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- 实验方案
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Process Monitoring of Virus-like Particle Reassembly by Diafiltration with UV/Vis Spectroscopy and Light Scattering
摘要： Virus-Like Particles (VLPs) have shown great potential as biopharmaceuticals on the market and in clinics. Non-enveloped, in vivo-assembled VLPs are typically dis- and reassembled in vitro to improve particle stability, homogeneity, and immunogenicity. At industrial scale, Cross-Flow Filtration (CFF) is the method of choice for performing reassembly by diafiltration. Here, we developed an experimental CFF setup with on-line measurement loop for the implementation of Process Analytical Technology (PAT). The measurement loop included an Ultraviolet and Visible (UV/Vis) spectrometer as well as a light-scattering photometer. These sensors allowed for monitoring protein concentration, protein tertiary structure, and protein quaternary structure. The experimental setup was tested with three Hepatitis B core Antigen (HBcAg) variants. With each variant, three reassembly processes were performed at different Transmembrane Pressures (TMPs). While light scattering provided information on the assembly progress, UV/Vis allowed for monitoring the protein concentration and the rate of VLP assembly based on the microenvironment of Tyrosine-132. VLP formation was verified by off-line Dynamic Light Scattering (DLS) and Transmission Electron Microscopy (TEM). Furthermore, the experimental results provided evidence of aggregate-related assembly inhibition and showed that off-line Size-Exclusion Chromatography (SEC) does not provide a complete picture of the particle content. Finally, a Partial-Least Squares (PLS) model was calibrated to predict VLP concentrations in the process solution. Q2 values of 0.947 to 0.984 were reached for the three HBcAg variants. In summary, the proposed experimental setup provides a powerful platform for developing and monitoring VLP reassembly steps by CFF.
关键词： process analytical technology,self-assembly,cross-flow filtration,virus-like particles,downstream processing,spectroscopy
更新于2025-09-19 17:15:36
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Application of infrared spectroscopy as Process Analytics Technology (PAT) approach in biodiesel production process utilizing Multivariate Curve Resolution Alternative Least Square (MCR-ALS)
摘要： Process Analytical Technology means at-line collection of analytical information from the process when the reaction is in progress. Obtained information enables process engineers to better control the Critical Process Parameters and direct the reaction to desirable routs. Near-infrared spectroscopy due to its analytical features, as well as the high capability of automation, versatile sampling and spectral acquisition methods is a useful tool in process monitoring when coupled to chemometrics. The Multiple Scatter Correction preprocessing technique and Alternative Least Square method can extract spectral and concentration information of a reaction mixture simultaneously, were employed pairing with fiber optic Near-Infrared spectroscopy in 1000-2250 nm spectral region was employed to extract the analytical information from the biodiesel production process. The obtained output demonstrated appropriate results about the concentration and type of components during the process. The kinetics of the reaction was also studied using the obtained results and showed a three-stage kinetics with different rates.
关键词： MCR-ALS,Near-Infrared spectroscopy,Process Analytical Technology,Biodiesel
更新于2025-09-19 17:15:36
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Prediction of differential scanning calorimetry curve of theophylline direct compression model tablet using Raman spectra
摘要： The aim of this study was to predict Differential Scanning Calorimetry (DSC) curves of pharmaceutical direct compression model tablet by chemometrics using Raman spectroscopy, measured in a nondestructive and contact-free manner. Theophylline anhydrate, theophylline hydrate, and micro-crystalline cellulose were ground and mixed with a mortar and pestle to obtain various formulations. The tablets were then measured with a handheld Raman spectrometer. DSC curves of the ground powder were measured, and the first-order derivative DSC curves were calculated. The Partial Least Squares (PLS) models were constructed to predict the calories at each temperature from the Raman spectra, and the regression vector of the constructed PLS models was plotted with 2D mapping (Raman wavenumber, Temperature, Regression coefficient). This mapping system allowed for the analysis of the correlations between Raman shift and DSC endothermic peaks.
关键词： theophylline,pseudo polymorphism,Raman spectroscopy,Partial least squares,thermal analysis,process analytical technology
更新于2025-09-10 09:29:36
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Fluorescence-based analyser as a rapid tool for determining soluble protein content in dairy ingredients and infant milk formula
摘要： Milk protein, in particular native whey protein, is of interest to dairy manufacturers as a measure of functional and nutritional quality. However, quantification of soluble whey protein (SP) is time consuming; giving rise to the need to develop rapid, accurate, and portable at-line process analytical technology. In this study, the performance of a fluorescence-based analyser(F) (Amaltheys II, Spectralys Innovations, France) was evaluated for quantification of SPF and whey protein nitrogen index (WPNI)F in skim milk, whey protein concentrate and infant formula powders. Rehydration of powders prior to analysis was a key factor for ensuring repeatability and reproducibility. A comparison of the analyser with reference methods for SPF and WPNIF resulted in coefficient of determination (R2) > 0.993 for both SPKjeldahl method and WPNIGEA. The results show the fluorescence-based analyser to be rapid, compact, and accurate device, suited for providing reliable support to dairy ingredient and infant formula manufacturers.
关键词： Infant milk formula,Process analytical technology,Dairy ingredients,Whey protein nitrogen index,Soluble protein,fluorescence-based analyser
更新于2025-09-09 09:28:46
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Raman Spectroscopy for Process Analytical Technologies of Pharmaceutical Secondary Manufacturing
摘要： As the process analytical technology (PAT) mindset is progressively introduced and adopted by the pharmaceutical companies, there is an increasing demand for effective and versatile real-time analyzers to address the quality assurance challenges of drug manufacturing. In the last decades, Raman spectroscopy has emerged as one of the most promising tools for non-destructive and fast characterization of the pharmaceutical processes. This review summarizes the achieved results of the real-time application of Raman spectroscopy in the field of the secondary manufacturing of pharmaceutical solid dosage forms, covering the most common secondary process steps of a tablet production line. In addition, the feasibility of Raman spectroscopy for real-time control is critically reviewed, and challenges and possible approaches to moving from real-time monitoring to process analytically controlled technologies (PACT) are discussed.
关键词： secondary pharmaceutical manufacturing,process analytical technology,process analytically controlled technology,Raman spectroscopy
更新于2025-09-04 15:30:14