Susana Alonso Villela

In the biopharmaceutical industry, nanobodies show promising properties for its application in serotherapy for serums targeting the most toxic components of scorpion venom. Nevertheless, the limited number of scientific publications on the production of nanobodies for antivenom productions poses a challenge. The reported yield is low (< 4 mg/L), biomass production is not reported, and the production in high cell density cultures have not been studied. Production conditions (temperature of ...

5L bioreactor culture of strain E. coli WK6 in a batch/fed-batch strategy at 37°C. Characterization of biomass growth and oxygen transfer

5L bioreactor culture of recombinant strain E. coli CH10-12 in a batch/fed-batch strategy at 37°C (BR02). Protein induction promoted when biomass reached 25 g cdw/L and at 28°C for 12 h

5L bioreactor culture of recombinant strain E. coli CH10-12. Protein induction promoter when biomass reached 30 g cdw/L and at 29°C for 38 h

5L bioreactor culture of reference strain E. coli WK6. Protein induction promoter when biomass reached 25 g cdw/L and at 29°C for 28 h

5L bioreactor culture of recombinant strain E. coli CH10-12. Protein induction promoter when biomass reached 25 g cdw/L and at 30°C for 10 h

Samples taken in the 5L bioreactor BR01 in the culture of E. coli WK6 in batch and fed-batch strategy

Gas analyzer (INNOVA 1313, Lumasense Technologies) Channel 3: gas out - BR02 Channel 4: gas out - BR03 Channel 6: gas in (reference) Sampling time: 60 s Flushig time: 45 s Sampling time: 15 s

Gas analyzer (INNOVA 1313, Lumasense Technologies) Channel 4: gas out - BR01 Channel 6: gas in (reference) Sampling time: 60 s Flushing time: 45 s Analysis time: 15 s

5L culture of CH10-12, 37°C batch/fed-batch culture with induction at 30°C for 10h

5L culture of CH10-12, 37°C batch/fed-batch culture with induction at 30°C for 10h

Nanobody production phase, IPTG 1 mM added to the bioreactor

Biomass production phase

Batch phase in 5L bioreactor culture of E coli CH10-12

5L bioreactor culture of strain E. coli WK6 at 37°C. Characterization of biomass growth and oxygen transfer in the fed-batch phase

5L bioreactor culture of strain E. coli WK6 at 37°C. Characterization of biomass growth and oxygen transfer in the batch phase

Procedure followed for the data treatment and analysis of 5L bioreactor cultures

Sample taken from the 5L bioreactors, cell dry weight and HPLC data

Gas analyzer (INNOVA 1313, Lumasense Technologies) Channel 3: gas out - BR04 Channel 4: gas out - BR05 Channel 6: gas in (reference) Sampling time: 60 s Flushig time: 45 s Sampling time: 15 s

Gas analyzer (INNOVA 1313, Lumasense Technologies) Channel 3: gas out - BR02 Channel 4: gas out - BR03 Channel 6: gas in (reference) Sampling time: 60 s Flushig time: 45 s Sampling time: 15 s

5L bioreactor of E. coli NbF12-10 in a batch/fed-batch strategy at 37°C, induction promoted for 37h at 29°C MFCS BioPAT Sartorius

5L bioreactor of E. coli WK6 in a batch/fed-batch strategy at 37°C, induction promoted for 28h at 29°C MFCS BioPAT Sartorius

5L bioreactor of E. coli CH10-12 in a batch/fed-batch strategy at 37°C, induction promoted for 35h at 32°C MFCS BioPAT Sartorius