Conjugation and Other Methods in Polymeric Vaccines
Mesut Karahan in Synthetic Peptide Vaccine Models, 2021
Chromatography is known as an important biophysical technique that can separate, identify, and purify the blend segments for subjective and quantitative investigation. Proteins can be purified based on properties such as size and shape, total charge, hydrophobic groups present on the surface, and the ability to bind to the stationary phase. It is based on molecular properties and interaction type use mechanism, four separation technologies, ion exchange, dispersion, surface adsorption, and size exclusion. Column chromatography is one of the most used and common techniques for protein purification methods. This technique is basically used to purify biological molecules. The application of the method can be summarized as follows. The sample is separated on the column (stationary phase) and then the wash buffer is added to the column (mobile phase). It flows through the column material placed on the fiberglass support. With the help of the wash buffer, the samples are accumulated at the bottom of the column chromatography instrument, based on time and volume (Coskun 2016). Column chromatography is a powerful purification and separation process that is closely controlled to the hydrodynamic diameters of the macromolecules depending on the diameter of the pores in the filling material (see in HPLC Method) (Acar 2006; Fornaguera and Solans 2018).
Dictionary
Mario P. Iturralde in Dictionary and Handbook of Nuclear Medicine and Clinical Imaging, 1990
Chromatography. This method of chemical analysis uses a group of methods for separating a mixture of substances into their component parts. Chromatography is a misnomer for the technique since the color of the components is not really used for identifying substances in modern techniques. There are four basic types of chromatography using one phase or substrate which may be liquid or solid through which the test substance (gas or liquid) moves. A common feature of these methods is that the difference in rate of movement of the components of the test sample (plus reagents) is used to identify the separate components. In the clinical laboratory these methods are used to detect complex substances such as drugs and hormones. Examples of chromatographs are the gas-liquid Chromatograph (GLC) and the thin-layer Chromatograph (TLC). Both of these are being superseded by high-performance liquid chromatography (HPLC), although some compounds can only be measured by GLC.
Biologic Drug Substance and Drug Product Manufacture
Anthony J. Hickey, Sandro R.P. da Rocha in Pharmaceutical Inhalation Aerosol Technology, 2019
Passage of the clarified medium containing the target mAb through a column consisting of staphylococcal protein A (called SPA, or simply protein A), or its smaller ligands, immobilized on porous beads, leads to selective binding and isolation of the antibody. This process is called affinity chromatography, since the separation is driven by the specific affinity of the stationary phase for the desired component of the mobile phase. SPA is a cell-wall associated protein domain on the surface of S. aureus. The porous beads are typically made of a resin that is acid and base resistant to allow for efficient column handling in different pH environments such as those commonly encountered for elution (low pH) and clean-in-place protocols (high pH).
Enhanced oral bioavailability of koumine by complexation with hydroxypropyl-β-cyclodextrin: preparation, optimization, ex vivo and in vivo characterization
Published in Drug Delivery, 2021
Qing Hu, Xiaoling Fu, Yanping Su, Yanfang Wang, Sihuan Gao, Xiaoqin Wang, Ying Xu, Changxi Yu
Plasma samples were prepared by the protein precipitation method (Chen et al. 2013). Briefly, an aliquot of 50 μL plasma sample and 200 μL of acetonitrile containing 10 ng/mL gelsemine (internal standard, IS) were added to a centrifuge tube. The mixture was vortexed for 5 min and centrifuged at 18,000 rpm for 10 min at 4 °C. Then the supernatant was transferred and injected into the UPLC-MS/MS system for analysis. Chromatographic separation was performed using a UPLC instrument (Agilent 1290, Agilent Technologies, USA), equipped with a mass spectrometer (QTRAP® 5500, AB SCIEX, USA). The mass spectrometer detector was performed with multiple reactions monitoring scan mode at m/z 307.2 → 180.1 for KME and m/z 323.1 → 236.1 for IS. More detailed information about UPLC-MS/MS is provided in the supplementary material. The main pharmacokinetic parameters such as T1/2, Cmax, Tmax, and area under the curve (AUC), were determined using the non-compartmental model with DAS 3.2.1 software. The relative bioavailability F was calculated by the following equation:
Using online content uniformity measurements for rapid automated process development exemplified via an X-ray system
Published in Pharmaceutical Development and Technology, 2019
Bernhard Wagner, Thomas Brinz, Johannes Khinast
Machines for the online gravimetric weighing are commercially available from several manufactures and can detect the capsule weight within the RAPD concept. Compared to this, a CU check requires much more effort and equipment. On the one hand, there are the analytical techniques like for example HPLC (high performance liquid chromatography), TLC (thin layer chromatography), or GC (gas chromatography). These techniques require an elaborated sample preparation before the content can be determined. The measurements take a certain time and the content cannot be measured in-line within the capsule production. Thus, an integration into the RAPD would be challenging, as a requirement is a fast measurement of the samples in an automated way. Possibly, the measurements can be automated (e.g. Saitoh and Yoshimori 2008) or executed offline. However, significant time and effort is required.
A review on human body fluids for the diagnosis of viral infections: scope for rapid detection of COVID-19
Published in Expert Review of Molecular Diagnostics, 2021
Sphurti S Adigal, Nidheesh V Rayaroth, Reena V John, Keerthilatha M Pai, Sulatha Bhandari, Aswini Kumar Mohapatra, Jijo Lukose, Ajeetkumar Patil, Aseefhali Bankapur, Santhosh Chidangil
Several studies have reported the use of different spectroscopic techniques for early diagnosis of various diseases using different body fluids as clinical specimen [91]. High-performance liquid chromatography (HPLC) is considered as a versatile tool for separation and analysis of the biological and pharmaceutical compounds. Chromatography column is the heart of chromatography technique and hence a column selection is important to separate the mixture of components in a sample of interest [92]. Generally, chromatography-based techniques are used as standard techniques by industrial sectors, federal agencies, academies, and food and drug administration (FDA). Using chromatography-based-techniques the limit of detection (LOD) for melamine was achieved to the ppb level [93]. The combination of ultrasensitive optical technique Laser-Induced Fluorescence (LIF), with highly efficient separation technique such as HPLC, detection of ultra-trace quantities of individual biomolecules in complex, multicomponent physiological systems is feasible [94]. Our earlier studies demonstrated the capability of HPLC-LIF for protein profile analysis of micro-quantities of clinical samples such as saliva [95], serum [96], cellular samples [16] and tissue homogenates [97] for the diagnosis of cancers of different types.
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