Discovery of N-glycan Biomarkers for the Canine Osteoarthritis
Abstract
:1. Introduction
2. Materials and Methods
2.1. Chemical
2.2. Dog Serum Samples
2.3. Serum N-glycan Release
2.4. Purification and Enrichment
2.5. Analysis of N-glycan by Nano-LC Chip Q-TOF MS and MS/MS
2.6. Data Processing and Statistical Analysis
3. Results and Discussion
3.1. MS Data Analysis for Serum N-glycan Annotation
3.2. Global Comparison of N-glycan Profile
3.3. Discovery of Potential N-glycan Biomarkers
3.4. Efficacy of N-glycan Biomarkers
4. Conclusions
Supplementary Materials
Author Contributions
Funding
Conflicts of Interest
References
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Healthy Control | OA Patient | Shared a | ||
---|---|---|---|---|
Number of Samples | 41 | 92 | ||
Number of N-glycan compositions | Extracted | 317 | 598 | |
Filtrated (f > 10%) b | 163 | 463 | 160 | |
Major class (Acc. Int. ≤ 95%) c | 36 (22.1%) | 50 (10.8%) | 35 | |
Minor class (Acc. Int. 95~99%) | 30 (18.4%) | 102 (22.0%) | 18 | |
Trace (Acc. Int. ≥ 9 9%) | 97 (59.5%) | 311 (67.2%) | 45 |
m/z | N-glycan a | Healthy Control | OA Patient | p-Value | ||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|
fb | AVE ± SD | CV | Rank | Acc. c | fb | AVE ± SD | CV | Rank | Acc. c | |||
910.3277 | 3_2_0_0_0 | 100% | 102.3 ± 14.9 | 14.6% | 22 | 84.1% | 100% | 33.8 ± 8.5 | 25.1% | 45 | 93.2% | 2.7 × 10−66 |
1056.3856 | 3_2_1_0_0 | 100% | 106.6 ± 13.6 | 12.7% | 17 | 78.9% | 100% | 34.7 ± 12.4 | 35.7% | 44 | 92.8% | 1.4 × 10−60 |
1754.6602 | 3_4_3_0_0 | 0% | 0.0 ± 0.0 | 354 | 100.0% | 100% | 24.3 ± 7.9 | 32.6% | 49 | 94.2% | 1.0 × 10−40 | |
1916.7130 | 4_4_3_0_0 | 39% | 1.2 ± 2.3 | 191.9% | 95 | 99.6% | 100% | 35.0 ± 11.7 | 33.5% | 43 | 92.5% | 8.0 × 10−38 |
2368.8408 | 5_4_1_2_0 | 100% | 507.0 ± 130.8 | 25.8% | 6 | 60.6% | 100% | 160.0 ± 86.1 | 53.8% | 14 | 70.4% | 1.5 × 10−37 |
2296.8197 | 6_5_0_1_0 | 100% | 131.9 ± 64.7 | 49.0% | 14 | 75.6% | 88% | 9.4 ± 8.7 | 92.1% | 64 | 96.4% | 5.5 × 10−37 |
2061.7505 | 4_4_2_1_0 | 22% | 0.3 ± 0.5 | 208.6% | 149 | 99.8% | 99% | 118.9 ± 45.6 | 38.4% | 17 | 74.2% | 4.7 × 10−34 |
4500.5968 | 7_7_5_2_2 | 2% | 0.0 ± 0.2 | 640.3% | 299 | 100.0% | 98% | 14.9 ± 6.3 | 42.0% | 56 | 95.5% | 1.1 × 10−30 |
1566.5553 | 4_3_0_1_0 | 100% | 218.8 ± 37.8 | 17.3% | 8 | 65.2% | 100% | 115.1 ± 36.1 | 31.3% | 19 | 76.5% | 2.0 × 10−30 |
2078.7658 | 5_4_3_0_0 | 46% | 2.5 ± 4.0 | 159.8% | 79 | 99.3% | 100% | 20.3 ± 7.2 | 35.3% | 53 | 95.0% | 5.1 × 10−30 |
