105
1552.61
30.78
106
1590.81
32.46
107
1596.83
30.32
108
1614.84
31.54
109
1623.65
31.1
110
1628.8
20.75
111
1636.79
22.71
112
1659.81
27.41
113
1692.91
31.38
114
1703.89
33.01
115
1727.92
32.13
116
1732.88
31.36
117
1738.68
32.16
118
1741.83
30.56
119
1756.89
19.81
120
1771.97
30.02
121
1800.02
25
122
1816.98
33.53
123
1820.06
32.23
124
1841.77
36.26
125
1849.68
18.31
126
1850.96
31.97
127
1867.72
33.07
128
1869.92
31.28
129
1873
21.26
130
1900.07
24.19
131
1914.07
25.67
132
1915.72
34.24
133
1917.03
24.99
134
1955.95
27.81
135
2021.11
21.85
136
2041.99
32.56
137
2053.18
20.54
138
2072.15
27.04
139
2077.07
25.44
140
2085.12
33.75
141
2140.12
20.56
142
2166.09
27.84
143
2168.66
34.88
144
2179.03
25.31
145
2278.11
26.17
146
2283.18
26.31
147
2314.3
22.71
148
2320.43
23.21
149
2327.24
34.9
150
2341.12
26.74
151
2343.24
34.95
152
2368.24
21.24
153
2377.25
28.07
154
2390.26
27.33
155
2392.73
35.48
156
2409.28
27.64
157
2410.3
30.62
158
2448.39
21.07
159
2475.21
27.09
160
2475.27
24.74
161
2486.04
35.76
162
2500.33
19.99
163
2521.36
19.26
164
2522.35
27.9
165
2527.42
19.79
166
2549.29
27.66
167
2584.37
35.6
168
2684.32
21.38
169
2698.41
22.52
170
2799.17
25.07
171
2816.31
28.74
172
2851.42
27.7
173
2923.51
36.73
174
2982.72
19.97
175
3019.54
24.72
176
3173.68
22.95
177
3192.67
37.66
178
3215.61
28.53
179
3218.4
30.31
180
3221.59
22.7
181
3279.66
25.32
182
3282.34
36.05
183
3298.42
36.17
184
3302.68
23.31
185
3324.7
21.62
186
3386.77
22.21
187
3401.73
23.48
188
3408.76
26.04
189
3462.68
21.25
190
3466.75
23.9
191
3524.8
31.72
192
3529.84
23.7
193
3595.77
23.89
194
3605.4
21.53
195
3614.82
22.01
196
3644.79
29.39
197
3645.25
26.69
198
3650.74
19.9
199
3672.71
21.9
200
3677.74
24.3
201
3685.86
22.21
202
3687.78
20.58
203
3693.93
27.15
204
3741.63
20.1
205
3768.77
31.84
206
3782.94
22.68
207
3788.04
25.24
208
3792.76
27.25
209
3819.82
32.74
210
3881.87
24.56
211
3903.08
25.71
212
3905.8
27.72
213
3929.73
23.07
214
3934.76
24.02
215
3951.07
22.77
216
4005.54
22.57
217
4087.12
26.92
218
4112.78
24.46
219
4131.68
22.54
220
4164.86
25.11
221
4192.99
34.95
222
4203.13
26.14
223
4218.31
20.6
224
4232.32
32.35
225
4250
27.68
226
4278
25.3
227
4301.93
21.26
228
4307.09
31.58
229
4349.06
28.17
230
4384.07
19.8
231
4428.79
32.86
232
4464.64
21.54
233
4468.61
23.47
234
4510.97
28.56
235
4548.03
25.95
236
4562
29.31
237
4562
29.31
238
4564.18
26.01
239
4583.84
24.2
240
4637.81
27.82
241
4730.71
19.63
242
4749.41
21.81
243
4750.06
19.53
244
4890.59
23.27
245
4933.64
20.57
246
4986.98
21.35
247
5039.63
25.68
248
5042.1
24.31
249
5060.4
20.09
250
5110.36
25.28
251
5122.44
20.75
252
5157.79
34.26
253
5181.63
25.36
254
5258.56
22.09
255
5439.46
25.41
256
5495.33
31.72
257
5660.6
31.55
258
5745.2
19.72
259
5891.27
24.16
260
6224.17
25.27
261
6327.46
22.03
262
6498.75
20.1
263
6611.97
28.44
264
6617.76
24.03
265
6813.98
23.14
266
6881.81
21.72
267
6893.8
24.57
268
6985.31
21.87
269
7026.99
22.02
270
7049.04
22.79
271
7106.83
23
272
7142.49
21.32
273
7210.86
22.94
274
8176.16
19.47
275
8186.93
20.72
276
8289.28
19.39
277
9724.01
25.64
278
12529
20.64
279
14557.83
19.21
wherein said CE times are based on capillary electrophoresis using a glass capillary of 90 cm in length with an inner diameter (ID) of 50 μm at an applied voltage of 30 kV, wherein 30% methanol and 0.5% formic acid in water is used as the mobile solvent for the capillary electrophoresis at a flow rate of 2 μl/min., and
