Stefan Muellner

Protagen AG · General Management

Background: Chronic inflammation is frequently observed on histological analysis of malignant and non-malignant prostate specimens. It is a suspected supporting factor for prostate diseases and their progression and a main cause of false positive PSA tests in cancer screening. We hypothesized that inflammation induces autoantibodies, which may be...

Clinical parameters are equally distributed between high and low inflammation patient cohorts. All samples were tested for statistically significant differences between the two sample groups. Clinical parameters including age, C-reactive protein, Gleason Score, prostate volume, PSA and free PSA (Table 1) are equally distributed amongst the high and...

Autoantibody marker antigens are differentially expressed in benign tissue and malignant lesions of the prostate. A Representative images for SPAST, STX18 and SPOP immunohistochemical double stainings with the basal cell marker p63. Left side shows benign lesions, right side represents invasive prostate carcinoma. The basal cell marker, which is ab...

List of autoantigens included in the planar protein microarray (Table "Protein Array") and in the Luminex bead protein array (Table "Luminex"). Autoantigen proteins included in the planar protein array or in the Luminex bead protein array are listed. The number of positive samples of indivual auoantigens for the low and the high inflammation cohort...

List of 165 significantly upregulated autoantibodies in serum samples of the screening cohort, ranked according to p-Value. (XLSX)

We have previously proposed that specific proteins may form insoluble aggregates as a response to an illness-specific proteostatic dysbalance in a subset of brains from individuals with mental illness, as is the case for other chronic brain conditions. So far, established risk factors DISC1 and dysbindin were seen to specifically aggregate in a sub...

As Escherichia coli (E. coli) is well defined with respect to its genome and metabolism, it is a favored host organism for recombinant protein production. However, many processes for recombinant protein production run under suboptimal conditions caused by wrong or incomplete information from an improper screening procedure, because appropriate on-l...

Allergy is an immunological disorder of the upper airways, lung, skin, and the gut with a growing prevalence over the last decades in Western countries. Atopy, the genetic predisposition for allergy, is strongly dependent on familial inheritance and environmental factors. These observations call for predictive markers of progression from atopy to a...

Stefan MüllnerUnternehmensgründungen in der Biotechnologie brauchen verständige Gründer, kenntnisreiche Investoren und unterstützende Persönlichkeiten.

Recent developments in proteomics technology offer new opportunities for clinical applications in hospital or specialized laboratories including the identification of novel biomarkers, monitoring of disease, detecting adverse effects of drugs, and environmental hazards. Advanced spectrometry technologies and the development of new protein array for...

The field of clinical proteomics offers opportunities to identify new disease biomarkers in body fluids, cells and tissues. These biomarkers can be used in clinical applications for diagnosis, stratification of patients for specific treatment, or therapy monitoring. New protein array formats and improved spectrometry technologies have brought these...

We describe a novel procedure for determination and validation of off-target activities of anti-cluster designation antigen identity 44 (CD44) variant 6 recombinant antibodies by combining two complementary technology platforms; namely UNIchip AV-400, containing a printed serial dilution of CD44 variant 6 and approximately 400 different human prote...

CD4(+)CD25(+)Foxp3(+) regulatory T cells (CD25(+) Treg cells) direct the maintenance of immunological self-tolerance by active suppression of autoaggressive T-cell populations. However, the molecules mediating the anergic state and regulatory function of CD25(+) Treg cells are still elusive. Using differential proteomics, we identified galectin-10,...

CD4+ T helper cells are the key player for control and regulation of adaptive immune responses. In a healthy immune system they protect the body against a variety of pathogens but CD4+ T cells are also involved in pathogenesis and perpetuation of diverse autoimmune diseases like chronic rheumatic inflammation. Consequently, analysis of T cell activ...

Protein Microarrays enable the analysis of multiple binding events in parallel. Based on patented UNIclone technology, we have established a reproducible protein biochip platform to determine specific binding profiles to multiple targets in a quantitative way. The UNIchip protein biochips provide a novel and unique tool for antibody characterizatio...

