Springer protocols feed by Pharmacology/Toxicology 
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This is an RSS file. You can use it to subscribe to this data in your favourite RSS reader or to display this data on your own website or blog.Genome-Wide Gene Expression Profiling, Genotyping, and Copy Number Analyses of Acute Myeloid Leukemia Using Affymetrix GeneChips
With novel genome-wide technologies it is nowadays possible to perform detailed molecular analyses of normal and malignant tissues. Acute myeloid leukemia (AML) is a heterogeneous group of diseases with variable response to therapy. Gene expression profiling and genome-wide genotyping have recently been successfully applied to unravel the heterogeneity of AML. This chapter gives instructions and recommendations for genome-wide gene expression analyses, genotyping, and copy number analyses, as performed for AML using Affymetrix GeneChips. (Source: Springer protocols feed by Pharmacology/Toxicology)
Source: Springer protocols feed by Pharmacology/Toxicology - July 8, 2013 Category: Drugs & Pharmacology Source Type: news
Pharmacogenomics: Historical Perspective and Current Status
Pharmacogenomics and its predecessor pharmacogenetics study the contribution of genetic factors to the interindividual variability in drug efficacy and safety. One of the major goals of pharmacogenomics is to tailor drugs to individuals based on their genetic makeup and molecular profile. From early findings in the 1950s uncovering inherited deficiencies in drug metabolism that explained drug-related adverse events, to nowadays genome-wide approaches assessing genetic variation in multiple genes, pharmacogenomics has come a long way. The evolution of pharmacogenomics has paralleled the evolution of genotyping technologies,...
Source: Springer protocols feed by Pharmacology/Toxicology - July 8, 2013 Category: Drugs & Pharmacology Source Type: news
Hydrophobic Interaction Chromatography to Analyze Glycoproteins
Hydrophobic interaction chromatography (HIC) is one of many separation techniques that can be used to analyze proteins. The separation mechanism is based on the adsorption of the hydrophobic region of the protein to the hydrophobic ligands attached to the column in the presence of high salt. The proteins are then eluted by descending salt concentration. Here we describe the use of this HIC technique to evaluate the hydrophobicity of different monoclonal antibodies (mAbs) and to separate different heterogeneities that occur in mAb. (Source: Springer protocols feed by Pharmacology/Toxicology)
Source: Springer protocols feed by Pharmacology/Toxicology - April 13, 2013 Category: Drugs & Pharmacology Source Type: news
Characterization of Glycoprotein Biopharmaceutical Products by Caliper LC9 CE-SDS Gel Technology
Over the last decade, science has greatly improved in the area of protein sizing and characterization. Efficient high-throughput methods are now available to substitute for the traditional labor-intensive SDS-PAGE methods, which alternatively take days to analyze a very limited number of samples. Currently, PerkinElmer® (Caliper) has designed an automated chip-based fluorescence detection method capable of analyzing proteins in minutes with sensitivity similar to standard SDS-PAGE. Here, we describe the use and implementation of this technology to characterize and screen a large number of formulations of target glycopr...
Source: Springer protocols feed by Pharmacology/Toxicology - April 13, 2013 Category: Drugs & Pharmacology Source Type: news
Application of Capillary Electrophoresis in Glycoprotein Analysis
Capillary electrophoresis (CE) is a versatile analytical method used to characterize glycoproteins. We have used several modes of CE separation such as CE-SDS gel, imaged capillary isoelectric focusing (icIEF), and capillary zone electrophoresis (CZE) to study therapeutic glycoprotein products. CE-SDS gel is applied to characterize the glycan occupancy and number of glycosylation sites, and icIEF is used to study the charge heterogeneities due to sialic acids in glycoproteins. To further characterize the glycoprotein, removal of N-linked glycans is necessary and a CZE technique is employed to analyze each glycan moiety. Ex...
Source: Springer protocols feed by Pharmacology/Toxicology - April 13, 2013 Category: Drugs & Pharmacology Source Type: news
Assessing Fc Glycan Heterogeneity of Therapeutic Recombinant Monoclonal Antibodies Using NP-HPLC
Recombinant monoclonal antibodies (rMAbs) are becoming major human therapeutics to treat life-threatening diseases such as cancer. These rMAbs are produced using either in vitro cell culture processes or transgenic technology in animals or plants. Glycans present in the Fc region can affect functions of rMAbs. These Fc glycans are heterogeneous and impact binding of rMAbs to Fc gamma receptors (FcγRs) and C1q protein. As a result Fc glycans affect antibody-dependent cellular cytotoxicity and complement-dependent cytotoxicity of rMAbs. Thus understanding the glycan heterogeneity is necessary during the development of ...
Source: Springer protocols feed by Pharmacology/Toxicology - April 13, 2013 Category: Drugs & Pharmacology Source Type: news
Engineering the Baculovirus Genome to Produce Galactosylated Antibodies in Lepidopteran Cells
Nowadays, recombinant proteins are used with great success for the treatment of a variety of medical conditions, such as cancer, autoimmune, and infectious diseases. Several expression systems have been developed to produce human proteins, but one of their most critical limitations is the addition of truncated or nonhuman glycans to the recombinant molecules. The presence of such glycans can be deleterious as they may alter the protein physicochemical properties (e.g., solubility, aggregation), its half-life, and its immunogenicity due to the unmasking of epitopes. (Source: Springer protocols feed by Pharmacology/Toxicology)
Source: Springer protocols feed by Pharmacology/Toxicology - April 13, 2013 Category: Drugs & Pharmacology Source Type: news
N-Glycosylation Humanization for Production of Therapeutic Recombinant Glycoproteins in Saccharomyces cerevisiae
The production of therapeutic recombinant glycoproteins deals with three main issues: cost, production capacities, and glycosylation. Nowadays, such proteins are expressed in various complex expression systems (CHO, bacteria, etc.); the processes related to those production hosts are time consuming and expensive, or the question of posttranslational modifications (as glycosylation) control is still unresolved. There is a need to find an alternative approach, while maintaining high quality level: the new system must be able to add complex N-glycan structures to proteins of interest. Developed in several strains of Saccharom...