wherein said CE times are calibrated relative to the following values:
Protein/Polypeptide
Migration time (minutes)
Aprotinin
9.2
Ribonuclease
10.9
Lysozyme
8.9
“REV” SEQ ID NO:
1
15.6
“ELM” SEQ ID NO:
2
23.4
“KINCON” SEQ ID NO:
3
20.0
“GIVLY” SEQ ID NO:
4
36.8;
determining if said subset of polypeptide markers comprises at least a first, second and third biomarker selected from:
a first biomarker that is at least one selected from the group consisting of:
(1) markers 1 to 50 (frequency markers) having (a) a frequency of presence in patients with Alzheimer's disease of at least 0.6 and (b) a frequency of presence in patients with Alzheimer's disease at least two (2) times the frequency of presence in patients without Alzheimer's disease;
(2) markers 1 to 50 (frequency markers) having (a) a frequency of presence in patients without Alzheimer's disease of at least 0.6 and (b) a frequency of presence in patients without Alzheimer's disease at least two (2) times the frequency of presence in patients without Alzheimer's disease;
(3) markers 51 to 279 (amplitude markers) having a mean amplitude in patients with Alzheimer's disease of at least 300 and an amplitude greater than in patients without Alzheimer's disease;
and (4) markers 51 to 279 (amplitude markers) having a mean amplitude in patients without Alzheimer's disease of at least 300 and an amplitude greater than in patients with Alzheimer's disease,
a second biomarker which is different than said first biomarker and is at least one selected from the group consisting of:
(1) markers 1 to 50 (frequency markers) having (a) a frequency of presence in patients with Alzheimer's disease of at least 0.6 and (b) a frequency of presence in patients with Alzheimer's disease at least two (2) times the frequency of presence in patients without Alzheimer's disease;
(2) markers 1 to 50 (frequency markers) having (a) a frequency of presence in patients without Alzheimer's disease of at least 0.6 and (b) a frequency of presence in patients without Alzheimer's disease at least two (2) times the frequency of presence in patients without Alzheimer's disease;
(3) markers 51 to 279 (amplitude markers) having a mean amplitude in patients with Alzheimer's disease of at least 300 and an amplitude greater than in patients without Alzheimer's disease;
and (4) markers 51 to 279 (amplitude markers) having a mean amplitude in patients without Alzheimer's disease of at least 300 and an amplitude greater than in patients with Alzheimer's disease, and
a third biomarker which is different than said first and second biomarkers and is at least one selected from the group consisting of markers 1 to 242 (frequency markers);
comparing the frequency and/or amplitude of presence of said at least said first, second and third biomarkers in said sample of said subject patient to the frequency and/or amplitude of presence of the same first, second and third biomarkers from said known polypeptide markers from said control subjects,
ranking said subject patient between said control subjects with Alzheimer's disease and without Alzheimer's disease based on the second comparing step;
and
diagnosing the probability of Alzheimer's disease in said subject patient based on said ranking.

