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Affinity Matrices (affinity + matrix)

Distribution by Scientific Domains

Chemistry	80%

Selected Abstracts

Differential interactions of plasmid DNA, RNA and genomic DNA with amino acid-based affinity matrices

JOURNAL OF SEPARATION SCIENCE, JSS, Issue 17-18 2010
Angela Sousa
Abstract The development of a strategy to plasmid DNA (pDNA) purification has become necessary for the development of gene therapy and DNA vaccine production processes in recent years, since this nucleic acid and most of contaminants, such as RNA, genomic DNA and endotoxins, are negatively charged. An ideal separation methodology may be achieved with the use of affinity interactions between immobilized amino acids and nucleic acids. In this study, the binding behaviour of nucleic acids under the influence of different environmental conditions, such as the composition and ionic strength of elution buffer, and the temperature, is compared with various amino acids immobilized on chromatography resins. Supercoiled (sc) plasmid isoform was isolated with all matrices used, but in some cases preferential interactions with other nucleic acids were found. Particularly, lysine chromatography showed to be an ideal technology mainly on RNA purification using low salt concentration. On the other hand, arginine ligands have shown a greater ability to retain the sc isoform comparatively to the other nucleic acids retention, becoming this support more adequate to sc pDNA purification. The temperature variation, competitive elution and oligonucleotides affinity studies also allowed to recognize the dominant interactions inherent to biorecognition of pDNA molecule and the affinity matrices. [source]

Congruence of individual cranial bone morphology and neutral molecular affinity patterns in modern humans

AMERICAN JOURNAL OF PHYSICAL ANTHROPOLOGY, Issue 2 2009
Noreen von Cramon-Taubadel
Abstract Recent studies have demonstrated that the shape of the human temporal bone is particularly strongly correlated with neutral genetic expectation, when compared against other cranial regions, such as the vault, face, and basicranium. In turn, this has led to suggestions that the temporal bone is particularly reliable in analyses of primate phylogeny and human population history. While several reasons have been suggested to explain the temporal bone's strong fit with neutral expectation, the temporal bone has never systematically been compared against other individual cranial bones defined using the same biological criteria. Therefore, it is currently unknown whether the shapes of all cranial bones possess reliable information regarding neutral genetic evolution, or whether the temporal bone is unique in this respect. This study tests the hypothesis that the human temporal bone is more congruent with neutral expectation than six other individual cranial bones by correlating population affinity matrices generated using neutral genetic and 3D craniometric data. The results demonstrate that while the temporal bone shows the absolute strongest correlation with neutral genetic data compared with all other bones, it is not statistically differentiated from the sphenoid, frontal, and parietal bones in this regard. Potential reasons for the temporal bone's consistently strong fit with neutral expectation, such as its overall anatomical complexity and/or its contribution to the architecture of the basicranium, are examined. The results suggest that future phylogenetic and taxonomic studies would benefit from considering the shape of the entire cranium minus those regions that deviate most from neutrality. Am J Phys Anthropol, 2009. © 2009 Wiley-Liss, Inc. [source]

Single-step affinity purification of recombinant proteins using a self-excising module from Neisseria meningitidis FrpC

PROTEIN SCIENCE, Issue 10 2008
Lenka Sadilkova
Abstract Purification of recombinant proteins is often a challenging process involving several chromatographic steps that must be optimized for each target protein. Here, we developed a self-excising module allowing single-step affinity chromatography purification of untagged recombinant proteins. It consists of a 250-residue-long self-processing module of the Neisseria meningitidis FrpC protein with a C-terminal affinity tag. The N terminus of the module is fused to the C terminus of a target protein of interest. Upon binding of the fusion protein to an affinity matrix from cell lysate and washing out contaminating proteins, site-specific cleavage of the Asp,Pro bond linking the target protein to the self-excising module is induced by calcium ions. This results in the release of the target protein with only a single aspartic acid residue added at the C terminus, while the self-excising affinity module remains trapped on the affinity matrix. The system was successfully tested with several target proteins, including glutathione-S-transferase, maltose-binding protein, ,-galactosidase, chloramphenicol acetyltransferase, and adenylate cyclase, and two different affinity tags, chitin-binding domain or poly-His. Moreover, it was demonstrated that it can be applied as an alternative to two currently existing systems, based on the self-splicing intein of Saccharomyces cerevisiae and sortase A of Staphylococcus aureus. [source]

A novel method of affinity-purifying proteins using a bis-arsenical fluorescein

PROTEIN SCIENCE, Issue 2 2000
Kurt S. Thorn
Abstract Genetically-encoded affinity tags constitute an important strategy for purifying proteins. Here, we have designed a novel affinity matrix based on the bis-arsenical fluorescein dye FlAsH, which specifically recognizes short ,-helical peptides containing the sequence CCXXCC (Griffin BA, Adams SR, Tsien RY, 1998, Science 281:269,212). We find that kinesin tagged with this cysteine-containing helix binds specifically to FlAsH resin and can be eluted in a fully active form. This affinity tag has several advantages over polyhistidine, the only small affinity tag in common use. The protein obtained with this single chromatographic step from crude Escherichia coli lysates is purer than that obtained with nickel affinity chromatography of 6xHis tagged kinesin. Moreover, unlike nickel affinity chromatography, which requires high concentrations of imidazole or pH changes for elution, protein bound to the FlAsH column can be completely eluted by dithiothreitol. Because of these mild elution conditions, FlAsH affinity chromatography is ideal for recovering fully active protein and for the purification of intact protein complexes. [source]