Supplementary Material

Supplementary Table 1.Compositions of four types of plant cell culture medium: SH, BDS, B5 and MS

Medium components (mg/L) / SH / BDS / B5 / MS
Macronutrients
NH4NO3 / 320 / 1650
KNO3 / 2500 / 2530 / 2500 / 1900
CaCl2.2H2O / 151 / 440 / 150 / 440
MgSO4.7H2O / 195.4 / 250.6 / 250 / 370
KH2PO4 / 170
NH4H2PO4 / 300
(NH4)2SO4 / 134 / 134
NaH2PO4.H2O / 172 / 150
Micronutrients
KI / 1 / 0.75 / 0.75 / 0.83
H3BO3 / 5 / 3 / 3 / 6.2
MnSO4.H2O / 10 / 10 / 10 / 22.3
ZnSO4.7H2O / 1 / 2 / 2 / 8.6
Na2MoO4.2H2O / 0.1 / 0.25 / 0.25 / 0.25
CuSO4.5H2O / 0.2 / 0.04 / 0.025 / 0.025
CoCl2.6H2O / 0.1 / 0.025 / 0.025 / 0.025
Na2EDTA / 20 / 36.5 / 37.3 / 37.3
FeSO4.7H2O / 15 / 27.8 / 27.8 / 27.8
Vitamins
Inositol / 1000 / 1000 / 100 / 100
Glycine / 2 / 2
Thiamine HCl / 5 / 5 / 10 / 0.1
Pyridoxine HCl / 0.5 / 0.5 / 0.5
Nicotinic acid / 5 / 5 / 0.5

Supplementary Table 2.Secreted protein yields ofhuman growth hormone(hGH)and interferon α2b (IFNα2) expressed in BY-2 cells with a (SP)10 and/or (SP)20 tag. The transgenic BY-2 cells were cultured in either MS or SH medium for 10 days before the detection of the recombinant protein yields in culture media. Yields were determined from 5-10 cell lines per construct.

Constructs / Protein yield in MS medium (mg/L) / Protein yield in SH medium (mg/L)
hGH-(SP)10 / 0.9-1.3 / 16-35 (Xu et al. 2010)
IFNα2-(SP)10 / 0.3-0.9 / 17-28 (Xu et al. 2007)
IFNα2-(SP)20 / 0.2-1.1 / 23-27 (Xu et al. 2007)

References

Xu J, Okada S, Tan L, Goodrum KJ, Kopchick JJ, Kieliszewski MJ (2010) Human growth hormone expressed in tobacco cells as an arabinogalactan-protein fusion glycoprotein has a prolonged serum life. Transgenic Res 19:849-867

Xu J, Tan L, Goodrum KJ, Kieliszewski MJ (2007) High-yields and extended serum half-life of human interferon alpha2b expressed in tobacco cells as arabinogalactan-protein fusions. Biotechnol Bioeng 97:997-1008


Supplementary Figure 1.Anti-EGFP Western blotting detection of the (SP)32-EGFP transgene products accumulated insideBY-2 cells. The cells were grown in eight different medium (M1 to M8)created by mixing and matching of three major groups of (inorganic salts, vitamins and growth regulators) of SH and MS medium. The BY-2 cells were harvested after 10 days of culture. Partially glycosylated (SP)32-EGFP and the EGFP domain cleaved from the (SP)32-EGFP fusion are indicated by the grey arrow and white arrow, respectively.

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