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.

The Wrong Kind of Lean: Over-Commitment and Under-Represented Skills on Technology Teams

This paper reports on results from five companies in the aerospace and automotive industries to show that over-commitment of technical professionals and under-representation of key skills on technology development and transition teams seriously impairs team performance. The research finds that 40 percent of the projects studied were inadequately staffed, resulting in weaker team communications and alignment. Most importantly, the weak staffing on these teams is found to be associated with a doubling of project failure rate to reach full production. Those weakly staffed teams that did successfully insert technology into production systems were also much more likely than other teams to have development delays and late engineering changes. The conclusion suggests that the expense of project failure, delay and late engineering changes in these companies must greatly out-weigh the savings gained from reduced staffing costs, and that this problem is likely going to be found in other technology-intensive firms intent on seeing project budgets as a cost to be minimized rather than an investment to be maximized.

Computer Recognition of Prismatic Solids

An investigation is made into the problem of constructing a model of the appearance to an optical input device of scenes consisting of plane-faced geometric solids. The goal is to study algorithms which find the real straight edges in the scenes, taking into account smooth variations in intensity over faces of the solids, blurring of edges and noise. A general mathematical analysis is made of optimal methods for identifying the edge lines in figures, given a raster of intensities covering the entire field of view. There is given in addition a suboptimal statistical decision procedure, based on the model, for the identification of a line within a narrow band on the field of view given an array of intensities from within the band. A computer program has been written and extensively tested which implements this procedure and extracts lines from real scenes. Other programs were written which judge the completeness of extracted sets of lines, and propose and test for additional lines which had escaped initial detection. The performance of these programs is discussed in relation to the theory derived from the model, and with regard to their use of global information in detecting and proposing lines.

Prototype of a Configurable Web-Based Assessment System

The MIT Prototype Educational Assessment System provides subjects and courses at MIT with the ability to perform online assessment. The system includes polices to handle harassment and electronic "flaming" while protecting privacy. Within these frameworks, individual courses and subjects can make their own policy decisions about such matters as to when assessments can occur, who can submit assessments, and how anonymous assessments are. By allowing assessment to take place continually and allowing both students and staff to participate, the system can provide a forum for the online discussion of subjects. Even in the case of scheduled assessments, the system can provide advantages over end-of-term assessment, since the scheduled assessments can occur several times during the semester, allowing subjects to identify and adjust those areas that could use improvement. Subjects can also develop customized questionnaires, perhaps in response to previous assessments, to suit their needs.