Automated Analysis report for PHA-E Vector CFG

Array Chart

Figure 1. Barcharts of raw glycan binding data. Glycans are grouped by family (automatically if not provided) and sorted by binding strength. Mouseover to see glycan structures.

Primary Motifs

Figure 2. Boxplots of glycans grouped by primary motif. Individual points are glycans. Mouseover to see glycan structures.
Primary Motif ID Motif Graphic Structure Relative Binding Number of Glycans
A A0p 0.84 59
B B0p 1.00 3
C C0p 0.63 11
0 0.00 536
Figure 2. Boxplots of glycans grouped by primary motif. Individual points are glycans. Mouseover to see glycan structures.
Primary Motif ID Motif Graphic Structure Relative Binding Number of Glycans
A A0p 0.70 59
B B0p 1.00 3
C C0p 0.20 11
0 0.00 536
Figure 2. Boxplots of glycans grouped by primary motif. Individual points are glycans. Mouseover to see glycan structures.
Primary Motif ID Motif Graphic Structure Relative Binding Number of Glycans
A A0p 1.00 59
B B0p 0.87 3
C C0p 0.12 11
0 0.00 536
Figure 2. Boxplots of glycans grouped by primary motif. Individual points are glycans. Mouseover to see glycan structures.
Primary Motif ID Motif Graphic Structure Relative Binding Number of Glycans
A A0p 1.00 59
B B0p 0.56 3
C C0p 0.08 11
0 0.00 536
Figure 2. Boxplots of glycans grouped by primary motif. Individual points are glycans. Mouseover to see glycan structures.
Primary Motif ID Motif Graphic Structure Kd Number of Glycans
A A0p 114.32 59
B B0p 6.1127 3
C C0p 104.043 11
0 392.51 17

