Bioreactors for high cell density and continuous multi-stage cultivations: options for process intensification in cell culture-based viral vaccine production

With an increasing demand for efficacious, safe, and affordable vaccines for human and animal use, process intensification in cell culture-based viral vaccine production demands advanced process strategies to overcome the limitations of conventional batch cultivations. However, the use of fed-batch, perfusion, or continuous modes to drive processes at high cell density (HCD) and overextended operating times has so far been little explored in large-scale viral vaccine manufacturing. Also, possible reductions in cell-specific virus yields for HCD cultivations have been reported frequently. Taking into account that vaccine production is one of the most heavily regulated industries in the pharmaceutical sector with tough margins to meet, it is understandable that process intensification is being considered by both academia and industry as a next step toward more efficient viral vaccine production processes only recently. Compared to conventional batch processes, fed-batch and perfusion strategies could result in ten to a hundred times higher product yields. Both cultivation strategies can be implemented to achieve cell concentrations exceeding 10(7) cells/mL or even 10(8) cells/mL, while keeping low levels of metabolites that potentially inhibit cell growth and virus replication. The trend towards HCD processes is supported by development of GMP-compliant cultivation platforms, i.e., acoustic settlers, hollow fiber bioreactors, and hollow fiber-based perfusion systems including tangential flow filtration (TFF) or alternating tangential flow (ATF) technologies. In this review, these process modes are discussed in detail and compared with conventional batch processes based on productivity indicators such as space-time yield, cell concentration, and product titers. In addition, options for the production of viral vaccines in continuous multi-stage bioreactors such as two- and three-stage systems are addressed. While such systems have shown similar virus titers compared to batch cultivations, keeping high yields for extended production times is still a challenge. Overall, we demonstrate that process intensification of cell culture-based viral vaccine production can be realized by the consequent application of fed-batch, perfusion, and continuous systems with a significant increase in productivity. The potential for even further improvements is high, considering recent developments in establishment of new (designer) cell lines, better characterization of host cell metabolism, advances in media design, and the use of mathematical models as a tool for process optimization and control

Near-infrared spectroscopy of stellar populations in nearby spiral galaxies

We present high spatial resolution, medium spectral resolution near-infrared (NIR) H- and K-band long-slit spectroscopy for a sample of 29 nearby (z < 0.01) inactive spiral galaxies, to study the composition of their NIR stellar populations. These spectra contain a wealth of diagnostic stellar absorption lines, e.g. MgI 1.575 micron, SiI 1.588 micron, CO (6-3) 1.619 micron, MgI 1.711 micron, NaI 2.207 micron, CaI 2.263 micron and the 12CO and 13CO bandheads longward of 2.29 micron. We use NIR absorption features to study the stellar population and star formation properties of the spiral galaxies along the Hubble sequence, and we produce the first high spatial resolution NIR HK-band template spectra for low redshift spiral galaxies along the Hubble sequence. These templates will find applications in a variety of galaxy studies. The strength of the absorption lines depends on the luminosity and/or temperature of stars and, therefore, spectral indices can be used to trace the stellar population of galaxies. The entire sample testifies that the evolved red stars completely dominate the NIR spectra, and that the hot young star contribution is virtually nonexistent.Comment: 13 pages, 5 figures. Accepted to MNRAS. arXiv admin note: text overlap with arXiv:astro-ph/040313

Search for CP Violation in the decays D+ -> K_S pi+ and D+ -> K_S K+

A high statistics sample of photo-produced charm from the FOCUS(E831) experiment at Fermilab has been used to search for direct CP violation in the decays D+->K_S pi+ and D+ -> K_S K+. We have measured the following asymmetry parameters relative to D+->K-pi+pi+: A_CP(K_S pi+) = (-1.6 +/- 1.5 +/- 0.9)%, A_CP(K_S K+) = (+6.9 +/- 6.0 +/- 1.5)% and A_CP(K_S K+) = (+7.1 +/- 6.1 +/- 1.2)% relative to D+->K_S pi+. The first errors quoted are statistical and the second are systematic. We also measure the relative branching ratios: \Gamma(D+->\bar{K0}pi+)/\Gamma(D+->K-pi+pi+) = (30.60 +/- 0.46 +/- 0.32)%, \Gamma(D+->\bar{K0}K+)/\Gamma(D+->K-pi+pi+) = (6.04 +/- 0.35 +/- 0.30)% and \Gamma(D+->\bar{K0}K+)/\Gamma(D+->\bar{K0}pi+) = (19.96 +/- 1.19 +/- 0.96)%.Comment: 4 pages, 3 figure

Search for Λc+→pK+π−\Lambda_c^+ \to p K^+ \pi^- and Ds+→K+K+π−D_s^+ \to K^+ K^+ \pi^- Using Genetic Programming Event Selection

We apply a genetic programming technique to search for the double Cabibbo suppressed decays $\Lambda_c^+ \to p K^+ \pi^-$ and $D_s^+ \to K^+ K^+ \pi^-$. We normalize these decays to their Cabibbo favored partners and find $\text{BR}($\Lambda_c^+ \to p K^+ \pi^-$)/\text{BR}($\Lambda_c^+ \to p K^- \pi^+$) = (0.05 \pm 0.26 \pm 0.02)$ and $\text{BR}($D_s^+ \to K^+ K^+ \pi^-$)/\text{BR}($D_s^+ \to K^+ K^- \pi^+$) = (0.52\pm 0.17\pm 0.11)$ where the first errors are statistical and the second are systematic. Expressed as 90% confidence levels (CL), we find $< 0.46$ and $< 0.78$ respectively. This is the first successful use of genetic programming in a high energy physics data analysis.Comment: 10 page