879 more elements

Dependencies are inferred from the claim text.

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Abstract

A method for diagnosing the probability of Alzheimer's disease in a subject patient, comprising:

obtaining a sample of bodily fluid from said patient, said sample selected from the group consisting of blood, urine, cerebrospinal fluid or any combinations of the foregoing;
separating said sample patient into a plurality of polypeptides;
identifying said plurality of polypeptides;
comparing said plurality of polypeptides to known polypeptide markers taken from control subjects with Alzheimer's disease and without Alzheimer's disease to obtain a subset of polypeptide markers from said plurality of polypeptides that substantially match said known polypeptide markers;
wherein said known polypeptide markers are characterized by the following molecular masses and migration times (CE time):
Mass
Migration
No.
[g/mol]
time [min]
1
1073.35
34.96
2
1073.35
35.9
3
1110.31
37.67
4
1111.3
34.97
5
1196.36
36.37
6
1265.63
27.22
7
1322.44
36.2
8
1387.07
20.42
9
1390.49
36.26
10
1423.57
29.85
11
1453.55
30.47
12
1490.43
35.48
13
1494.62
30.23
14
1819.86
23.51
15
1927.02
21
16
2128.97
26.03
17
2214.22
26.48
18
2313.26
30.32
19
2423.19
27.03
20
2428.11
27.16
21
2521.33
27.86
22
2644.31
21.1
23
2706.37
28.3
24
3068.46
30.03
25
3079.61
37.02
26
3153.38
35.65
27
3229.79
25.02
28
3438.77
23.49
29
3481.8
30.62
30
3567.61
23.88
31
3685.85
25.47
32
3921.84
29.49
33
4036.3
24.92
34
4038.22
20.53
35
4093.96
21.04
36
4145.22
24.51
37
4372.3
19.37
38
4377.95
30.81
39
4496.98
19.35
40
4516
19.37
41
4540.4
22
42
4586.98
19.38
43
4596.23
19.6
44
4810.95
19.77
45
5227.91
25.08
46
5582.16
24.3
47
6402.73
22.3
48
7749.32
19.97
49
7760.02
28.72
50
10671.25
28.92
51
824.48
19.92
52
840.49
19.92
53
846.38
25.96
54
897.41
25.86
55
911.3
34.1
56
950.57
24.13
57
973.26
35.47
58
984.45
26.32
59
988.57
35.65
60
1013.43
25.18
61
1016.3
35.53
62
1041.58
36.79
63
1053.52
25.25
64
1055.61
25.7
65
1082.55
23.75
66
1082.65
26.15
67
1083.5
27.14
68
1083.55
25.61
69
1085.63
36.39
70
1094.61
25.99
71
1130.37
35.19
72
1130.63
27.07
73
1135.56
27.03
74
1150.61
26.55
75
1158.62
27.02
76
1183.66
27.49
77
1186.56
27.21
78
1217.56
21.8
79
1229.62
27.38
80
1250.61
27.59
81
1276.43
35.7
82
1283.4
35.91
83
1300.67
29.7
84
1306.74
22.16
85
1333.44
35.93
86
1342.44
35.43
87
1347.71
28.94
88
1349.67
28.25
89
1360.67
28.06
90
1386.68
28.62
91
1388.74
27.97
92
1399.44
36.06
93
1404.73
29.49
94
1409.63
22.11
95
1449.7
22.04
96
1451.72
28.84
97
1464.7
30.57
99
1475.77
30.05
100
1499.74
29.89
101
1519.76
31.87
102
1535.74
30
103
1542.78
23.24
104
1550.67
27.47
105
1552.61
30.78
106
1590.81
32.46
107
1596.83
30.32
108
1614.84
31.54
109
1623.65
31.1
110
1628.8
20.75
111
1636.79
22.71
112
1659.81
27.41
113
1692.91
31.38
114
1703.89
33.01
115
1727.92
32.13
116
1732.88
31.36
117
1738.68
32.16
118
1741.83
30.56
119
1756.89
19.81
120
1771.97
30.02
121
1800.02
25
122
1816.98
33.53
123
1820.06
32.23
124
1841.77
36.26
125
1849.68
18.31
126
1850.96
31.97
127
1867.72
33.07
128
1869.92
31.28
129
1873
21.26
130