Source: Springer protocols feed by Pharmacology/Toxicology - April 13, 2013 Category: Drugs & Pharmacology Source Type: news
Glycoengineered Pichia-Based Expression of Monoclonal Antibodies
Currently, mammalian cells are the most commonly used hosts for the production of therapeutic monoclonal antibodies (mAbs). These hosts not only secrete mAbs with properly assembled two heavy and two light chains but also deliver mAbs with a glycosylation profile that is compatible with administration into humans. GlycoFi, a wholly owned subsidiary of Merck & Co., Inc., humanized the Pichia glycosylation pathway which allows it to express glycoproteins with a human-like glycan profile. This offers an alternative mAb production platform similar to mammalian hosts and in some cases it even provides more homogenous produc...
Source: Springer protocols feed by Pharmacology/Toxicology - April 13, 2013 Category: Drugs & Pharmacology Source Type: news
Engineering a Human-Like Glycosylation to Produce Therapeutic Glycoproteins Based on 6-Linked Sialylation in CHO Cells
When recombinant glycoproteins for therapeutic use are to be produced on an industrial scale, there is a crucial need for technologies that can engineer fast-growing stable cells secreting the protein drug at a high rate and with a defined and safe glycosylation profile. Current cell lines approved for drug production are essentially from rodent origin. Their glycosylation machinery often adds undesired carbohydrate determinants which may alter protein folding, induce immunogenicity, and reduce circulatory life span of the drug. Notably, sialic acid as N-acetylneuraminic acid is not efficiently added in most mammalian cell...
Source: Springer protocols feed by Pharmacology/Toxicology - April 13, 2013 Category: Drugs & Pharmacology Source Type: news
Engineering of Therapeutic and Diagnostic O-Glycans on Recombinant Mucin-Type Immunoglobulin Fusion Proteins Expressed in CHO Cells
Metabolic engineering of mammalian cells for optimized glycosylation is usually done to improve activity and the pharmacokinetic features of glycoprotein therapeutics. The field is mainly focused around engineering of N-glycans. We have created a platform in which recombinant mucin-type immunoglobulin fusion proteins are used as scaffolds for multivalent expression of O-glycans with diagnostic or therapeutic potential. The methods used to make stable CHO cell lines secreting a mucin-type fusion protein with blood group A or B determinants following expression of up to five different cDNAs are described. (Source: Springer p...
Source: Springer protocols feed by Pharmacology/Toxicology - April 13, 2013 Category: Drugs & Pharmacology Source Type: news
In Silico Models for B-Cell Epitope Recognition and Signaling
Tremendous technological advances in peptide synthesis and modification in recent years have resolved the major limitations of peptide-based vaccines. B-cell epitopes are major components of these vaccines (besides having other biological applications). Researchers have been developing in silico or computational models for the prediction of both linear and conformational B-cell epitopes, enabling immunologists and clinicians to identify the most promising epitopes for characterization in the laboratory. Attempts are also ongoing in systems biology to delineate the signaling networks in immune cells. Here we present all pos...
Source: Springer protocols feed by Pharmacology/Toxicology - April 13, 2013 Category: Drugs & Pharmacology Source Type: news
Databases and In Silico Tools for Vaccine Design
In vaccine design, databases and in silico tools play different but complementary roles. Databases collect experimentally verified vaccines and vaccine components, and in silico tools provide computational methods to predict and design new vaccines and vaccine components. Vaccine-related databases include databases of vaccines and vaccine components. In the USA, the Food and Drug Administration (FDA) maintains a database of licensed human vaccines, and the US Department of Agriculture keeps a database of licensed animal vaccines. Databases of vaccine clinical trials and vaccines in research also exist. The important vaccin...
Source: Springer protocols feed by Pharmacology/Toxicology - April 13, 2013 Category: Drugs & Pharmacology Source Type: news
Molecular Dynamics Simulations in Drug Design
This minireview focuses on recent developments in the application of molecular dynamics to drug design. Recent applications of endpoint free-energy computational methods such as molecular mechanics Poisson-Boltzmann surface area (MM-PBSA) and generalized Born surface area (MM-GBSA) and linear response methods are described. Recent progress in steered molecular dynamics applied to drug design is reviewed. (Source: Springer protocols feed by Pharmacology/Toxicology)
Source: Springer protocols feed by Pharmacology/Toxicology - April 13, 2013 Category: Drugs & Pharmacology Source Type: news
On Exploring Structure–Activity Relationships
Understanding structure–activity relationships (SARs) for a given set of molecules allows one to rationally explore chemical space and develop a chemical series optimizing multiple physicochemical and biological properties simultaneously, for instance, improving potency, reducing toxicity, and ensuring sufficient bioavailability. In silico methods allow rapid and efficient characterization of SARs and facilitate building a variety of models to capture and encode one or more SARs, which can then be used to predict activities for new molecules. By coupling these methods with in silico modifications of structures, one c...
Source: Springer protocols feed by Pharmacology/Toxicology - April 13, 2013 Category: Drugs & Pharmacology Source Type: news