Proteome studies with small sample amounts are difficult to perform, especially when membrane proteins are the focus of interest. In our study a new method for the analysis of scarce membrane protein samples combining large gel 2-D-CTAB/SDS-PAGE with fluorescence dye saturation labelling (satDIGE) was developed, allowing a highly sensitive differen...

IntroductionMaterials and Methods Cells and Culture ConditionsIn Vitro Activity TestingAffinity ChromatographyElectrophoresis and Protein IdentificationBIAcore AnalysisSynthesis of Acyl Cyanides Methyl 5-cyano-5-oxopentanoateMethyl 6-cyano-6-oxohexanoateMethyl-5-cyano-3-methyl-5-oxopentanoateResultsDiscussion Cells and Culture ConditionsIn Vitro Ac...

Human Proteome Organization HUPO 2006, p.1167

Protein microarrays with immobilised proteins on their surface are new analytical tools to overcome the current limits with respect to sample volume and throughput. They have a great potential as well with respect to multiplexing of complex samples, as a research tool and in diagnostics. Based on recent advances in this technology, new applications...

The human genome has been sequenced and the challenges of understanding the function of the newly discovered genes have been addressed. High-throughput technologies such as DNA microarrays have been developed for the profiling of gene expression patterns in whole organisms or tissues. Protein arrays are emerging to follow DNA chips as possible scre...

Not much more than 15 years ago a handful of visionary scientists around the world suggested to sequence and analyze not only the human genome but also as many genomes as possible in order to compare DNA as well as to deduce protein sequences. By that means they expected to get an idea about the organization of life. However, after now having now s...

With the genome sequences of several organisms now in public databases, the scientific community has realized that it is time to prepare for the next step: the understanding of biological systems or systems biology. Whereas genes contain the information for life, the encoded proteins and RNAs fulfill nearly all the functions, from replication to re...

The isoxazole derivative Leflunomide (HWA 486) is a novel immunoregulatory and anti-inflammatory drug. Affinity chromatography was used to purify and identify Leflunomide binding proteins, which might play a role as potential cellular targets in the molecular mode of action. The Leflunomide derivative A 0273 was covalently coupled to a Fractogel® m...

The analysis of bile acids in biological matrices, e.g. gall bladder bile and serum, depends on accurate and reproducible sample pre-treatment techniques. The present study compares published and commonly used protocols with a special focus on the analysis of serum bile acids. Since adsorption of bile acids to serum proteins has a dramatic impact o...

Prp2p, Prp16p, Prp22p, and Prp43p are members of the DEAH-box family of ATP-dependent putative RNA helicases required for pre-mRNA splicing in Saccharomyces cerevisiae. Recently, mammalian homologues of Prp43p and Prp22p have been described, supporting the idea that splicing in yeast and man is phylogenetically conserved. In this study, we show tha...

An in vitro model featuring important inflammatory cellular states was established, based on the murine monocyte/macrophage cell line RAW 264.7. Macrophages are key players in chronic inflammation, and major parts of the biochemical reactions taking place in vivo, e.g., the production of proinflammatory cytokines, can be triggered in vitro by stimu...

The combination of two-dimensional electrophoresis (2-DE) and subsequent Western blot analysis with antibodies directed against common cellular proteins is a straightforward and reliable method to quickly generate fix points in a protein map. In order to assure high accuracy in the allocation of protein spots, two different replica blotting methods...

The results of the genome projects and the obvious problems with handling, storing and interpretation of generated data pose a lot of pressure on existing strategies in drug target discovery and development of new therapeutic concepts. Genomic data, i.e. DNA/RNA sequences identified by subtractive expression pattern analysis, are not enough per se...

At the end of September '96, the Club de Bruxelles held a conference ("The Future of Biotechnologies in Europe") to discuss the present development and the future of biotechnology in Europe. The meeting was intended to serve as a forum for the preparation of the 5th EU framework ("Inventing Tomorrow"), which is currently being set up under the lead...

