TRIDENT: an infrared camera optimized for the detection of methanated substellar companions around nearby stars

A near-infrared (0.85-2.5 microns) camera in use at the Canada-France-Hawaii Telescope and at the 1.6m telescope of the Observatoire du Mont-Megantic is described. The camera is based on a Hawaii-1 1024x1024 HgCdTe array detector. Its main feature is to acquire three simultaneous images at three wavelengths (simultaneous differential imaging) across the methane absorption bandhead at 1.6 micron, enabling an accurate subtraction of the stellar point spread function (PSF) and the detection of faint close methanated companions. The instrument has no coronagraph and features a fast (1 MHz) data acquisition system without reset anomaly, yielding high observing efficiencies on bright stars. The performance of the instrument is described, and it is illustrated by CFHT images of the nearby star Ups And. TRIDENT can detect (3 sigma) a methanated companion with DeltaH=10 at 0.5 arcsec from the star in one hour of observing time. Non-common path aberrations between the three optical paths are the limiting factors preventing further PSF attenuation. Reference star subtraction and instrument rotation improve the detection limit by one order of magnitude.Comment: 8 pages, 6 figures, to appear in SPIE 486

Combining metabolic and process engineering strategies to improve recombinant glycoprotein production and quality

Increasing recombinant protein production while ensuring a high and consistent protein quality remains a challenge in mammalian cell culture process development. In this work, we combined a nutrient substitution approach with a metabolic engineering strategy that improves glucose utilization efficiency. This combination allowed us to tackle both lactate and ammonia accumulation and investigate on potential synergistic effects on protein production and quality. To this end, HEK293 cells overexpressing the pyruvate yeast carboxylase (PYC2) and their parental cells, both stably producing the therapeutic glycoprotein interferon \u3b12b (IFN\u3b12b), were cultured in media deprived of glutamine but containing chosen substitutes. Among the tested substitutes, pyruvate led to the best improvement in growth (integral of viable cell density) for both cell lines in batch cultures, whereas the culture of PYC2 cells without neither glutamine nor any substitute displayed surprisingly enhanced IFN\u3b12b production. The drastic reduction in both lactate and ammonia in the cultures translated into extended high viability conditions and an increase in recombinant protein titer by up to 47% for the parental cells and the PYC2 cells. Product characterization performed by surface plasmon resonance biosensing using Sambucus nigra (SNA) lectin revealed that the increase in yield was however accompanied by a reduction in the degree of sialylation of the product. Supplementing cultures with glycosylation precursors and a cofactor were effective at counterbalancing the lack of glutamine and allowed improvement in IFN\u3b12b quality as evaluated by lectin affinity. Our study provides a strategy to reconcile protein productivity and quality and highlights the advantages of PYC2-overexpressing cells in glutamine-free conditions.Peer reviewed: YesNRC publication: Ye