scholarly journals Ultrafiltration Method for Plasma Protein Binding Studies and Its Limitations

Processes ◽  
2021 ◽  
Vol 9 (2) ◽  
pp. 382
Author(s):  
Camelia-Maria Toma ◽  
Silvia Imre ◽  
Camil-Eugen Vari ◽  
Daniela-Lucia Muntean ◽  
Amelia Tero-Vescan
Keyword(s):  
Protein Binding ◽  
Plasma Protein Binding ◽  
Plasma Protein ◽  
Specific Binding ◽  
Critical Role ◽  
Volume Ratio ◽  
Binding Studies ◽  
Experimental Conditions ◽  
Key Aspects

Plasma protein binding plays a critical role in drug therapy, being a key part in the characterization of any compound. Among other methods, this process is largely studied by ultrafiltration based on its advantages. However, the method also has some limitations that could negatively influence the experimental results. The aim of this study was to underline key aspects regarding the limitations of the ultrafiltration method, and the potential ways to overcome them. The main limitations are given by the non-specific binding of the substances, the effect of the volume ratio obtained, and the need of a rigorous control of the experimental conditions, especially pH and temperature. This review presents a variety of methods that can hypothetically reduce the limitations, and concludes that ultrafiltration remains a reliable method for the study of protein binding. However, the methodology of the study should be carefully chosen.

scholarly journals Plasma protein binding in uremia: Extraction and characterization of an inhibitor

1980 ◽  
Vol 18 (1) ◽  
pp. 86-94 ◽  
Author(s):  
Thomas A. Depner ◽  
Paul F. Gulyassy ◽  
Linda A. Stanfel ◽  
Elizabeth A. Jarrard
Keyword(s):  
Protein Binding ◽  
Plasma Protein Binding ◽  
Plasma Protein

Evaluation of electrophoretic mobility shift assay as a method to determine plasma protein binding of siRNA

Bioanalysis ◽  
2019 ◽  
Vol 11 (21) ◽  
pp. 1927-1939 ◽  
Author(s):  
Carrie Rocca ◽  
Sean Dennin ◽  
Yongli Gu ◽  
Joohwan Kim ◽  
Samantha Chigas ◽  
...  
Keyword(s):  
Protein Binding ◽  
Electrophoretic Mobility Shift Assay ◽  
Electrophoretic Mobility ◽  
Plasma Protein Binding ◽  
Plasma Protein ◽  
Plasma Proteins ◽  
Small Interfering Rnas ◽  
Mobility Shift ◽  
Mobility Shift Assay

Aim: The electrophoretic mobility shift assay (EMSA) was evaluated as an alternative to ultrafiltration (UF) to assess plasma protein binding (PPB) of small interfering RNAs (siRNA) and antisense oligonucleotides (ASO). Results & methodology: EMSA analysis showed that PPB depended on siRNA and plasma concentration. Conversely, when analyzed by ultrafiltration, siRNA bound the filtration device nonspecifically and PPB remained >98% across physiologically relevant siRNA concentrations. Using EMSA, siRNA exhibited charge-based interactions with plasma proteins, while ASO remained highly bound to plasma proteins or albumin in the presence of 500 mM salt. Conclusion: PPB characteristics of siRNA and ASO can be distinguished using EMSA. Characterization of siRNA PPB by EMSA enhances our knowledge of siRNA absorption, distribution, metabolism and excretion and advanced development of RNA interference therapeutics.

Synthesis, Radiolabeling, and Characterization of Plasma Protein-Binding Ligands: Potential Tools for Modulation of the Pharmacokinetic Properties of (Radio)Pharmaceuticals

2017 ◽  
Vol 28 (9) ◽  
pp. 2372-2383 ◽  
Author(s):  
Cristina Müller ◽  
Renáta Farkas ◽  
Francesca Borgna ◽  
Raffaella M. Schmid ◽  
Martina Benešová ◽  
...  
Keyword(s):  
Protein Binding ◽  
Plasma Protein Binding ◽  
Plasma Protein ◽  
Pharmacokinetic Properties

scholarly journals Pharmacokinetics, Tissue Distribution, Plasma Protein Binding Studies of 10-Dehydroxyl-12-Demethoxy-Conophylline, a Novel Anti-Tumor Candidate, in Rats

Molecules ◽  
2019 ◽  
Vol 24 (2) ◽  
pp. 283
Author(s):  
Chengjun Jiang ◽  
Jie Li ◽  
Xianghai Cai ◽  
Nini Li ◽  
Yan Guo ◽  
...  
Keyword(s):  
Protein Binding ◽  
Tissue Distribution ◽  
Plasma Protein Binding ◽  
Plasma Protein ◽  
Equilibrium Dialysis ◽  
Free Fraction ◽  
Ultra Performance Liquid Chromatography ◽  
Sprague Dawley ◽  
Binding Studies ◽  
High Protein Binding

