Publications by authors named "Andreas Ruppert"

4 Publications

  • Page 1 of 1

Adenovirus Particle Quantification in Cell Lysates Using Light Scattering.

Hum Gene Ther Methods 2017 10 14;28(5):268-276. Epub 2017 Aug 14.

1 Klinikum Rechts der Isar, Technische Universität München , Klinik und Poliklinik für Urologie, München, Germany.

Adenoviral vector production for therapeutic applications is a well-established routine process. However, current methods for measurement of adenovirus particle titers as a quality characteristic require highly purified virus preparations. While purified virus is typically obtained in the last step of downstream purification, rapid and reliable methods for adenovirus particle quantification in intermediate products and crude lysates to allow for optimization and validation of cell cultures and intermediate downstream processing steps are currently not at hand. Light scattering is an established process to measure virus particles' size, though due to cell impurities, adequate quantification of adenovirus particles in cell lysates by light scattering has been impossible until today. This report describes a new method using light scattering to measure virus concentration in nonpurified cell lysates. Here we report application of light scattering, a routine method to measure virus particle size, to virus quantification in enzymatically conditioned crude lysates. Samples are incubated with phospholipase A2 and benzonase and filtered through a 0.22 μm filter cartridge prior to quantification by light scattering. Our results show that this treatment provides a precise method for fast and easy determination of total adenovirus particle numbers in cell lysates and is useful to monitor virus recovery throughout all downstream processing.
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http://dx.doi.org/10.1089/hgtb.2017.052DOI Listing
October 2017

A genome-wide association study in chronic obstructive pulmonary disease (COPD): identification of two major susceptibility loci.

PLoS Genet 2009 Mar 20;5(3):e1000421. Epub 2009 Mar 20.

Genetics Division, GlaxoSmithKline Research and Development, Research Triangle Park, NC, USA.

There is considerable variability in the susceptibility of smokers to develop chronic obstructive pulmonary disease (COPD). The only known genetic risk factor is severe deficiency of alpha(1)-antitrypsin, which is present in 1-2% of individuals with COPD. We conducted a genome-wide association study (GWAS) in a homogenous case-control cohort from Bergen, Norway (823 COPD cases and 810 smoking controls) and evaluated the top 100 single nucleotide polymorphisms (SNPs) in the family-based International COPD Genetics Network (ICGN; 1891 Caucasian individuals from 606 pedigrees) study. The polymorphisms that showed replication were further evaluated in 389 subjects from the US National Emphysema Treatment Trial (NETT) and 472 controls from the Normative Aging Study (NAS) and then in a fourth cohort of 949 individuals from 127 extended pedigrees from the Boston Early-Onset COPD population. Logistic regression models with adjustments of covariates were used to analyze the case-control populations. Family-based association analyses were conducted for a diagnosis of COPD and lung function in the family populations. Two SNPs at the alpha-nicotinic acetylcholine receptor (CHRNA 3/5) locus were identified in the genome-wide association study. They showed unambiguous replication in the ICGN family-based analysis and in the NETT case-control analysis with combined p-values of 1.48 x 10(-10), (rs8034191) and 5.74 x 10(-10) (rs1051730). Furthermore, these SNPs were significantly associated with lung function in both the ICGN and Boston Early-Onset COPD populations. The C allele of the rs8034191 SNP was estimated to have a population attributable risk for COPD of 12.2%. The association of hedgehog interacting protein (HHIP) locus on chromosome 4 was also consistently replicated, but did not reach genome-wide significance levels. Genome-wide significant association of the HHIP locus with lung function was identified in the Framingham Heart study (Wilk et al., companion article in this issue of PLoS Genetics; doi:10.1371/journal.pgen.1000429). The CHRNA 3/5 and the HHIP loci make a significant contribution to the risk of COPD. CHRNA3/5 is the same locus that has been implicated in the risk of lung cancer.
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http://dx.doi.org/10.1371/journal.pgen.1000421DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2650282PMC
March 2009

A genome-wide investigation of SNPs and CNVs in schizophrenia.

PLoS Genet 2009 Feb 6;5(2):e1000373. Epub 2009 Feb 6.

Institute for Genome Sciences and Policy, Duke University, Durham, North Carolina, USA.

We report a genome-wide assessment of single nucleotide polymorphisms (SNPs) and copy number variants (CNVs) in schizophrenia. We investigated SNPs using 871 patients and 863 controls, following up the top hits in four independent cohorts comprising 1,460 patients and 12,995 controls, all of European origin. We found no genome-wide significant associations, nor could we provide support for any previously reported candidate gene or genome-wide associations. We went on to examine CNVs using a subset of 1,013 cases and 1,084 controls of European ancestry, and a further set of 60 cases and 64 controls of African ancestry. We found that eight cases and zero controls carried deletions greater than 2 Mb, of which two, at 8p22 and 16p13.11-p12.4, are newly reported here. A further evaluation of 1,378 controls identified no deletions greater than 2 Mb, suggesting a high prior probability of disease involvement when such deletions are observed in cases. We also provide further evidence for some smaller, previously reported, schizophrenia-associated CNVs, such as those in NRXN1 and APBA2. We could not provide strong support for the hypothesis that schizophrenia patients have a significantly greater "load" of large (>100 kb), rare CNVs, nor could we find common CNVs that associate with schizophrenia. Finally, we did not provide support for the suggestion that schizophrenia-associated CNVs may preferentially disrupt genes in neurodevelopmental pathways. Collectively, these analyses provide the first integrated study of SNPs and CNVs in schizophrenia and support the emerging view that rare deleterious variants may be more important in schizophrenia predisposition than common polymorphisms. While our analyses do not suggest that implicated CNVs impinge on particular key pathways, we do support the contribution of specific genomic regions in schizophrenia, presumably due to recurrent mutation. On balance, these data suggest that very few schizophrenia patients share identical genomic causation, potentially complicating efforts to personalize treatment regimens.
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http://dx.doi.org/10.1371/journal.pgen.1000373DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2631150PMC
February 2009

Thermodynamic and structural characterization of the transformation from a metastable low-density to a very high-density form of supercooled TIP4P-Ew model water.

Chemphyschem 2008 Dec;9(18):2737-41

Physikalische Chemie, Fakultät Chemie, TU Dortmund, Otto-Hahn-Str. 6,D-44227 Dortmund, Germany.

We explore the phase diagram of the metastable TIP4P-Ew liquid model water from 360 K down to 150 K at densities ranging from 0.950 to 1.355 g cm(-3). In addition to the low-density/high-density (LDL/HDL) liquid-liquid transition, we observe a structural high-density/very high-density (HDL/VHDL) transformation for the lowest temperatures at 1.30 g cm(-3). The characteristics of the isobars and isotherms suggest the presence of a stepwise HDL/VHDL transition with first-order-like appearance. In addition, we also identify an apparent pretransition at 1.24 g cm(-3), which suggests that the experimentally detected LDA/VHDA transformation might evolve into a multiple-step process with different local structures representing local minima in the free-energy landscape. Such a scenario is supported by a pronounced correlation between the isothermal density dependence of the pressure, with a stepwise increase of the oxygen coordination number, due to the appearance of interstitial water molecules.
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http://dx.doi.org/10.1002/cphc.200800539DOI Listing
December 2008