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Table 1 Pre-induction yields of hA 2a R are maintained at pilot scale even in the presence of cytotoxic methanol concentrations

From: Functional recombinant protein is present in the pre-induction phases of Pichia pastoris cultures when grown in bioreactors, but not shake-flasks

  Phase Age (h) Residual glycerol (g L-1) Residual methanol (g L-1) DCW (g L-1) Total membrane protein yield in 1 L culture (mg) Bmaxestimate (pmol mg-1) Total hA2aR yield in 1 L culture (pmol) Specific yield (pmol g-1)
Low methanol cultivation IIIA 42.2 0 (0) 0 (0) 41.91 (0.6) 764.4 4.4 (0.1) 3363.6 80.3
IIIB 44.4 1.1 (0.2) 0.5 (0.1) 42.10 (0.4) 2791.1 7.1 (0.6) 19816.9 470.7
IV 66.4 0 (0) 1.0 (0.1) 50.53 (0.5) 4084.4 4.5 (0.2) 18380.0 363.7
IV 89.0 0 (0) 0 (0) 71.82 (0.2) 7704.4 5.0 (0.2) 38522.2 536.4
High methanol cultivation IIIA 42.2 2.9 (0.1) 0 (0) 46.71 (0.5) 813.3 4.4 (0.2) 3578.7 76.6
IIIB 44.4 4.2 (0.2) 2.1 (0.2) 46.66 (0.7) 3060.0 4.0 (0.0) 12240.0 262.3
IV 66.4 1.1 (0.2) 39.1 (9.0) 45.76 (0.5) 3277.8 3.8 (0.1) 12455.5 272.2
V 88.9 0 (0) 84.7 (0.5) 37.64 (0.8) 1797.8 3.1 (0.1) 5573.1 148.1
  1. The total yield of hA2aR (pmol) and the specific yield of hA2aR per gram of DCW (pmol g-1) were derived from the Bmax and total membrane protein measurements, as described in the Methods section. Values are calculated per L of culture for the transition and induction phases of the low and high methanol cultivations and are the mean of triplicate determinations, with the standard error of the mean in parentheses. Triplicate measurements were from three different membrane preparations and standard error of the mean is shown in parentheses.