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Figure S1: Ma et al., 2015 Supplementary Data 1: Southern blot of three individual plants per generation (T5, T6 and T7), following the digestion of genomic.

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Presentation on theme: "Figure S1: Ma et al., 2015 Supplementary Data 1: Southern blot of three individual plants per generation (T5, T6 and T7), following the digestion of genomic."— Presentation transcript:

1 Figure S1: Ma et al., 2015 Supplementary Data 1: Southern blot of three individual plants per generation (T5, T6 and T7), following the digestion of genomic DNA (10 mg per lane) with NcoI and XbaI. Coding sequences for antibody genes were used as the probe (left panel = heavy chain, right panel = light chain).

2 Figure S2: Ma et al., 2015 Supplementary Data 2: Northern blot of individual plants from three generations (three plants from T5, three from T6 and six plants from T7). Coding sequences for antibody genes were used as probe (left panel = heavy chain transcript, right panel = light chain transcript). RiboRuler High Range RNA ladder (Fermentas) was used as the MW marker.

3 Figure S3: Ma et al., 2015 Sample Major signals observed (Da)Possible assignment Calculated average chemical mass (Da) P2G12, batch 090918 (M-Scan No 1300) 148,6722 x LC + 2 x HC (without CTerm K) + 2 x Hex3HexNAc4FucPent 148,667 148,7972 x LC + 1 x HC (without CTerm K) + 1 x HC (with CTerm K)+ 2 x Hex3HexNAc4FucPent 148, 795 P2G12, batch 080902 (M-Scan No 1301) 148,6772 x LC + 2 x HC (without CTerm K) + 2 x Hex3HexNAc4FucPent 148,677 148,8012 x LC + 1 x HC (without CTerm K) + 1 x HC (with CTerm K)+ 2 x Hex3HexNAc4FucPent 148,795 Sample UV peak at RT (min) Major signals observed (Da) Possible assignmentsCalculated average chemical mass (Da) P2G12, batch 090918 (M-Scan No 1300) 21.823,3221 x LC23,325 23.151,0171 x HC (without C- Term K) + 1 x Hex3HexNAc4FucPent 51,024 23.151,1451 x HC + 1 x Hex3HexNAc4FucPent 51,152 P2G12, batch 080902 (M-Scan No 1301) 21.723,3211 x LC23,325 22.751,0161 x HC (without C- Term K) + 1 x Hex3HexNAc4FucPent 51,024 22.751,1441 x HC + 1 x Hex3HexNAc4FucPent 51,152 A B Supplementary Data 3: Calculated chemical mass by LC/ES-MS analysis after A) desalting; and B) reduction.

4 Figure S4: Ma et al., 2015 A B Supplementary Data 4: Peptide mapping obtained from online LC/ES-MS analysis of a reduced/carboxymethylated tryptic digest of sample batches. a) light chain polypeptide; and b) heavy chain polypeptide. Mapped sequences are shown in bold and underlined.

5 Figure S5: Ma et al., 2015 Monosaccharide P2G12, clinical batch 090918 P2G12, pre-clinical batch 080902 Mean nmoles/mg 1 Ratio relative to mannose = 3.0 2 Mean nmoles/mg 1 Ratio relative to mannose = 3.0 2 Fucose 210.916 1.0 Xylose 291.322 1.4 Mannose 693.048 3.0 Galactose 1.80.0781.2 0.075 Glucose 2.30.101.9 0.12 GalNAc ND/ / GlcNAc 572.539 2.4 Supplementary Data 5: Monosaccharide analysis of P2G12.

6 Suppl. Data 6: Ma et al., 2015 Figure S6: Map of the plant expression vector pTRAp-2G12-Ds. Open reading frames are shown as arrows. Sequences transferred to the plant genome are shown in green. (LB/RB – left and right border of T-DNA from A. tumefaciens Ti-plasmid); Pnos – nopaline synthase gene promoter; pat – Streptomyces hygroscopicus gene for phosphinothricin acetyltransferase; pAnos – nopaline synthase gene polyadenylation signal; SAR – scaffold attachment region of the N. tabacum RB7 gene; P35SS – CaMV 35S promoter with duplicated transcriptional enhancer; TL – 5' untranslated region from Tobacco etch virus; pA35S – CaMV 35S polyadenylation signal; 2G12Hc – 2G12 heavy chain gene, including native signal peptide (SPg); 2G12LC - 2G12 light chain gene, including native signal peptide (SPk); TP Hordeum vulgare GBSSI transit peptide sequence; DsRed – Discosoma spp. red fluorescent protein gene; RK2 ori – origin of replication; bla – b-lactamase gene; ColE1ori – origin of replication.

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