scholarly journals Orphan Designation and Cisplatin/Hyaluronan Complex in an Intracavitary Film for Malignant Mesothelioma

Pharmaceutics ◽  
2021 ◽  
Vol 13 (3) ◽  
pp. 362
Author(s):  
Sabrina Banella ◽  
Eride Quarta ◽  
Paolo Colombo ◽  
Fabio Sonvico ◽  
Antonella Pagnoni ◽  
...  
Keyword(s):  
Sodium Hyaluronate ◽  
Chloride Ions ◽  
Complete Resection ◽  
Size Exclusion ◽  
European Medicines Agency ◽  
Reverse Phase Hplc ◽  
Reverse Phase ◽  
Pharmaceutical Development ◽  
Film Forming ◽  
Exclusion Chromatography

Pleural mesothelioma is a lung diffuse tumor, whose complete resection is unlikely. Consequently, metastases reappear where the primary tumor was removed. This paper illustrates the orphan medicine designation procedure of an intracavitary cisplatin film and related pharmaceutical development aspects requested by the European Medicines Agency (EMA) in its Scientific Advice. Since cisplatin pharmacokinetics from the implanted film in sheep resulted substantially modified compared to intravenous administration, the formation of a cisplatin/hyaluronan complex had been hypothesized. Here, the interaction between sodium hyaluronate (NaHA) and cisplatin (CisPt) was demonstrated. Size exclusion chromatography qualitatively evidenced the complex in the film-forming mixture, only showing the NaHA peak. Atomic absorption spectroscopy of the corresponding fraction revealed platinum, confirming the interaction. Reverse phase HPLC quantified about 5% free cisplatin in the film-forming mixture, indirectly meaning that 95% was complexed. Finally, a study of CisPt release from the film assessed how CisPt/NaHA complex affected drug availability. In water, a medium without chloride ions, there was no release and the film remained intact for 48 h and longer, whereas the placebo film dissolved in 15 min. In 0.9% NaCl medium, the film became more soluble, dissolving within 3–4 h. However, cisplatin release was still controlled by the existing complex in solution until chloride ions displaced it. While the film modified its dissolution with aging, CisPt release remained unaffected (90% released in 48 h).

scholarly journals Purification of bacteriocins using size-exclusion chromatography

2016 ◽  
Vol 11 (2) ◽  
pp. 281 ◽  
Author(s):  
Vivek K. Bajpai ◽  
Irfan A. Rather ◽  
Alshammari Fanar Hamad
Keyword(s):  
Mass Spectrometry ◽  
Ammonium Sulfate ◽  
Size Exclusion Chromatography ◽  
Size Exclusion ◽  
Ammonium Sulfate Precipitation ◽  
Reverse Phase Hplc ◽  
Reverse Phase ◽  
Cell Free Supernatant ◽  
Video Clips ◽  
Exclusion Chromatography

<p>The bacteriocin purification involves following main steps. a). Extraction of cell-free-supernatant of bacteria. b). Ammonium sulfate precipitation. c). Dialysis. d). Diafiltration using PVP and e). Size-exclusion chromatography. However, depending on the nature of work, the compound could be further analyzed by reverse-phase HPLC, NMR, mass spectrometry and sequencing.</p><p><strong>Video Clips</strong></p><p><a href="https://youtube.com/v/u1BmWfOTS9w">Part 1</a>: 4 min 52 sec</p><p><a href="https://youtube.com/v/aF45JFnwErc">Part 2</a>: 1 min 47 sec</p>

scholarly journals Identification of food-derived peptides in human blood after ingestion of corn and wheat gluten hydrolysates

2018 ◽  
Vol 2 ◽  
Author(s):  
Akika Ejima ◽  
Megumi Nakamura ◽  
Yasushi A. Suzuki ◽  
Kenji Sato
Keyword(s):  
Matrix Protein ◽  
Leucine Aminopeptidase ◽  
Wheat Gluten ◽  
Protein Hydrolysates ◽  
The Body ◽  
Size Exclusion ◽  
Extracellular Matrix Protein ◽  
Reverse Phase Hplc ◽  
Before And After ◽  
Exclusion Chromatography

Bioactive peptides in the body after ingestion of plant protein hydrolysates have been speculated but not identified. We aimed to establish an approach to identify small amounts of food-derived peptides in humans after ingestion of non-extracellular matrix protein hydrolysates. Corn and wheat gluten hydrolysates were digested using pancreatin and leucine aminopeptidase; the resultant peptides were identified via size-exclusion chromatography and reverse-phase HPLC-tandem mass spectrometry (MS/MS). Structures of indigestible peptides were confirmed via LC-MS/MS in multi-reaction monitoring mode. All indigestible peptides in the exopeptidase digest were diprolyl and di- and tripyroglutamyl peptides. Blood collected from healthy volunteers (n = 4) before and after ingestion of 9 g of the hydrolysates was assessed for the indigestible peptides via LC-MS/MS. Six peptides (Pro-Ala, Pro-Gly, Pro-Gln, pyroGlu-Pro, pyroGlu-Leu-Pro, and pyroGlu-Gln-Pro) significantly increased in human plasma up to 10–100 nM compared to the baseline. This may hence be a powerful tool for identifying foodderived peptides in blood.

