Defining process design space for monoclonal antibody cell culture

The concept of design space has been taking root as a foundation of in‐process control strategies for biopharmaceutical manufacturing processes. During mapping of the process design space, the multidimensional combination of operational variables is studied to quantify the impact on process performance in terms of productivity and product quality. An efficient methodology to map the design space for a monoclonal antibody cell culture process is described. A failure modes and effects analysis (FMEA) was used as the basis for the process characterization exercise. This was followed by an integrated study of the inoculum stage of the process which includes progressive shake flask and seed bioreactor steps. The operating conditions for the seed bioreactor were studied in an integrated fashion with the production bioreactor using a two stage design of experiments (DOE) methodology to enable optimization of operating conditions. A two level Resolution IV design was followed by a central composite design (CCD). These experiments enabled identification of the edge of failure and classification of the operational parameters as non‐key, key or critical. In addition, the models generated from the data provide further insight into balancing productivity of the cell culture process with product quality considerations. Finally, process and product‐related impurity clearance was evaluated by studies linking the upstream process with downstream purification. Production bioreactor parameters that directly influence antibody charge variants and glycosylation in CHO systems were identified. Biotechnol. Bioeng. 2010;106: 894–905. © 2010 Wiley Periodicals, Inc.     

Geochemical assessment of fluoride enriched groundwater and health implications from a part of Yavtmal District,India

Abstract

A comprehensive study was conducted from a semi-arid part of Yavtmal District, Maharashtra, India through combination approaches of geochemical modeling and its health consequences. The groundwater quality assessment shows that 55% of groundwater samples have the concentration of fluoride above the desirable limit. The high Na⁺/Ca⁺ ratio (>1.0) suggest the occurrence of cation exchange, which is further supported by Scholler’ chloro-alkaline indices. The geochemical modeling reveals that the existence of CaCO₃ precipitation and CaF₂ in groundwater. Simulation analysis indicates the dissolution of calcite, gypsum, and albite and precipitation of dolomite, fluorite, halite, and K-feldspar along with cation exchange as the main water–rock interactions influencing the groundwater chemistry. This is further significantly supported by pollution index of groundwater (PIG). PIG indicates about 18% of total samples fall in very high pollution zone, 3% in high pollution zone, 8% in moderate pollution zone, 24% in low pollution zone, and remaining (47%) express insignificant pollution. The 28% of subject studied have skeletal fluorosis varying from mild to severe type. In different pollution zones, the affected persons by dental fluorosis are varying from 15% to 41%. A proper monitoring and treatment are required for high fluoride water before its use for drinking and cooking.     

An in vitro system derived from Friend erythroleukemia cells to study messenger RNA stability

A system consisting of 40-80S messenger ribonucleoprotein particles (mRNP) from stationary Friend erythroleukemia (FEL) cells was used to investigate the stability of mRNA in vitro. The majority of mRNP mRNAs were found to be stable when incubated for periods of up to ninety minutes at 37 degrees. Nonetheless, many mRNAs are greatly reduced in abundance, including ones for eucaryotic elongation factor Tu (eEF-Tu) and the 73-78 kDa polypeptide commonly found in association with the poly(A) tails of mRNA. A divalent cation dependent ribonuclease (probably an endoribonuclease) could be washed off mRNP by treatment of the particles with 0.5M NaCl. The mRNAs contained in the resultant salt washed mRNPs, including eEF-Tu, were stable when incubated in vitro.