1900.07
24.19
131
1914.07
25.67
132
1915.72
34.24
133
1917.03
24.99
134
1955.95
27.81
135
2021.11
21.85
136
2041.99
32.56
137
2053.18
20.54
138
2072.15
27.04
139
2077.07
25.44
140
2085.12
33.75
141
2140.12
20.56
142
2166.09
27.84
143
2168.66
34.88
144
2179.03
25.31
145
2278.11
26.17
146
2283.18
26.31
147
2314.3
22.71
148
2320.43
23.21
149
2327.24
34.9
150
2341.12
26.74
151
2343.24
34.95
152
2368.24
21.24
153
2377.25
28.07
154
2390.26
27.33
155
2392.73
35.48
156
2409.28
27.64
157
2410.3
30.62
158
2448.39
21.07
159
2475.21
27.09
160
2475.27
24.74
161
2486.04
35.76
162
2500.33
19.99
163
2521.36
19.26
164
2522.35
27.9
165
2527.42
19.79
166
2549.29
27.66
167
2584.37
35.6
168
2684.32
21.38
169
2698.41
22.52
170
2799.17
25.07
171
2816.31
28.74
172
2851.42
27.7
173
2923.51
36.73
174
2982.72
19.97
175
3019.54
24.72
176
3173.68
22.95
177
3192.67
37.66
178
3215.61
28.53
179
3218.4
30.31
180
3221.59
22.7
181
3279.66
25.32
182
3282.34
36.05
183
3298.42
36.17
184
3302.68
23.31
185
3324.7
21.62
186
3386.77
22.21
187
3401.73
23.48
188
3408.76
26.04
189
3462.68
21.25
190
3466.75
23.9
191
3524.8
31.72
192
3529.84
23.7
193
3595.77
23.89
194
3605.4
21.53
195
3614.82
22.01
196
3644.79
29.39
197
3645.25
26.69
198
3650.74
19.9
199
3672.71
21.9
200
3677.74
24.3
201
3685.86
22.21
202
3687.78
20.58
203
3693.93
27.15
204
3741.63
20.1
205
3768.77
31.84
206
3782.94
22.68
207
3788.04
25.24
208
3792.76
27.25
209
3819.82
32.74
210
3881.87
24.56
211
3903.08
25.71
212
3905.8
27.72
213
3929.73
23.07
214
3934.76
24.02
215
3951.07
22.77
216
4005.54
22.57
217
4087.12
26.92
218
4112.78
24.46
219
4131.68
22.54
220
4164.86
25.11
221
4192.99
34.95
222
4203.13
26.14
223
4218.31
20.6
224
4232.32
32.35
225
4250
27.68
226
4278
25.3
227
4301.93
21.26
228
4307.09
31.58
229
4349.06
28.17
230
4384.07
19.8
231
4428.79
32.86
232
4464.64
21.54
233
4468.61
23.47
234
4510.97
28.56
235
4548.03
25.95
236
4562
29.31
237
4562
29.31
238
4564.18
26.01
239
4583.84
24.2
240
4637.81
27.82
241
4730.71
19.63
242
4749.41
21.81
243
4750.06
19.53
244
4890.59
23.27
245
4933.64
20.57
246
4986.98
21.35
247
5039.63
25.68
248
5042.1
24.31
249
5060.4
20.09
250
5110.36
25.28
251
5122.44
20.75
252
5157.79
34.26
253
5181.63
25.36
254
5258.56
22.09
255
5439.46
25.41
256
5495.33
31.72
257
5660.6
31.55
258
5745.2
19.72
259
5891.27
24.16
260
6224.17
25.27
261
6327.46
22.03
262
6498.75
20.1
263
6611.97
28.44
264
6617.76
24.03
265
6813.98
23.14
266
6881.81
21.72
267
6893.8
24.57
268
6985.31
21.87
269
7026.99
22.02
270
7049.04
22.79
271
7106.83
23
272
7142.49
21.32
273
7210.86
22.94
274
8176.16
19.47
275
8186.93
20.72
276
8289.28
19.39
277
9724.01
25.64
278
12529
20.64
279
14557.83
19.21
wherein said CE times are based on capillary electrophoresis using a glass capillary of 90 cm in length with an inner diameter (ID) of 50 μm at an applied voltage of 30 kV, wherein 30% methanol and 0.5% formic acid in water is used as the mobile solvent for the capillary electrophoresis at a flow rate of 2 μl/min., and
wherein said CE times are calibrated relative to the following values:
Protein/Polypeptide
Migration time (minutes)
Aprotinin
9.2
Ribonuclease
10.9