scholarly journals Multidimensional profiling of plasma lipoproteins by size exclusion chromatography followed by reverse-phase protein arrays

2011 ◽  
Vol 52 (12) ◽  
pp. 2323-2331 ◽  
Author(s):  
Gregor Dernick ◽  
Stefan Obermüller ◽  
Cyrill Mangold ◽  
Christine Magg ◽  
Hugues Matile ◽  
...  
Keyword(s):  
Size Exclusion Chromatography ◽  
Size Exclusion ◽  
Plasma Lipoproteins ◽  
Reverse Phase ◽  
Protein Arrays ◽  
Phase Protein ◽  
Exclusion Chromatography

scholarly journals Coupling Multi-Angle Light Scattering to Reverse-Phase Ultra-High-Pressure Chromatography (RP-UPLC-MALS) for the characterization monoclonal antibodies

2019 ◽  
Vol 9 (1) ◽  
Author(s):  
Lorenzo Gentiluomo ◽  
Vanessa Schneider ◽  
Dierk Roessner ◽  
Wolfgang Frieß
Keyword(s):  
High Pressure ◽  
Light Scattering ◽  
Physical Stability ◽  
Exchange Chromatography ◽  
Protein Characterization ◽  
Size Exclusion ◽  
Reverse Phase ◽  
Long Term Stability ◽  
Ultra High Pressure ◽  
Exclusion Chromatography

Abstract Multi-angle light scattering coupled with size-exclusion chromatography (SEC-MALS) is a standard approach for protein characterization. Recently MALS detection has been coupled with ion-exchange chromatography (IEX) which demonstrated the feasibility and high value of MALS in combination with non-sized-based fractionation methods. In this study we coupled reverse-phase ultra-high pressure liquid chromatography (RP-UPLC) with a low-dispersion MALS detector for the characterization of intact monoclonal antibody (mAbs) and their fragments. We confirmed a constant refractive index increment value for mAbs in RP gradients, in good agreement with the values in literature for other classes of proteins. We showed that the impurities eluting from a RP column can often be related to aggregated species and we confirmed that in most cases those oligomers are present also in SEC-MALS. Yet, in few cases small aggregates fractions in RP-UPLC are an artifact. In fact, proteins presenting thermal and physical stability not suitable for the harsh condition applied during the RP separation of mAbs (i.e. organic solvents at high temperature) can aggregate. Further, we applied RP-UPLC-MALS during a long term stability studies. The different principle of separation used in RP-UPLC- MALS provides an additional critical level of protein characterization compared to SEC-MALS and IEX-MALS.

scholarly journals Biochemical and Pharmacological Characterization of TLBbar, a New Serine Protease with Coagulant Activity from Bothrops barnetti Snake Venom

2013 ◽  
Vol 2013 ◽  
pp. 1-11
Author(s):  
Magaly Alejandra Brousett-Minaya ◽  
Paulo Aparecido Baldasso ◽  
Salomón Huancahuire-Vega ◽  
Sérgio Marangoni
Keyword(s):  
Serine Protease ◽  
Snake Venom ◽  
Serine Proteases ◽  
Kinetic Studies ◽  
Size Exclusion ◽  
Reverse Phase Hplc ◽  
Pharmacological Characterization ◽  
Coagulant Activity ◽  
Exclusion Chromatography

A thrombin-like enzyme named TLBbar was isolated from Bothrops barnetti snake venom and its biochemical and pharmacological characteristics were determined. TLBbar was purified using size exclusion chromatography and reverse phase HPLC, showing molecular mass of 28750.7 Da determined by mass spectrometry. TLBbar serine protease is basic (pI 7.4) and its structure shows similarity with other serine proteases of snake venom. Optimal proteolytic activity was at 37°C and pH 8; this activity was strongly inhibited by PMSF and Leupeptin, however; heparin, and soybean trypsin inhibitor (SBT-I) were ineffective. Kinetic studies on BApNA chromogenic substrate have revealed that TLBbar presents a Michaelis-Menten kinetics, with values of Km and Vmax of 0.433 mM and 0.42 nmol/min, respectively. TLBbar showed high clotting activity upon bovine and human plasma, presenting IC of 125 and minimum dose coagulant (MDC) of 2.23 μg/μL. TLBbar cleavages the Aα chain of bovine fibrinogen, with maximal efficiency at 30–40°C in the presence of calcium after two hours incubation; this fibronogenolityc activity was inhibited by PMSF and Leupeptin, confirming its classification in the group of serine proteases. In addition, TLBbar is capable of aggregating platelets in the same way that thrombin in concentrations of 2.5 μg/μL.

Size exclusion chromatography of polysaccharides with reverse phase liquid chromatography

2014 ◽  
Vol 1323 ◽  
pp. 97-103 ◽  
Author(s):  
Yan He ◽  
Weiying Hou ◽  
Melissa Thompson ◽  
Heidi Holovics ◽  
Tracy Hobson ◽  
...  
Keyword(s):  
Liquid Chromatography ◽  
Size Exclusion Chromatography ◽  
Size Exclusion ◽  
Reverse Phase ◽  
Reverse Phase Liquid Chromatography ◽  
Exclusion Chromatography ◽  
Phase Liquid
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