TransIT-PRO Transfection Kit은 부유 세포인 293 cell과 CHO cell에서 포유류 단백질을 생산하도록
개발된 DNA transfection 시약과 boost 시약으로 구성되어 있다. PRO Boost Reagent는 배지에 따라
선택적으로 사용하는 시약으로 발현을 증가시키는 역할을 한다. TransIT-PRO Transfection Reagent와
PRO Boost Reagent는 animal-origin free components로 구성되어 있으며 화학적으로 규정된 다양한 배지에
적용 가능하다. TransIT-PRO Transfection Kit은 transfection후의 배지 교환이 필요하지 않고
transient transfection이나 stable transfection 모두에 적용 가능하다.
High Performance: A Cost-effective Alternative for Protein Production
Figure 1. Achieve High Antibody Titers Using the TransIT-PRO Transfection Kit in Suspension CHO Cells.
Human IgG1 was produced by transient transfection using TransIT-PRO and PRO Boost Reagent (1:1:1),
25 kDa linear PEI (6:1) or FreeStyle MAX (1:1) transfection reagents according to the manufacturers or published
protocol (reagent:DNA ratio). Transfections were performed using 1 μg plasmid DNA per milliliter of culture
and 0.5 x 106 cells/ml at the time of transfection. FreeStyle™ CHO-S cells were cultured in 20 ml of
FreeStyle™ CHO Expression medium in 125 ml shake flasks. (A) Day 3, 5 and 7 supernatants were clarified
and analyzed using a human IgG-Fc sandwich ELISA. Error bars represent the standard deviation of triplicate technical replicates,
25kDa linear PEI is duplicate technical replicates. (B) Day 7 supernatants were clarified and analyzed by Western blot.
An IgG standard was included for quantification estimate (S1= 1.6 mg/L, S2= 3.2 mg/L, S3 = 6.3 mg/L).
Easy to Use: Compatible with Multiple Media Formulations
Figure 2. TransIT-PRO™ Provides High Performance Across Varied Media Formulations.
FreeStyle CHO-S cells were adapted to five representative growth media including: BD Select™ CD1000 medium
(Becton, Dickinson and Company, Franklin Lakes, NJ), BD Select™ CHO medium (Becton, Dickinson and Company, Franklin Lakes, NJ),
FreeStyle™ CHO Expression medium (Life Technologies Corporation, Carlsbad, CA),
ProCHO5 medium (Lonza Inc., Allendale, NJ) and PowerCHO2 CD medium (Lonza Inc., Allendale, NJ).
Cells were transfected with a plasmid using the TransIT-PRO™ and PRO Boost Reagent (1:1:1),
25 kDa linear PEI (4:1) (Polysciences, Warrington, PA), or FreeStyle™ MAX (1:1) (Life Technologies Corporation, Carlsbad, CA)
transfection reagents according to the manufacturers or published protocol (reagent:DNA ratio).
Transfections were performed in 24-well deep well shaker blocks using 1 μg plasmid DNA per milliliter of culture
and 0.5 x 106 cells/ml at the time of transfection. (A) Luciferase expression was assessed 48 hours
post-transfection using a conventional luciferase assay. Error bars represent the standard deviation of duplicate wells.
(B) Human IgG1 was quantitated from day 5 clarified supernatants and analyzed by a human anti-Fc sandwich ELISA.
Error bars represent the standard deviation of triplicate wells.
Scale-up:
Figure 3. Scaling of Transient Transfection Using TransIT-PRO Transfection Kit is Linear From 4 to 400 Milliliters.
Human IgG1 was produced by transient transfection using the TransIT-PRO Transfection Kit and a plasmid encoding
a human IgG1 construct. A DNA concentration was 1μg/ml of culture and a ratio of TransIT-PRO:PRO
Boost Reagent:DNA ratio of 1:0.5:1. Cells were plated at a density of 0.5 x 106 cells/ml at the time of transfection.
CHO-S cells were cultured in BD Select CD1000 media using 4 ml per well of a 24-well deep well shake block,
40 ml in 125 ml Erlenmeyer shake flask, 40 ml in 125 ml 2 sidearm spinner flask and 400 ml in 500 ml 2 sidearm spinner flask.
Day 3, 5 and 7 supernatants were clarified and analyzed by an anti-Fc sandwich ELISA. Error bars represent the
standard deviation of triplicate technical replicates.
