High-throughput screening approach for pro-cess optimization. Is disposable equipment also rentable in the small-scale segment?

To transfer the existing process to an ambr® 250, scale-down approaches including volumetric power input variation, gassing strategy and feeding volumes have been evaluated in triplicates. Growth and production were compared to controls (Fig. 2). Interestingly, the ambr® 250 profi les were closer to shake fl ask approaches than to the 10 L bioreactors. The process could not be completely mimicked. However, the most promising setting was further used.

A DoE was conducted in duplicates in ambr® (Fig. 3 A, B). After evaluation, modifi ed process parameters were transferred to a 2.7 L bench top system (Fig. 3 C, D). Although the absoluteresults in both systems (peak cell density and fi nal titer) were diff erent, the outcome of the ambr® screening could be verifi ed. The most promising setup was successfully scaled up to 10 L(Fig. 3 C, D: black lines).It can be clearly seen, that the process time could be reduced by 5 days without decreasing titer.

The disposable automated bioreactor system ambr® 250 promises good control and high capacitywith a short turnaround:

The in-house platform process (TurboCellTM platform) for the production of an IgG antibodyis used as a starting point:Cultivations of the TurboCellTM in diff erent scales and vessels show an up to 21 day long process with comparable cell growth (Fig. 1 A) and production behavior (Fig. 1 B): peak cell densities of 10 – 12∙106 cells per mL and up to 2.5 g product per L. The process represents a classical and successful scale-up starting from glass bioreactors.

•���The scale-down to ambr® 250 needs further adjustment

•���Nevertheless, the DoE could be successfully conducted in the single-use system and the platform process was improved

•���The effi ciency compared to our current stan-dard (10 L glass bioreactors) is outstanding (Fig. 4)

•���A decrease in labor input, but especially vast time saving within a multi-run project (due to 24 parallel runs), was shown

•���In terms of costs, the price of a disposable bioreactor plays a major role. However, the calculation for bench top bioreactors does not include abrasion of multi-use probes.

Figure 1: Cultivations of the TurboCellTM in lab-scale, pilot scale and production scale (P/V = const.); (A) viable cell concentration and viability, (B) product concentration

Figure 2: Cultivations of the TurboCellTM in ambr® 250; scale-down by adjustment of diff erent parameters in triplicates (similar colours); (A) viable cell concentration and viability ambr® 250, (B) product concentration all systems, (C) viable cell concentration and viability reference; (B10 = 10 L glass bioreactor, SF250 = 250 mL shake fl ask)

Figure 3: Cultivations of the TurboCellTM; setups include modifi cation of temperature, feeding strategy and cell seeding density; control shown in dark green (ambr9/ambr24/2,7 L glass bioreactor); (A) viable cell concentration and viability ambr® 250, (B) product concentration ambr® 250, (C) viable cell concentration and viability bench top, (D) product concentration bench top

Figure 4: Benefi ts of disposable system, compared to bench top systems (parameters normalized to the current standard: 10 glass bioreactors)

Single-use or multi-use – how proper does it work in small-scale?Stefan Wieschalka, Janine Seidemann, Anja Schäfer, Stefan R. SchmidtRentschler Biotechnologie GmbH, Laupheim Germany, stefan.wieschalka@rentschler.de

Single-use pros and cons Aim: process-time reduction, stable titer, fast approach

A case study

Conclusion

www.rentschler.de

•���Low investment (fast amortization)•���Small footprint (no piping)•���Simple & quick expansion (fl exibility)•���Short implementation time•���Reduced qualifi cation & maintenance•���Elimination of cleaning (QA/QC)•���Fast product change over (capacity)•���No cross-contamination risk (safety)

•���Continuous processing•���Orphan drug manufacturing•���Scale suitable for clinical supply

•���Scalability (reactors, columns, TFF)•���Transport sensitivity (columns)•���Increased waste•���Repetitive consumable costs•���Long supply lead times •���Storage requirements •���Limited standardization

Duration

Cost

Labor input10 L Glass bioreactor

2.7 L Glass bioreactor

ambr® 250

•���Supplier dependency•���Extractables & leachables eff ect

Time/(d)

Time/(d)

Time/(d)

Viab

le c

ell c

once

ntra

tion/

(106

mL-

1 )

Viab

ility/

(%)

Prod

uct c

once

ntra

tion/

(mg

L-1 )

Viab

le c

ell c

once

ntra

tion/

(106

mL-

1 )

Fina

l pro

duct

con

cent

ratio

n/(m

g L-

1 )

Cel

l cou

nt (1

06 m

L-1 )

Cel

l cou

nt (1

06 m

L-1 )

Cell viability (%

)

Time from`inoculation´

ambr1ambr6ambr11ambr16ambr21

ambr2ambr7ambr12ambr17ambr22

ambr3ambr8ambr13ambr18ambr23

ambr4ambr9ambr14ambr19ambr24

ambr5ambr10ambr15ambr20

ambr

1am

br2

ambr

3am

br4

ambr

5am

br6

ambr

7am

br8

ambr

9am

br10

ambr

11am

br12

ambr

13am

br14

ambr

15am

br16

ambr

17am

br18

ambr

19am

br20

ambr

21am

br22

ambr

23am

br24 B1

0SF

250_

1SF

250_

2

Cell viability (%

)

A

A

A

A

B

B C

Viab

ility/

(%)

Time from`inoculation´

Fina

l pro

duct

con

cent

ratio

n (m

g L-

1 )

C D

Viab

le c

ell c

once

ntra

tion/

(106

mL-

1 )

Viab

ility/

(%)

Prod

uct c

once

ntra

tion/

(mg

L-1 )

Time/(d)Time/(d)