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10 Current white paper about the topic biochemistry

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Promotion of Aggregation as a Means of Assessing the Stability of Antibody Molecules

Thioflavin T aggregation assay to determine antibody stability using the FLUOstar Omega

03-11-2011

The efficiency of antibody molecules is dependent on their stability. We introduce an assay setup where stability of antibodies is measured. Shaking stress is initiated by the FLUOstar Omega microplate reader and aggregation propensity of antibody candidates is measured in 96-well format...

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Characterizing Biotherapeutics with HPLC

Smart Solutions for large Molecules

24-11-2010

As demonstrated here, there are a broad range of HPLC applications used in development, production, validation and release of protein therapeutics for human use. Some of the applications mentioned above are mandatory when submitting a new biopharmaceutical to approval at the regulatory agencies. The introduction of the first so-called biosimilars in Europe further increased the demand for highly efficient analysis methods. The continuous development of new stationary phases for the common modes of biochromatography, such as size exclusion, ion exchange and hydrophilic interaction chromatography support biochemists in increasing lab productivity and establishing high-throughput HPLC analysis not only for small molecules but also for large biopolymers...

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Increased productivity for downstream processing of biotherapeutics

16-11-2010

Polymer-based chromatographic resins featuring high mechanical and chemical stability and high protein binding capacities can help to increase the throughput and robustness of biopharmaceutical manufacturing processes. Advances in genetic engineering and cell culture technology have raised upstream productivity in the production of recombinant proteins. Increased titres of up to 10 g/l stress the need for highly efficient and robust downstream processes (DSP). Most industrial bioprocess development groups apply generic DSP platforms that are designed to purify various candidates of the same class of proteins applying the same process design. Besides various filtration and virus inactivation steps, they usually involve two or three chromatographic unit operations with orthogonal separation modes. The use of high capacity chromatographic resins can help to overcome the DSP bottleneck by increasing process throughput and robustness...

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Membrane Fluidity Measurements Using UV Fluorescence Polarization and the POLARstar Omega

23-06-2010

The POLARstar Omega microplate reader from BMG LABTECH was used to study membrane fluidity via fluorescence polarization in 96-well format. The method allows for the monitoring of large scale alterations in membrane fluidity (such as phase transitions), as well as more subtle changes in lipid dynamic. The change in polarization caused by dibucaine and propranolol were investigated in both liposomes and mitochondrial membranes...

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Real-Time Analysis of LNCaP Cell Growth in Different Media

11-01-2010

Our results suggest that the influence of androgen receptor-stimulating or -blocking substances strongly depends upon the composition of the growth medium. The use of hormone-depleted FBS in combination with phenol red-free medium is strongly recommended for monitoring stimulator effects, whereas inhibitory effects may be more readily analyzed in medium containing normal FBS. Because the mutated androgen receptor in LNCaP can also be stimulated by estrogens, progestogens, and several antiandrogens, we suggest that the stimulatory effect of normal FBS compared with hormone-reduced FBS may be explained by differences in estradiol levels (192 pg/ml in normal FBS compared with <12 pg/ml in hormone-reduced FBS) rather than by differences in progesterone levels (0.29 ng/ml in normal FBS compared with 0.13 ng/ml in hormone-reduced FBS) or testosterone levels (0.08 ng/ml in normal FBS compared with 0.03 ng/ml in hormone-reduced FBS). Our results show that the optimal time point for monitoring inhibitory and stimulat...

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Increased Vitality of Primary Mouse Hepatocytes Isolated with Liberase TM Research Grade

07-12-2009

As drugs applied to organisms are metabolized via the liver, hepatocytes represent a useful and common study model. Primary hepatocytes are widely used to evaluate the toxic effects of drugs and xenobiotics. A variety of metabolic parameters have been described to analyze the functionality of hepatocytes. Via those parameters, the toxic effect of substances can easily be quantified. One drawback of primary hepatocyte culture is the short duration of survival and the high sensibility of the cells. Therefore, every step towards a more gentle method of isolating the cells increases cell number and vitality and thus improves the outcome of biomedical research applications...

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HelpCorner - Transfection

31-08-2009

Transfection is the delivery of DNA, RNA, proteins, and macromelecules into eukaryotic cells. The goals for transfection include the study of gene function and regulation as well as protein expression and function. It is also used for gene silencing and protein production. According to the application, the information can be either transiently (transient transfection) or permanently expressed (stable transfection). Based on the strategies, the transfection methods can be divided into three groups:...

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Isolation of Keratinocytes and Fibroblasts from Human Foreskin by One-step Enzyme Incubation Using Liberase Research Grade Products

05-06-2009

For the in vitro testing of pharmaceutical, cosmetic and chemical products conventional cell cultures are increasingly being replaced by three-dimensional (3D) cell culture models with organ-specific characteristics. Using tissue engineering techniques, a living human skin equivalent has been developed at the Fraunhofer IGB which mimics the normal human skin. The fabrication of the 3D skin model requires the isolation and growth of primary keratinocytes and fibroblasts from human skin tissue. For the isolation of the cells it is necessary to dissociate the skin tissue in the dermal and epidermal layer without destroying the cells. Usually, cell isolation is performed in a two-step protocol. Following dissociation of both layers with dispase, collagenases are used to release the fibroblasts from the dermis. The goal of all cell isolation procedures is to maximize the yield of dissociated cells that are viable and functionally active....

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Bradford Assay Performed on BMG LABTECH´s FLUOstar Omega with new Evaluation Software

28-02-2008

Determining the protein concentration of samples is a necessary and often used method in biochemistry. Different colorimetric protein assays have been developed. The most commonly used methods are the Bradford assay, the Lowry assay and the BCA assay. In this application note we demonstrate how to determine the protein concentration of samples by using the Bradford assay and the new FLUOstar Omega....

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From Sephadex to GE Healthcare

29-09-2004

The success of chromatography as a technique for separating biomolecules is inevitably linked to the introduction of the gel filtration medium Sephadex(TM) by Pharmacia AB in 1959...

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