Using Analytical Quality by Design to improve analytical method development in vaccines quality control: Application to an optimized quantitative high-performance anion-exchange chromatographic method.

Analytical quality by design (AQbD) is an enhanced approach for the development of analytical methods. AQbD has received much industrial interest, being the subject of several recently published draft guidelines. This article demonstrates the application of AQbD to determine the quantity of non-adsorbed polysaccharide polyribosyl ribitol phosphate (PRP) and percentage of depolymerized PRP in a commercial hexavalent liquid vaccine, and establishment of an analytical control strategy (ACS). The quantification method developed is high-performance anion-exchange chromatography (HPAEC) with pulsed amperometric detection, preceded by ultracentrifugation (sample preparation) for separation of the depolymerized polysaccharide from the native adsorbed polysaccharide. The first step was to develop the analytical target profile (ATP) which defines the purpose of the analytical measurement as well as the development scope. As a second step, risk assessment tools were used for identification and ranking of the critical method variables (CMVs) which have a potential impact on method performance if not controlled. Based on a multivariate Design of Experiments (DoE) approach, a proposed method operational design region (MODR) was determined for seven CMVs. Finally, the ACS was established from the understanding of the analytical method and the robustness study. This article focuses on robust and operational ranges of critical parameters linked to the ultracentrifugation and chromatographic steps for depolymerized polysaccharide content control. The design space proposed for CMVs corresponds to the ranges that ensure a product that complies with the previously established precision criteria (±2% equivalent to ± 10 % around the product criterion, which is 20 % for depolymerized polysaccharide control limit). The following design space was established from the DoE statistical modeling for ultracentrifugation critical parameters: [483,000-520,000] g for speed, [11-19]°C for temperature, [29-34] minutes for duration, and from extemporaneous to 8 min for holding time before supernatant recuperation after the ultracentrifugation. For chromatographic critical parameters, the MODR is [2-6] psi for mobile phase helium pressure, [0-7] days for mobile phase storage time, and [0-3] days for samples storage time in the autosampler at 5 °C. Methods optimized using the AQbD approach provide strong justifications during regulatory filing for the selection of analytical CMVs, and for the ACS to be applied during the lifecycle management of the method.

Holistic analytical characterization and risk assessment of residual host cell protein impurities in an active pharmaceutical ingredient synthesized by biocatalysts.

Host cell proteins (HCPs) are a significant class of process-related impurities commonly associated with the manufacturing of biopharmaceuticals. However, due to the increased use of crude enzymes as biocatalysts for modern organic synthesis, HCPs can also be introduced as a new class of impurities in chemical drugs. In both cases, residual HCPs need to be adequately controlled to ensure product purity, quality, and patient safety. Although a lot of attentions have been focused on defining a universally acceptable limit for such impurities, the risks associated with residual HCPs on product quality, safety, and efficacy often need to be determined on a case-by-case basis taking into consideration the residual HCP profile in the product, the dose, dosage form, administration route, and so forth. Here we describe the unique challenges for residual HCP control presented by the biocatalytic synthesis of an investigational stimulator of interferon genes protein agonist, MK-1454, which is a cyclic dinucleotide synthesized using Escherichia coli cell lysate overexpressing cyclic GMP-AMP synthase as a biocatalyst. In this study, a holistic characterization of residual protein impurities using a variety of analytical tools including nanoscale liquid chromatography coupled to tandem mass spectrometry, together with in silico immunogenicity prediction of identified proteins, facilitated risk assessment and guided process development to achieve adequate removal of residual protein impurities in MK-1454 active pharmaceutical ingredient.

Proposal of a simple and rapid method for the chemotherapy preparations analytical control by spectrophotometry UV-Vis method.

INTRODUCTION: The preparation of anticancer chemotherapy in a hospital must meet several objectives; the first main is the quality, which can be provided by setting up a surveillance system and a quality control of each preparations. The aim of this work is to present a simple, fast and accurate spectrophotometric method for the routine control of cytotoxic preparations. MATERIALS AND METHODS: This is a study carried out in the cytotoxic preparation unit of the university center of Rabat-Sale children's hospital in Morocco. All samples of preparations were collected and analyzed daily on the site. After validation of the analytical method with respect to many parameters such as: linearity, accuracy and precision according to ICH Q2 guidelines, samples of cytotoxic preparations collected were assayed. RESULTS: The results are satisfactory with good level of exactitude, and high precision. CONCLUSION: Compared to other techniques, this method can be considered as a useful alternative in the routine quality control of preparations. It can quickly obtain qualitative and quantitative information with instrumentation and inexpensive reagents.

Scale-Up of Plasmid DNA Downstream Process Based on Chromatographic Monoliths.

Purification of high-quality plasmid DNA in large quantities is a crucial step in its production for therapeutic use and is usually conducted by different chromatographic techniques. Large-scale preparations require the optimization of yield and homogeneity, while maximizing removal of contaminants and preserving molecular integrity. The advantages of Convective Interaction Media® (CIM®) monolith stationary phases, including low backpressure, fast separation of macromolecules, and flow-rate-independent resolution qualified them to be used effectively in separation of plasmid DNA on laboratory as well as on large scale. A development and scale-up of plasmid DNA downstream process based on chromatographic monoliths is described and discussed below. Special emphasis is put on the introduction of process analytical technology principles and tools for optimization and control of a downstream process.

Physical and chemical analysis of commercial nystatin / Análises físico-químicas da nistatina comercial

Nystatin (NYS) is a fermentation-produced antibiotic of the polyene group. Commercial NYS is a mixture of compounds named NIS A1, A2 and A3. Current analysis undertook physical and chemical analyses in two samples of commercial nystatin (NYS I and NYS II). Moisture (Karl Fischer), spectroscopic (IR, UV and fluorescence) and thermal analyses (TGA and DSC) were conducted. The moisture was respectively 9.2% and 8.8% for samples I and II. Absorption spectrum in the UV/VIS region had a vibronic structure with three max.. The mirror image rule was not complied with in the fluorimetric analysis. Spectroscopy analysis in the IV region indicated that samples showed spectra similar one to another, analogous to crystal type A. Thermal analyses by DSC provided a wide, single endothermal peak and, therefore, similarities among the samples. DTG shows that samples undergo decomposition at three phases within the temperature range under analysis. Results show that samples are impure and not polymorphic, constituted by a mixture of the compounds A1, A2 and A3.

A nistatina (NIS) é um antibiótico do grupo dos poliênicos produzido por fermentação. A NIS comercial consiste numa mistura de compostos denominados NIS A1, A2 e A3. O presente trabalho teve por objetivo realizar análises físico-químicas de duas amostras de nistatina comercial (NIS I e NIS II). Foram realizadas análises de umidade (Karl Fischer), espectroscópicas (IV, UV e fluorescência) e térmicas (TGA e DSC). A umidade encontrada foi de 9,2% e 8,8% para as amostras I e II, respectivamente. O espectro de absorção na região do UV/VIS apresentou estrutura vibrônica com três máxs.. Na análise fluorimétrica não se observou obediência à regra da imagem espelho. A análise por espectroscopia na região do IV mostrou que as amostras apresentam espectros semelhantes entre si, análogos ao cristal do tipo A. As análises térmicas por DSC apresentaram um pico endotérmico, largo, único, sugerindo semelhanças entre as amostras. O DTG indica que as amostras sofrem decomposição em três fases no intervalo de temperatura estudado. Dos resultados obtidos, concluiu-se que que as amostras são impuras e não polimorfas, constituindo-se de uma mistura dos compostos A1, A2 e A3.