Advancing Raman model calibration for perfusion bioprocesses using spiked harvest libraries.

BACKGROUND: Raman spectroscopy has gained popularity to monitor multiple process indicators simultaneously in biopharmaceutical processes. However, robust and specific model calibration remains a challenge due to insufficient analyte variability to train the models and high cross-correlation of various media components and artifacts throughout the process. MAIN METHODS: A systematic Raman calibration workflow for perfusion processes enabling highly specific and fast model calibration was developed. Harvest libraries consisting of frozen harvest samples from multiple CHO cell culture bioreactors collected at different process times were established. Model calibration was subsequently performed in an offline setup using a flow cell by spiking process harvest with glucose, raffinose, galactose, mannose, and fructose. MAJOR RESULTS: In a screening phase, Raman spectroscopy was proven capable not only to distinguish sugars with similar chemical structures in perfusion harvest but also to quantify them independently in process-relevant concentrations. In a second phase, a robust and highly specific calibration model for simultaneous glucose (root mean square error prediction [RMSEP] = 0.32 g L-1 ) and raffinose (RMSEP = 0.17 g L-1 ) real-time monitoring was generated and verified in a third phase during a perfusion process. IMPLICATION: The proposed novel offline calibration workflow allowed proper Raman peak decoupling, reduced calibration time from months down to days, and can be applied to other analytes of interest including lactate, ammonia, amino acids, or product titer.

Pressure-Induced Phase Transitions in GeTe-Rich Ge-Sb-Te Alloys across the Rhombohedral-to-Cubic Transitions.

We demonstrate that pressure-induced amorphization in Ge-Sb-Te alloys across the ferroelectric-paraelectric transition can be represented as a mixture of coherently distorted rhombohedral Ge8Sb2Te11 and randomly distorted cubic Ge4Sb2Te7 and high-temperature Ge8Sb2Te11 phases. While coherent distortion in Ge8Sb2Te11 does not prevent the crystalline state from collapsing into its amorphous counterpart in a similar manner to pure GeTe, the pressure-amorphized Ge8Sb2Te11 phase begins to revert to the crystalline cubic phase at ∼9 GPa in contrast to Ge4Sb2Te7, which remains amorphous under ambient conditions when gradually decompressed from 40 GPa. Moreover, experimentally, it was observed that pressure-induced amorphization in Ge8Sb2Te11 is a temperature-dependent process. Ge8Sb2Te11 transforms into the amorphous phase at ∼27.5 and 25.2 GPa at room temperature and 408 K, respectively, and completely amorphizes at 32 GPa at 408 K, while some crystalline texture could be seen until 38 GPa (the last measurement point) at room temperature. To understand the origins of the temperature dependence of the pressure-induced amorphization process, density functional theory calculations were performed for compositions along the (GeTe)x - (Sb2Te3)1-x tie line under large hydrostatic pressures. The calculated results agreed well with the experimental data.

Laser desorption time-of-flight mass spectrometry of atomic switch memory Ge2Sb2Te5 bulk materials and its thin films.

RATIONALE: Although the structure of atomic switch Ge2Sb2Te5 (GST) thin films is well established, the composition of the clusters formed in the plasma plume during pulsed-laser deposition (PLD) is not known. Laser Desorption Ionization Time-of-Flight Mass Spectrometry (LDI-TOF MS) is an effective method for the generation and study of clusters formed by laser ablation of various solids and thus for determining their structural fragments. METHODS: LDI of bulk or PLD-deposited GST thin layers and of various precursors (Ge, Sb, Te, and Ge-Te or Sb-Te mixtures) using a nitrogen laser (337 nm) was applied while the mass spectra were recorded in positive and negative ion modes using a TOF mass spectrometer equipped with a reflectron while the stoichiometry of the clusters formed was determined via isotopic envelope analysis. RESULTS: The singly negatively or positively charged clusters identified from the LDI of GST were Ge, Ge2, GeTe, Ge2Te, Ten (n = 1-3), GeTe2, Ge2Te2, GeTe3, SbTe2, Sb2Te, GeSbTe2, Sb3Te and the low abundance ternary GeSbTe3, while the LDI of germanium telluride yielded Gem Ten (+) clusters (m = 1-3, n = 1-3). Several minor Ge-H clusters were also observed for pure germanium and for germanium telluride. Sbn clusters (n = 1-3) and the formation of binary TeSb, TeSb2 and TeSb3 clusters were detected when Sb2Te3 was examined. CONCLUSIONS: This is the first report that elucidates the stoichiometry of Gem Sbn Tep clusters formed in plasma when bulk or nano-layers of GST material are ablated. The clusters were found to be fragments of the original structure. The results might facilitate the development of PLD technology for this memory phase-change material.

Laser ablation of AgSbS(2) and cluster analysis by time-of-flight mass spectrometry.

Thin films of AgSbS(2) are important for phase-change memory applications. This solid is deposited by various techniques, such as metal organic chemical vapour deposition or laser ablation deposition, and the structure of AgSbS(2)(s), as either amorphous or crystalline, is already well characterized. The pulsed laser ablation deposition (PLD) of solid AgSbS(2) is also used as a manufacturing process. However, the processes in plasma have not been well studied. We have studied the laser ablation of synthesized AgSbS(2)(s) using a nitrogen laser of 337 nm and the clusters formed in the laser plume were identified. The ablation leads to the formation of various single charged ternary Ag(p)Sb(q)S(r) clusters. Negatively charged AgSbS(4) (-), AgSb(2)S(3) (-), AgSb(2)S(4) (-), AgSb(2)S(5) (-) and positively charged ternary AgSbS(+), AgSb(2)S(+), AgSb(2)S(2) (+), AgSb(2)S(3) (+) clusters were identified. The formation of several singly charged Ag(+), Ag(2) (-), Ag(3) (-), Sb(3) (+), Sb(3) (-), S(8) (+) ions and binary Ag(p)S(r) clusters such as AgSb(2) (-), Ag(3)S(-), SbS(r) (-) (r = 1-5), Sb(2)S(-), Sb(2)S(2) (-), Sb(3)S(r) (-) (r = 1-4) and AgS(2) (+), SbS(+), SbS(2) (+), Sb(2)S(+), Sb(2)S(2) (+), Sb(3)S(r) (+) (r = 1-4), AgSb(2) (+) was also observed. The stoichiometry of the clusters was determined via isotopic envelope analysis and computer modeling. The relation of the composition of the clusters to the crystal structure of AgSbS(2) is discussed.