Development of a serum free medium for HUMIRA^® biosimilar by design of experiment approaches

Abstract

BackgroundSerum have been traditionally used to support growth of animal cell cultures. However, the increasing growth of therapeutic biopharmaceuticals market, accelerated the high demand for the serum-free medium (SFM).ObjectiveThe main objective is to design a SFM for a stable rCHO cell line that produces a fully anti-human TNF-α monoclonal antibody (mAb) corresponding to HUMIRA® biosimilar.Materials and methodsDesign of Experiment (DoE) approaches were used to determine the key factors due to their effect on specific growth rate and mAb production. The production was carried out in T-flasks at different initial cell concentrations and then in Erlenmeyers with the developed SFM. mAb production was compared with commercial SFMs in terms of yield and productivity.ResultsRegarding to our findings, when the developed SFM-adapted cells were compared with the cells produced in commercial SFMs, the mAb productivity in developed SFM were higher (1.3–1.6 times) depending on higher mAb concentration and less (3–5 times) cell concentration. Additionally, the produced mAb in the developed SFM provided high conformational similarity with its originator HUMIRA®.ConclusionDoE approaches could be used to reduce cost and time in designing SFM for any commercially important cell line to produce high value biologics.

A study of the THP‐1 cell line as the potential biologics production platform with the emphasis on serum‐free media substitution for economic expediency

“…The cells’ growth kinetics were measured using hemocytometer cell counts with trypan blue from daily samples, and Equations (1) and (2) were used to calculate the maximum specific growth rate ( μ max ) and doubling time ( t d ), respectively. [ 36 ] $$\begin{equation} {\mu }_{{\rm{max}}} = \frac{{\ln \left( {{x}_2} \right) - {\rm{ln}}\left( {{x}_1} \right)}}{{{t}_2 - {t}_1}} \end{equation}$$ $$\begin{equation} {t}_{\mathrm{d}}=\frac{\ln \left(2\right)}{\mu} \end{equation}$$where x 1 is the cell concentration of the first day of the logarithmic phase, x 2 the cell concentration of the last day of the logarithmic phase, t 1 the first day of the logarithmic phase, and t 2 is the last day of logarithmic phase.…”

“…were used to calculate the maximum specific growth rate (μ max ) and doubling time (t d ), respectively. [36] 𝜇 max = ln (x 2 ) − ln (x 1 )…”

Serum-Free Medium for Recombinant Protein Expression in Chinese Hamster Ovary Cells

“…In addition, some special trace elements and lipid mixtures need to be further optimized, which requires high cost. Future studies should aim at reducing the overall cost by beginning with the optimization of these two categories of substances ( Raab et al, 2010 ; Xiao et al, 2014 ; Kimiz-Gebologlu et al, 2020 ). Among them, the type and culture mode of the bioreactor also affect the quality of the final product considerably.…”

Development of serum substitute medium for bone tissue engineering

“…), applications (cell-based therapy, food industry, or cell-seeded implants), and species (human, canine, etc.) [8][9][10][11][12][13][14][15][16][17][18]. These variations in medium formulations indicate that one universal serum substitute medium might not exist but it should be developed/optimized for each specific cell type and specific application.…”