2239.7982 | 6_4_1_1_0 | 100% | 81.3 ± 35.9 | 44.1% | 29 | 90.2% | 100% | 217.1 ± 56.0 | 25.8% | 11 | 65.7% | 2.0 × 10−28 |
1275.4599 | 4_3_0_0_0 | 100% | 105.6 ± 18.3 | 17.4% | 18 | 79.9% | 100% | 60.7 ± 17.6 | 29.0% | 31 | 87.1% | 2.4 × 10−26 |
1712.6132 | 4_3_1_1_0 | 100% | 25.6 ± 9.0 | 35.3% | 40 | 95.7% | 97% | 11.4 ± 5.6 | 49.6% | 60 | 96.0% | 2.0 × 10−20 |
1964.6977 | 7_4_0_0_0 | 95% | 7.1 ± 4.5 | 63.6% | 63 | 98.6% | 100% | 22.8 ± 8.8 | 38.8% | 50 | 94.4% | 1.2 × 10−19 |
2824.0047 | 7_6_1_0_1 | 93% | 6.5 ± 2.3 | 36.1% | 66 | 98.8% | 55% | 2.1 ± 2.2 | 105.8% | 108 | 98.1% | 4.3 × 10−19 |
2996.1147 | 6_7_4_0_0 | 2% | 0.0 ± 0.1 | 640.3% | 303 | 100.0% | 89% | 3.3 ± 2.1 | 62.4% | 86 | 97.5% | 3.1 × 10−18 |
1259.4650 | 3_3_1_0_0 | 100% | 559.1 ± 72.7 | 13.0% | 5 | 55.6% | 100% | 374.7 ± 111.6 | 29.8% | 5 | 49.0% | 4.4 × 10−17 |
2093.7403 | 6_4_0_1_0 | 100% | 198.5 ± 57.3 | 28.9% | 9 | 67.2% | 100% | 321.4 ± 71.9 | 22.4% | 7 | 55.6% | 5.7 × 10−17 |
1437.5127 | 5_3_0_0_0 | 100% | 27.4 ± 9.4 | 34.2% | 39 | 95.4% | 100% | 53.3 ± 16.7 | 31.4% | 35 | 89.3% | 5.3 × 10−16 |
2077.7454 | 5_4_1_1_0 | 100% | 198.3 ± 63.2 | 31.9% | 10 | 69.1% | 100% | 356.6 ± 103.0 | 28.9% | 6 | 52.4% | 1.3 × 10−15 |
1582.5502 | 4_3_0_0_1 | 100% | 14.9 ± 7.3 | 49.1% | 49 | 97.3% | 95% | 6.1 ± 3.9 | 63.9% | 72 | 97.0% | 2.3 × 10−15 |
1599.5655 | 6_3_0_0_0 | 100% | 8.3 ± 3.6 | 43.1% | 57 | 98.2% | 100% | 19.3 ± 7.5 | 38.7% | 54 | 95.2% | 2.6 × 10−15 |
3901.3907 | 7_7_3_2_1 | 17% | 0.3 ± 0.9 | 271.5% | 139 | 99.8% | 93% | 11.5 ± 8.1 | 69.9% | 58 | 95.8% | 4.7 × 10−15 |
1745.6234 | 6_3_1_0_0 | 100% | 12.0 ± 7.9 | 65.6% | 52 | 97.7% | 100% | 36.2 ± 16.6 | 45.9% | 42 | 92.2% | 4.8 × 10−15 |
1421.5178 | 4_3_1_0_0 | 100% | 184.4 ± 45.2 | 24.5% | 12 | 72.8% | 100% | 118.0 ± 39.0 | 33.0% | 18 | 75.4% | 1.7 × 10−14 |
2036.7188 | 6_3_1_1_0 | 90% | 6.7 ± 4.2 | 62.7% | 65 | 98.8% | 100% | 22.4 ± 11.3 | 50.6% | 51 | 94.6% | 1.9 × 10−14 |
1462.5444 | 3_4_1_0_0 | 100% | 1301.9 ± 137.8 | 10.6% | 2 | 28.6% | 100% | 984.8 ± 219.8 | 22.3% | 2 | 28.9% | 3.5 × 10−14 |
1973.7345 | 4_5_2_0_0 | 5% | 0.0 ± 0.2 | 530.9% | 264 | 100.0% | 96% | 9.0 ± 6.8 | 75.8% | 65 | 96.5% | 6.7 × 10−14 |
1947.6824 | 5_4_0_0_1 | 100% | 84.7 ± 36.7 | 43.3% | 26 | 87.7% | 100% | 160.8 ± 54.0 | 33.6% | 13 | 68.9% | 1.8 × 10−13 |
2255.7931 | 7_4_0_1_0 | 100% | 52.0 ± 25.5 | 49.2% | 35 | 93.9% | 100% | 110.1 ± 45.9 | 41.6% | 21 | 78.6% | 5.0 × 10−12 |
