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Table 3 Growth and rhA1AT productivities of top 2 cell pools from each vector set #

From: Engineering selection stringency on expression vector for the production of recombinant human alpha1-antitrypsin using Chinese Hamster ovary cells

MTX concentration Vector Pool 1 Pool 2 Average
Max titer % (mg/l) q p (pcd) t D (h) Fold titer increase + Max titer % (mg/l) q p (pcd) t D (h) Fold titer increase + Max titer % (mg/l) q p (pcd) t D (h)
50 nM pAID 256 17.6 46 5.6      256 17.6 46
pAIDp 492 25.7 36 4.2 244 10.9 36 3.1 368 18.3 36
pAIDpp 647 29.0 33 4.4 539 25.9 44 5.4 593 27.5 39
pAI772Dp 1054 41.3 28 8.5 937 33.8 26 8.2 996 37.6 27
pAID* 275 14.4 41 3.1 170 7.8 40 2.2 222 11.1 40
pAID*p 277 8.4 33 2.0      277 8.4 33
300 nM pAID 560 32.5 34 2.2      560 32.5 34
pAIDp 514 25.8 33 1.0 240 13.1 37 1.0 377 19.5 35
pAIDpp 1146 88.2 45 1.8 846 41.9 41 1.6 996 65.0 43
pAI772Dp 1111 48.6 42 1.1 863 35.4 27 0.9 987 42.0 34
pAID* 721 34.3 43 2.6 398 22.1 33 2.3 559 28.2 38
pAID*p 412 17.7 29 1.5      412 17.7 29
  1. #The 2 cell pools that gave the highest titers in 96 well-plate adherent culture with 50 nM MTX were chosen for further amplification to 300 nM MTX. The 50 nM and 300 nM MTX cell pools were then adapted to serum-free suspension culture in shake flasks to evaluate their growth and rhA1AT production profiles. The top producing cell pool in 96-well plate format (Figure 2) is indicated as Pool 1. Only 1 cell pool from vectors pAID and pAID*p survived the adaptation to suspension culture or the MTX amplification process respectively.
  2. %Maximum titer was taken as the highest rhA1AT titer assayed from the first 11 days of the batch culture.
  3. +Fold titer increase for cell pools in 50 nM MTX suspension culture was compared against the titers of the same cell pools in adherent 96 well plate cultures, whereas that for cell pools in 300 nM MTX suspension culture was compared against the titers of the same cell pools in 50 nM MTX suspension culture.