All Motifs

Figure 3. Boxplots of glycans grouped by motif. Individual points are glycans. Mouseover to see glycan structures.
Motif ID Nearest Common Name (Accuracy%**) Motif Graphic Structure Relative Binding Number of Glycans P-Value***
A3 Type 2 DiLacNAc Biantennary N-Glycan (98%) A3p 1.00 18 0.000
A5 Bisecting N-Glycan (97%) A5p 0.97 2 0.000
A1 Triantennary 6' N-Glycan (99%) A1p 0.75 11 0.000
A0* Bisecting N-Glycan (97%) A0p 0.72 14 0.000
A4 Biantennary N-Glycan Terminal Type 2 LacNAc (97%) A4p 0.68 8 0.000
A2 Type 2 DiLacNAc 5-Acetyl Neuramic Acid (98%) A2p 0.24 6 0.000
B0 Terminal N-Glycan N-Acetyl Glucosamine (98%) B0p 0.92 3 0.000
C0 Biantennary N-Glycan Terminal Hexose (94%) C0p 0.58 11 0.000
0 Non-Binders (100%) 0.00 536 NA
Figure 3. Boxplots of glycans grouped by motif. Individual points are glycans. Mouseover to see glycan structures.
Motif ID Nearest Common Name (Accuracy%**) Motif Graphic Structure Relative Binding Number of Glycans P-Value***
A5 Bisecting N-Glycan (97%) A5p 1.00 2 0.000
A3 Type 2 DiLacNAc Biantennary N-Glycan (98%) A3p 0.91 18 0.000
A0* Bisecting N-Glycan (97%) A0p 0.53 14 0.000
A4 Biantennary N-Glycan Terminal Type 2 LacNAc (97%) A4p 0.47 8 0.000
A1 Triantennary 6' N-Glycan (99%) A1p 0.36 11 0.000
A2 Type 2 DiLacNAc 5-Acetyl Neuramic Acid (98%) A2p 0.10 6 0.037
B0 Terminal N-Glycan N-Acetyl Glucosamine (98%) B0p 0.82 3 0.000
C0 Biantennary N-Glycan Terminal Hexose (94%) C0p 0.17 11 0.000
0 Non-Binders (100%) 0.00 536 NA
Figure 3. Boxplots of glycans grouped by motif. Individual points are glycans. Mouseover to see glycan structures.
Motif ID Nearest Common Name (Accuracy%**) Motif Graphic Structure Relative Binding Number of Glycans P-Value***
A5 Bisecting N-Glycan (97%) A5p 1.00 2 0.000
A3 Type 2 DiLacNAc Biantennary N-Glycan (98%) A3p 0.46 18 0.000
A0* Bisecting N-Glycan (97%) A0p 0.30 14 0.000
A4 Biantennary N-Glycan Terminal Type 2 LacNAc (97%) A4p 0.20 8 0.000
A1 Triantennary 6' N-Glycan (99%) A1p 0.14 11 0.000
A2 Type 2 DiLacNAc 5-Acetyl Neuramic Acid (98%) A2p 0.03 6 0.990
B0 Terminal N-Glycan N-Acetyl Glucosamine (98%) B0p 0.26 3 0.000
C0 Biantennary N-Glycan Terminal Hexose (94%) C0p 0.04 11 0.729
0 Non-Binders (100%) 0.00 536 NA
Figure 3. Boxplots of glycans grouped by motif. Individual points are glycans. Mouseover to see glycan structures.
Motif ID Nearest Common Name (Accuracy%**) Motif Graphic Structure Relative Binding Number of Glycans P-Value***
A5 Bisecting N-Glycan (97%) A5p 1.00 2 0.000
A3 Type 2 DiLacNAc Biantennary N-Glycan (98%) A3p 0.50 18 0.000
A0* Bisecting N-Glycan (97%) A0p 0.27 14 0.000
A4 Biantennary N-Glycan Terminal Type 2 LacNAc (97%) A4p 0.13 8 0.001
A2 Type 2 DiLacNAc 5-Acetyl Neuramic Acid (98%) A2p 0.10 6 0.118
A1 Triantennary 6' N-Glycan (99%) A1p 0.08 11 0.046
B0 Terminal N-Glycan N-Acetyl Glucosamine (98%) B0p 0.16 3 0.027
C0 Biantennary N-Glycan Terminal Hexose (94%) C0p 0.02 11 0.989
0 Non-Binders (100%) 0.00 536 NA
Figure 3. Boxplots of glycans grouped by motif. Individual points are glycans. Mouseover to see glycan structures.
Motif ID Nearest Common Name (Accuracy%**) Motif Graphic Structure Kd Number of Glycans P-Value***
A5 Bisecting N-Glycan (97%) A5p 1.50424 2 0.257
A3 Type 2 DiLacNAc Biantennary N-Glycan (98%) A3p 7.22808 18 0.000
A1 Triantennary 6' N-Glycan (99%) A1p 32.3941 11 0.004
A0* Bisecting N-Glycan (97%) A0p 84.6645 14 0.009
A4 Biantennary N-Glycan Terminal Type 2 LacNAc (97%) A4p 139.606 8 0.149
A2 Type 2 DiLacNAc 5-Acetyl Neuramic Acid (98%) A2p 658.882 6 0.190
B0 Terminal N-Glycan N-Acetyl Glucosamine (98%) B0p 6.1127 3 0.117
C0 Biantennary N-Glycan Terminal Hexose (94%) C0p 104.043 11 0.032
0 Non-Binders (100%) 392.51 17 NA

*Motif indicates the remaining glycans not matched by motifs which are a subset. Motif definition needs to be taken in the context of the model.

**Accuracy describes the consistency between common-name definition of the motif and the formal, text-based definition of the motif, in terms of percent agreement in the glycans containing the two motifs. Common Name label definitions given here.

***P-Value refers to difference from Non-Binders with multiple testing correction (Dunnet’s Test)

Motifs with a red motif ID fail to show a logistic response to protein concentration in the range of concentrations analyzed. These motifs may be nonbinding motifs (motifs which define nonbinding exceptions) or simply fail to fit. Nonbinding motifs are determined based on concentration dependent response when available or the average binding of non-motif glycans otherwise.

Motif Intensity Maps

Figure 4. Glycan intensity and motif distribution plot. The top half of the plot presents the observed glycan binding intensity of various glycans used in the array over their rank binding intensity; only the top glycans are shown. The second plot indicates the position of glycans containing the various motifs in the top plot with a yellow tick.

Treemap Diagram

*Motif indicates the remaining glycans not matched by motifs which are a subset. Motif definition needs to be taken in the context of the model.

Figure 5. Treemap of glycan binding grouped by motif and family structure. The model structure can be represented as nested boxes where box size is proportional to the number of glycans with the motif and color changes with change in average relative binding of glycans with the motif. Only three layers of data splitting are included here, though further splitting may be possible.

