An Extension to the Tactical Planning Model for a Job Shop: Continuous-Time Control

We develop an extension to the tactical planning model (TPM) for a job shop by the third author. The TPM is a discrete-time model in which all transitions occur at the start of each time period. The time period must be defined appropriately in order for the model to be meaningful. Each period must be short enough so that a job is unlikely to travel through more than one station in one period. At the same time, the time period needs to be long enough to justify the assumptions of continuous workflow and Markovian job movements. We build an extension to the TPM that overcomes this restriction of period sizing by permitting production control over shorter time intervals. We achieve this by deriving a continuous-time linear control rule for a single station. We then determine the first two moments of the production level and queue length for the workstation.

Over Expression of the CMP-sialic Acid Transporter in Chinese Hamster Ovary Cells Leads to Increased Sialylation

Most glyco-engineering approaches used to improve quality of recombinant glycoproteins involve the manipulation of glycosyltransferase and/or glycosidase expression. We investigated whether the over expression of nucleotide sugar transporters, particularly the CMP-sialic acid transporter (CMP-SAT), would be a means to improve the sialylation process in CHO cells. We hypothesized that increasing the expression of the CMP-SAT in the cells would increase the transport of the CMP-sialic acid in the Golgi lumen, hence increasing the intra-lumenal CMP-sialic acid pool, and resulting in a possible increase in sialylation extent of proteins being produced. We report the construction of a CMP-SAT expression vector which was used for transfection into CHO-IFNγ, a CHO cell line producing human IFNγ. This resulted in approximately 2 to 5 times increase in total CMP-SAT expression in some of the positive clones as compared to untransfected CHO-IFNγ, as determined using real-time PCR analysis. This in turn concurred with a 9.6% to 16.3% percent increase in site sialylation. This engineering approach has thus been identified as a novel means of improving sialylation in recombinant glycoprotein therapeutics. This strategy can be utilized feasibly on its own, or in combination with existing sialylation improvement strategies. It is believed that such multi-prong approaches are required to effectively manipulate the complex sialylation process, so as to bring us closer to the goal of producing recombinant glycoproteins of high and consistent sialylation from mammalian cells.