In order to analyse bile acids in biological matrices, e.g. rat bile and human serum, high performance liquid chromatography (HPLC) was coupled to continuous-flow fast atom bombardment mass spectrometry (CFFAB-MS). A gradient elution system which had already proved to be well suited for the quantitative determination of conjugated bile acids in bil...

Glycosyl-phosphatidylinositol-anchored membrane proteins (GPI-proteins) are normally identified either by cleavage of the lipid anchor using (glycosyl)phosphatidylinositol-specific phospholipases C or D (GPI-PLs) or by metabolic labeling of the lipid moiety with specific building blocks. Therefore, methods for discrimination between transmembrane p...

Two proteins, in previous work labeled by affinity markers derived from taurocholic acid, were purified and partially sequenced. Antibodies were raised against purified proteins, and cross-reactions were carefully checked. The influence of these antibodies upon taurocholic acid import into vesicles from rat liver plasma membranes was measured, and...

Evaluation of the yield of expression of exogeneous protein in transformed Escherichia coli cells by means of one-dimensional SDS-PAGE often leads to overestimation and miscalculation. For example, it is possible that proteins of similar size comigrate and thus mask the overexpressed product band. Therefore, two-dimensional electrophoresis was used...

Cholic acid (1) has been coupled via ω-aminoalkoxy spacer at C-3 to chlorambucil (3), HR 780 (4) and oxaproline peptide (5). Drug conjugates 9, 11, 18 exhibit strong affinity to specific bile acid transport systems.

Methodology for the preparation of 7α-12α-dihydroxy-3β- (2) and 7α,12α-dihydroxy-3α-(2-hydroxyethoxy)-5β-cholanic acid (3) is described. Nucleophilic displacement of the 3-mesylate of unprotected cholic acid with ethylene glycol led to the 3β-isomer whereas the 3α-isomer was synthesized via the 7,12-diacetyl protected 3-allyl ether of methyl cholat...

In evaluating the purification of genetically engineered human insulin there is no plausible correlation between the yield of expression as determined by SDS-PAGE (taking into account all normally occurring losses during the several purification steps) and the actual yield, i.e., the final pure product. The aim of our work was to develop a fast, ac...

For the purification of the bile acid transport proteins from rat hepatocytes high affinity taurocholic acid derivatives were synthesized. One type was designed for coupling to a Sepharose matrix and the other for copolymerization with acrylamide. The resulting matrices were found to be suitable for affinity chromatography and affinity electrophore...

In evaluating the downstream processing of genetic engineered human insulin there has been no plausible correlation between the possible yield (determined via sodium dodecyl sulphate polyacrylamide gel electrophoresis) and the actual yield, i.e., the pure product on the balance. The aim of this work was to develop an immunological method for the de...

Antibodies were raised against the carboxy terminus of the insulin A chain in sheep, goat and rabbit using as antigen the synthetic octapeptide YQLENYCN conjugated to bovine serum albumin (BSA). All of the antisera obtained cross-reacted with molecules containing this peptide sequence. Using these antibodies, we developed a radioimmunoassay that co...

Neonatal hepatocytes are less active in uptake of bile acids than are mature hepatocytes. This phenomenon has been further investigated by transport studies with azidobenzamidotaurocholate (ABATC). Taurocholate, cholate and the photolabile ABATC were taken up by liver cells of adult rats by a sodium-dependent and by an additional sodium-independent...

In the rat hepatocellular transport system proteins with a molecular weight of 50 kDa and 54 kDa have been identified as components of the bile acid uptake system (Ziegler et al., 1984; von Dippe et al., 1986; Fricker et al., 1987; Müllner et al., 1988; Ziegler et al., 1988)

Substitution of the hydroxyl group on C7 of cholic acid by a benzamido group leads to a derivative with inhibiting quality for the inward transport of both bile acids and phallotoxins by isolated liver cells. The tritiated isothiocyanate derivative was prepared (3'- isothiocyanatobenzamido [3H]cholate, [3H] IBCA ) with a specific activity of 70-80...