Model Diagram

*Motif matches the remaining glycans not matched by earlier motifs in the model.

Figure 6. Tree representation of the regression tree model trained on array data. Data flows through the tree (top-down) and is split by the various motifs. The motif used the split the data at each point has the id “family+split number” except when further split. In the case of futher splits the id of the motif used to split the data is denoted with an asterisk.

Binding Curves

Figure 7. Logistic curves fit to average motif binding and glycan binding. Curves are fit with as many parameters as possible given the data. All curves are based on the logistic curve with a fixed intercept of 0. Mouse-over to view curve info and highlight glycans/motifs on the other plot.

Figure 8. Scatterplots of logistic curve parameters for glycans (points) and motifs (text).

Motif Details

Motif ID Motif Graphic   Motif Full Graphic Kd Asymptote Slope Motif Text
A1 A1p A1Full 28.204 40005 0.867 {^x{<2f4f6f>GalB1-4<3f6f>GlcNAcB1-2(<3f6f>GlcNAcB1-6)<3f4f>ManA1-6(<2f4f6f>GalB1-4<3f6f>GlcNAcB1-2<3f4f6f>ManA1-3)<2f4f>ManB1-4<3f6f>GlcNAcB1-4<3f>GlcNAcB}N}
A2 A2p A2Full 43.956 14557 0.870 {{<3f6f8f><GlcNAcorNeu5Ac>?<1or2>-<3or6><2f4f>GalB1-4<3f6f>GlcNAcB1-3<2f4f6f>GalB1-4<3f6f>GlcNAcB1-2<3f4f6f>ManA1-6(<3f6f8f><GlcNAcorNeu5Ac>?<1or2>-<3or6><2f4f>GalB1-4<3f6f>GlcNAcB1-3<2f4f6f>GalB1-4<3f6f>GlcNAcB1-2<3f4f6f>ManA1-3)<2f4f>ManB1-4<3f6f>GlcNAcB1-4<3f6f>GlcNAcB}N} {Spacer:Asn}
A3 A3p A3Full 4.414 41224 1.025 {VA{<3f6f8f><GlcNAcorNeu5Ac>?<1or2>-3<2f4f6f>GalB1-4<3f6f>GlcNAcB1-2<3f4f6f>ManA1-6(<3f6f8f><GlcNAcorNeu5Ac>?<1or2>-3<2f4f6f>GalB1-4<3f6f>GlcNAcB1-2<3f4f6f>ManA1-3)<2f4f>ManB1-4<3f6f>GlcNAcB1-4<3f>GlcNAcB}NK}
A4 A4p A4Full 9.623 29799 0.953 {{<2f4f6f>GalB1-4<3f6f>GlcNAcB1-2<3f4f6f>ManA1-6(<2f4f6f>GalB1-4<3f6f>GlcNAcB1-2<3f4f6f>ManA1-3)<2f4f>ManB1-4<3f6f>GlcNAcB1-4<3f>GlcNAcB}N}
A5 A5p A5Full 1.225 35813 1.166 {<4f6f8f>Neu5AcA2-3<2f4f6f>GalB1-4<3f6f>GlcNAcB1-<2or4><3f6f>ManA1-6(<3f4f6f>GlcNAcB1-4)<2f>ManB}
A0 A0p A0Full 9.001 32521 0.831 {<2f4f6f>GalB1-4<3f6f>GlcNAcB1-<2or4><3f>ManA1-6(<2f4f6f>GalB1-4<3f6f>GlcNAcB1-2<3f6f>ManA1-3)<2f>ManB}
B0 B0p B0Full 5.078 37225 1.262 {<3f4f6f>GlcNAcB1-2<3f4f6f>ManA1-6(<3f4f6f>GlcNAcB1-4)<2f>ManB}
C0 C0p C0Full 53.062 34950 1.032 {<6f>GalB1-4<3f6f>GlcNAcB1-2<3f4f6f>ManA1-<3or6>(<3f6f>GlcNAcB1-2<3f4f>ManA1-<3or6>)<2f4f>Man?1-4<3f6f>GlcNAcB1-4<3f6f>GlcNAcB}

See Symbol Nomenclature for Glycans (SNFG) for complete key: https://www.ncbi.nlm.nih.gov/glycans/snfg.html