Development of a measles vaccine production process in MRC-5 cells grown on Cytodex1 microcarriers and in a stirred bioreactor

作者：Trabelsi Khaled; Majoul Samy; Rourou Samia; Kallel Hela^*

来源：Applied Microbiology and Biotechnology, 2012, 93(3): 1031-1040.

DOI：10.1007/s00253-011-3574-y

• MRC-5 cells
• Cytodex 1 microcarriers
• Stirred bioreactor
• AIK-C measles strain
• Measles vaccine

Measles vaccination remains the most efficient way to control the spread of the virus. This work focuses on the production of a measles vaccine using stirred conditions as an advanced option for process scale up. Non-porous Cytodex 1 microcarriers were used to support MRC-5 cell growth in suspension cultures. Virus replication was first optimized in spinner flasks, and the effects of various operational parameters were investigated. Cell infection with AIK-C measles strain at an MOI (multiplicity of infection) of 0.005, without glucose regulation and in M199 medium, resulted in a virus titer of 10(6.25) TCID50 (median tissue culture infective dose)/ml. To optimize the production process in a 7-l bioreactor, we carried out various perfused cultures using minimum essential medium (MEM)+5% FCS diluted with phosphate-buffered saline (PBS). We achieved a high cell density level (4.1x10(6) cells/ml) with an efficient use of the medium when MEM+5% FCS diluted with PBS at 25% was used during the cell amplification step. Optimization of measles production in MRC-5 cells grown on Cytodex 1 beads in a 7-l bioreactor showed that perfusion was the most efficient when compared to repeated-batch culture. Perfusion at a rate of 0.25 V (reactor volume)/day showed the highest specific productivity (1.6 IVP [infectious virus particle] cell(-1) day(-1)). Testing of several stabilizers containing pharmaceutically improved components such as sugars, amino acids, and charged ions showed that the formulation composed of sucrose and MgCl2, led to the maintenance of the infectivity of the AIK-C measles virus strain to a significant level, when stored at +28 degrees C, + 4 degrees C and -60 degrees C.

• 出版日期2012-2