Lysozyme
8.9
“REV” SEQ ID NO:
1
15.6
“ELM” SEQ ID NO:
2
23.4
“KINCON” SEQ ID NO:
3
20.0
“GIVLY” SEQ ID NO:
4
36.8;
determining if said subset of polypeptide markers comprises at least a first, second and third biomarker selected from:
a first biomarker that is at least one selected from the group consisting of:
(1) markers 1 to 50 (frequency markers) having (a) a frequency of presence in patients with Alzheimer's disease of at least 0.6 and (b) a frequency of presence in patients with Alzheimer's disease at least two (2) times the frequency of presence in patients without Alzheimer's disease;
(2) markers 1 to 50 (frequency markers) having (a) a frequency of presence in patients without Alzheimer's disease of at least 0.6 and (b) a frequency of presence in patients without Alzheimer's disease at least two (2) times the frequency of presence in patients without Alzheimer's disease;
(3) markers 51 to 279 (amplitude markers) having a mean amplitude in patients with Alzheimer's disease of at least 300 and an amplitude greater than in patients without Alzheimer's disease;
and (4) markers 51 to 279 (amplitude markers) having a mean amplitude in patients without Alzheimer's disease of at least 300 and an amplitude greater than in patients with Alzheimer's disease,
a second biomarker which is different than said first biomarker and is at least one selected from the group consisting of:
(1) markers 1 to 50 (frequency markers) having (a) a frequency of presence in patients with Alzheimer's disease of at least 0.6 and (b) a frequency of presence in patients with Alzheimer's disease at least two (2) times the frequency of presence in patients without Alzheimer's disease;
(2) markers 1 to 50 (frequency markers) having (a) a frequency of presence in patients without Alzheimer's disease of at least 0.6 and (b) a frequency of presence in patients without Alzheimer's disease at least two (2) times the frequency of presence in patients without Alzheimer's disease;
(3) markers 51 to 279 (amplitude markers) having a mean amplitude in patients with Alzheimer's disease of at least 300 and an amplitude greater than in patients without Alzheimer's disease;
and (4) markers 51 to 279 (amplitude markers) having a mean amplitude in patients without Alzheimer's disease of at least 300 and an amplitude greater than in patients with Alzheimer's disease, and
a third biomarker which is different than said first and second biomarkers and is at least one selected from the group consisting of markers 1 to 242 (frequency markers);
comparing the frequency and/or amplitude of presence of said at least said first, second and third biomarkers in said sample of said subject patient to the frequency and/or amplitude of presence of the same first, second and third biomarkers from said known polypeptide markers from said control subjects,
ranking said subject patient between said control subjects with Alzheimer's disease and without Alzheimer's disease based on the second comparing step;
and
diagnosing the probability of Alzheimer's disease in said subject patient based on said ranking.

879 more elements