Development of UV–Vis Imaging Compatible Chromatographic Matrix with Application for Injectable Formulation Characterization
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Transport within human tissue matrices, e.g., the subcutaneous tissue, exhibits some resemblance to chromatographic processes. Here, a porous matrix comprising agarose beads compatible with UV–vis imaging was developed for a parallel piped rectangular flow cell (4 mm light path). Introduction of high-molecular weight dextrans (Mr ∼ 200000 and ∼500000) at 10% (w/v) rendered imaging possible by providing optical clearing of the turbid porous matrix, resulting in improved transmittance as well as resolution (from 400 to 180 μm) at 280 nm, as well as 520 nm. The interplay between diffusive and convective transport at 0 < Pe ≤ 28 was visualized at 280 nm upon injection of dexamethasone suspensions. Real-time UV–vis imaging showed in-flow cell the effect of incorporating ion-exchange resins on the retention of infliximab, lysozyme, and α-lactalbumin. The ion-exchange matrix may serve as a surrogate for polyelectrolytes in the subcutaneous tissue, assessing the potential role of electrostatic interactions of biotherapeutics upon injection. UV–vis imaging of size-exclusion chromatographic matrixes may be of interest in its own right and potentially develop into a characterization tool for injectables.Keywords:
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Abstract Inhibition of macrophage migration from a glass capillary is the most widely accepted and best characterized in vitro correlate of delayed hypersensitivity. Significant technical limitations of this method have been eliminated by a modified migration inhibitory factor (MIF) method in which macrophages are stabilized in an agarose droplet instead of a capillary tube. This agarose method is technically simple, permitting the rapid assay of large numbers of samples and requiring relatively small volumes of culture supernatant. Experiments comparing the capillary and agarose methods have established the validity and specificity of the agarose method for assay of MIF.
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After digestion of DNA with a restriction enzyme (Chapter 50), it is usually necessary, for both preparative and analytical purposes, to separate and visualize the products. In most cases, where the products are between 200 and 20,000 bp long, this is achieved by agarose gel electrophoresis. Agarose is a linear polymer that is extracted from seaweed and sold as a white powder. The powder is melted in buffer and allowed to cool, whereby the agarose forms a gel by hydrogen bonding. The hardened matrix contains pores, the size of which depends on the concentration of agarose. The concentration of agarose is referred to as a percentage of agarose to volume of buffer (w/v), and agarose gels are normally in the range of 0.3 to 3%. Many different apparatus arrangements have been devised to run agarose gels; for example, they can be run horizontally or vertically, and the current can be conducted by wicks or the buffer solution. However, today, the "submarine" gel system is almost universally used. In this method, the agarose gel is formed on a supporting plate, and then the plate is submerged into a tank containing a suitable electrophoresis buffer. Wells are preformed in the agarose gel with the aid of a "comb" that is inserted into the cooling agarose before the agarose has gelled. Into these wells are loaded the sample to be analyzed, which has been mixed with a dense solution (a loading buffer) to ensure that the sample sinks into the wells.
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Abstract This article provides a general introduction of materials characterization and describes the principles and applications of a limited number of techniques that are most commonly used to characterize the composition and structure of metals used in engineering systems. It briefly describes the classification of materials characterization methods including, bulk elemental characterization, bulk structural characterization, microstructural characterization, and surface characterization. Further, the article reviews the selection of materials characterization methods most commonly used with metals.
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Agarose [SeaKem HGT(P)] gels of 0.5% to 1.2% can be dried at room temperature and rehydrated to 84-85% of their original water content. Mobilities, using the 84-85% rehydrated gels, are indistinguishable from those on the original gels. This suggests that the fiber structure and effective pore size of agarose are independent on the 16% loosely bound (and probably synersed) water.
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After digestion of DNA with a restriction enzyme (Chapter 50), it is usually necessary, for both preparative and analytical purposes, to separate and visualize the products. In most cases, where the products are between 200 and 20,000 bp long, this is achieved by agarose gel electrophoresis. Agarose is a linear polymer that is extracted from seaweed and sold as a white powder. The powder is melted in buffer and allowed to cool, whereby the agarose forms a gel by hydrogen bonding. The hardened matrix contains pores, the size of which depends on the concentration of agarose. The concentration of agarose is referred to as a percentage of agarose to volume of buffer (w/v), and agarose gels are normally in the range of 0.3 to 3%. Many different apparatus arrangements have been devised to run agarose gels; for example, they can be run horizontally or vertically, and the current can be conducted by wicks or the buffer solution. However, today, the "submarine" gel system is almost universally used. In this method, the agarose gel is formed on a supporting plate, and then the plate is submerged into a tank containing a suitable electrophoresis buffer. Wells are preformed in the agarose gel with the aid of a "comb" that is inserted into the cooling agarose before the agarose has gelled. Into these wells are loaded the sample to be analyzed, which has been mixed with a dense solution (a loading buffer) to ensure that the sample sinks into the wells.
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In the application of agarose, especially to the research of modern biologic al technology, low gelling point agarose is necessary. Using water fraction of G elidium amansii as raw material, investingations into the preparation of low gel ling point agarose by methylation reaction was carried out. A modified agarose c ontaining alkylated agarose to the standard of low gelling point agarose. Its ma in properties of gel strength(264 g/cm2 1.0% gel), gelling point (30.1 ℃ 1. 5%gel) and melting point (64.8 ℃ 1.5%gel) are all similiar to those produced by Sigma company.
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