2109.7352 | 6_4_0_0_1 | 98% | 17.0 ± 10.9 | 64.0% | 47 | 97.0% | 100% | 31.8 ± 11.3 | 35.4% | 46 | 93.5% | 8.7 × 10−11 |
Category | m/z | N-glycan | Class a | AUC | P/C Ratio b | p-Value | |
---|---|---|---|---|---|---|---|
(A) | Major | 910.3277 | 3_2_0_0_0 | CORE | 1.000 | −3.0 | 2.7 × 10−66 |
1056.3856 | 3_2_1_0_0 | C/HB-F | 1.000 | −3.1 | 1.4 × 10−60 | ||
2061.7505 | 4_4_2_1_0 | C-FS | 0.993 | 461.6 | 4.7 × 10−34 | ||
2296.8197 | 6_5_0_1_0 | C-S | 0.991 | −14.0 | 5.5 × 10−37 | ||
2368.8408 | 5_4_1_2_0 | C-FS | 0.985 | −3.2 | 1.5 × 10−37 | ||
Minor | 1754.6602 | 3_4_3_0_0 | C-F | 1.000 | A/P | 1.0 × 10−40 | |
1916.7130 | 4_4_3_0_0 | C-F | 0.998 | 28.6 | 8.0 × 10−38 | ||
4500.5968 | 7_7_5_2_2 | C-FS | 0.989 | 603.7 | 1.1 × 10−30 | ||
2078.7658 | 5_4_3_0_0 | C/HB-F | 0.981 | 8.2 | 5.1 × 10−30 | ||
1973.7345 | 4_5_2_0_0 | C-F | 0.975 | 234.7 | 6.7 × 10−14 | ||
3901.3907 | 7_7_3_2_1 | C-FS | 0.954 | 36.1 | 4.7 × 10−15 | ||
2996.1147 | 6_7_4_0_0 | C-FS | 0.944 | 146.8 | 3.1 × 10−18 | ||
1964.6977 | 7_4_0_0_0 | HB | 0.940 | 3.2 | 1.2 × 10−19 | ||
3698.3113 | 7_6_3_2_1 | C-FS | 0.924 | A/P | 1.6 × 10−13 | ||
2824.0047 | 7_6_1_0_1 | C-FS | 0.920 | −3.1 | 4.3 × 10−19 | ||
2036.7188 | 6_3_1_1_0 | HB-FS | 0.906 | 3.4 | 1.9 × 10−14 | ||
(B) | Major | 2239.7982 | 6_4_1_1_0 | HB-FS | 0.973 | 2.7 | 2.0 × 10−28 |
1566.5553 | 4_3_0_1_0 | C/HB-S | 0.969 | −1.9 | 2.0 × 10−30 | ||
1275.4599 | 4_3_0_0_0 | C/HB | 0.955 | −1.7 | 2.4 × 10−26 | ||
1259.4650 | 3_3_1_0_0 | C/HB-F | 0.921 | −1.5 | 4.4 × 10−17 | ||
1437.5127 | 5_3_0_0_0 | HB | 0.921 | 1.9 | 5.3 × 10−16 | ||
2093.7403 | 6_4_0_1_0 | HB-S | 0.911 | 1.6 | 5.7 × 10−17 | ||
2077.7454 | 5_4_1_1_0 | C/HB-FS | 0.909 | 1.8 | 1.3 × 10−15 | ||
1462.5444 | 3_4_1_0_0 | C-F | 0.900 | −1.3 | 3.5 × 10−14 | ||
Minor | 1599.5655 | 6_3_0_0_0 | HB | 0.921 | 2.3 | 2.6 × 10−15 |
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Lee, H.; Lee, A.; Seo, N.; Oh, J.; Kweon, O.-K.; An, H.J.; Kim, J. Discovery of N-glycan Biomarkers for the Canine Osteoarthritis. Life 2020, 10, 199. https://doi.org/10.3390/life10090199
Lee H, Lee A, Seo N, Oh J, Kweon O-K, An HJ, Kim J. Discovery of N-glycan Biomarkers for the Canine Osteoarthritis. Life. 2020; 10(9):199. https://doi.org/10.3390/life10090199
Chicago/Turabian StyleLee, Hyunjun, Ahyun Lee, Nari Seo, Jiwon Oh, Oh-Kyeong Kweon, Hyun Joo An, and Jaehan Kim. 2020. "Discovery of N-glycan Biomarkers for the Canine Osteoarthritis" Life 10, no. 9: 199. https://doi.org/10.3390/life10090199