A Non-parametric Approach to the D+ to K*0bar mu+ nu Form Factors

Using a large sample of D+ -> K- pi+ mu+ nu decays collected by the FOCUS photoproduction experiment at Fermilab, we present the first measurements of the helicity basis form factors free from the assumption of spectroscopic pole dominance. We also present the first information on the form factor that controls the s-wave interference discussed in a previous paper by the FOCUS collaboration. We find reasonable agreement with the usual assumption of spectroscopic pole dominance and measured form factor ratios.Comment: 14 pages, 5 figures, and 2 tables. We updated the previous version by changing some words, removing one plot, and adding two tables. These changes are mostly stylisti

Measurement of the relative branching ratio BR(\Xi_c^+ \to p^+ K^-\pi^+)\BR(\Xi_c^+ \to \Xi^- \pi^+ \pi^+)

We report the observation of the Cabibbo suppressed decay \Xi_c^+ \to p K^-\pi^+ using data collected with the FOCUS spectrometer during the 1996--97 Fermilab fixed target run. We find a \Xi_c^+ signal peak of 202\pm35 events. We have measured the relative branching ratios BR(\Xi^+_c\to p K^-\pi^+)/BR(\Xi^+_c\to\Xi^-\pi^+\pi^+)= 0.234 \pm 0.047 \pm 0.022 and BR(\Xi^+_c\to p \bar{K}^*(892)^0)/BR(\Xi^+_c\to p K^-\pi^+)= 0.54 \pm 0.09 \pm 0.05 .Comment: 9 pages, 4 figure

Measurements of Ξc+\Xi_c^{+} Branching Ratios

Using data collected by the fixed target Fermilab experiment FOCUS, we measure the branching ratios of the Cabibbo favored decays $\Xi_c^+ \to \Sigma^+K^-\pi^+$, $\Xi_c^+ \to \Sigma^+ \bar{K}^{*}(892)^0$, and $\Xi_c^+ \to \Lambda^0K^-\pi^+\pi^+$ relative to $\Xi_c^+ \to \Xi^-\pi^+\pi^+$ to be $0.91\pm0.11\pm0.04$, $0.78\pm0.16\pm0.06$, and $0.28\pm0.06\pm0.06$, respectively. We report the first observation of the Cabibbo suppressed decay $\Xi_c^+ \to \Sigma^+K^+K^-$ and we measure the branching ratio relative to $\Xi_c^+ \to \Sigma^+K^-\pi^+$ to be $0.16\pm0.06\pm0.01$. We also set 90% confidence level upper limits for $\Xi_c^+ \to \Sigma^+ \phi$ and $\Xi_c^+ \to \Xi^*(1690)^0(\Sigma^+ K^-) K^+$ relative to $\Xi_c^+ \to \Sigma^+K^-\pi^+$ to be 0.12 and 0.05, respectively. We find an indication of the decays $\Xi_c^+ \to \Omega^-K^{+}\pi^+$ and $\Xi_c^+ \to \Sigma^{*}(1385)^+ \bar{K}^0$ and set 90% confidence level upper limits for the branching ratios with respect to $\Xi_c^+ \to \Xi^-\pi^+\pi^+$ to be 0.12 and 1.72, respectively. Finally, we determine the 90% C.L. upper limit for the resonant contribution $\Xi_c^+ \to \Xi^{*}(1530)^0 \pi^+$ relative to $\Xi_c^+ \to \Xi^-\pi^+\pi^+$ to be 0.10.Comment: 14 pages, 8 figure

A Measurement of the Ds+ Lifetime

A high statistics measurement of the Ds+ lifetime from the Fermilab fixed-target FOCUS photoproduction experiment is presented. We describe the analysis of the two decay modes, Ds+ -> phi(1020)pi+ and Ds+ -> \bar{K}*(892)0K+, used for the measurement. The measured lifetime is 507.4 +/- 5.5 (stat.) +/- 5.1 (syst.) fs using 8961 +/- 105 Ds+ -> phi(1020)pi+ and 4680 +/- 90 Ds+ -> \bar{K}*(892)0K+ decays. This is a significant improvement over the present world average.Comment: 5 pages, 3 figures, 2 tables, submitted to PR

A Non-parametric Approach to Measuring the \kpi{} Amplitudes in \dpkkpi{} Decay

Using a large sample of \dpkkpi{} decays collected by the FOCUS photoproduction experiment at Fermilab, we present the first non-parametric analysis of the \kpi{} amplitudes in \dpkkpi{} decay. The technique is similar to the technique used for our non-parametric measurements of the \krzmndk{} form factors. Although these results are in rough agreement with those of E687, we observe a wider S-wave contribution for the \ksw{} contribution than the standard, PDG \cite{pdg} Breit-Wigner parameterization. We have some weaker evidence for the existence of a new, D-wave component at low values of the $K^- \pi^+$ mass.Comment: 13 pages 3 figure

Dalitz plot analysis of D_s+ and D+ decay to pi+pi-pi+ using the K-matrix formalism

FOCUS results from Dalitz plot analysis of D_s+ and D+ to pi+pi-pi+ are presented. The K-matrix formalism is applied to charm decays for the first time to fully exploit the already existing knowledge coming from the light-meson spectroscopy experiments. In particular all the measured dynamics of the S-wave pipi scattering, characterized by broad/overlapping resonances and large non-resonant background, can be properly included. This paper studies the extent to which the K-matrix approach is able to reproduce the observed Dalitz plot and thus help us to understand the underlying dynamics. The results are discussed, along with their possible implications on the controversial nature of the sigma meson.Comment: To be submitted to Phys.Lett.B A misprint corrected in formula