10-Dehydroxyl-12-demethoxy-conophylline is a natural anticancer candidate. The motivation of this study was to explore the pharmacokinetic profiles, tissue distribution, and plasma protein binding of 10-dehydroxyl-12-demethoxy-conophylline in Sprague Dawley rats. A rapid, sensitive, and specific ultra-performance liquid chromatography (UPLC) system with a fluorescence (FLR) detection method was developed for the determination of 10-dehydroxyl-12-demethoxy-conophylline in different rat biological samples. After intravenous (i.v.) dosing of 10-dehydroxyl-12-demethoxy-conophylline at different levels (4, 8, and 12 mg/kg), the half-life t1/2α of intravenous administration was about 7 min and the t1/2β was about 68 min. The AUC0→∞ increased in a dose-proportional manner from 68.478 μg/L·min for 4 mg/kg to 305.616 mg/L·min for 12 mg/kg. After intragastrical (i.g.) dosing of 20 mg/kg, plasma levels of 10-dehydroxyl-12-demethoxy-conophylline peaked at about 90 min. 10-dehydroxyl-12-demethoxy-conophyllinea absolute oral bioavailability was only 15.79%. The pharmacokinetics process of the drug was fit to a two-room model. Following a single i.v. dose (8 mg/kg), 10-dehydroxyl-12-demethoxy-conophylline was detected in all examined tissues with the highest in kidney, liver, and lung. Equilibrium dialysis was used to evaluate plasma protein binding of 10-dehydroxyl-12-demethoxy-conophylline at three concentrations (1.00, 2.50, and 5.00 µg/mL). Results indicated a very high protein binding degree (over 80%), reducing substantially the free fraction of the compound.

scholarly journals Characterization of Binding of Raltegravir to Plasma Proteins

2013 ◽  
Vol 57 (10) ◽  
pp. 5147-5150 ◽  
Author(s):  
Caroline Barau ◽  
Valérie Furlan ◽  
Yazdan Yazdanpanah ◽  
Catherine Fagard ◽  
Jean-Michel Molina ◽  
...  
Keyword(s):  
Protein Binding ◽  
Plasma Protein Binding ◽  
Plasma Protein ◽  
Plasma Proteins ◽  
Association Constant ◽  
Graphical Method ◽  
Acid Glycoprotein ◽  
Ph Increase

ABSTRACTThe objective of this study was to characterize raltegravir (RAL) binding to albumin and alpha-1-acid glycoprotein (AAG). Unbound and bound RAL were separated by ultrafiltration. The association constant (Ka) was estimated by a graphical method. In HIV-infected patients, the average plasma protein binding is 76%. RAL did not bind to AAG but bound to nonsaturable, low-affinity albumin sites with ann(number of sites) ·Kaproduct of 9.8 × 102liters/mol. A pH increase of 0.2 U led to a 2% increase in the bound fraction.

scholarly journals Characterization of plasma protein binding dissociation with online SPE-HPLC

2015 ◽  
Vol 5 (1) ◽  
Author(s):  
Ping Li ◽  
Yiran Fan ◽  
Yunlong Wang ◽  
Yaxin Lu ◽  
Zheng Yin
Keyword(s):  
Protein Binding ◽  
Plasma Protein Binding ◽  
Plasma Protein ◽  
Online Spe

scholarly journals In Silico Plasma Protein Binding Studies of Selected Group of Drugs Using TLC and HPLC Retention Data

2021 ◽  
Vol 14 (3) ◽  
pp. 202
Author(s):  
Karolina Wanat ◽  
Grażyna Żydek ◽  
Adam Hekner ◽  
Elżbieta Brzezińska
Keyword(s):  
Protein Binding ◽  
Plasma Protein Binding ◽  
Plasma Protein ◽  
Reversed Phase ◽  
Coefficient Of Determination ◽  
Retention Data ◽  
Linear Regression Models ◽  
Binding Studies ◽  
Tlc Plates ◽  
Living Organisms

Plasma protein binding is an important determinant of the pharmacokinetic properties of chemical compounds in living organisms. The aim of the present study was to determine the index of protein binding affinity based on chromatographic experiments. The question is which chromatographic environment will best mimic the drug–protein binding conditions. Retention data from normal phase thin-layer liquid chromatography (NP TLC), reversed phase (RP) TLC and HPLC chromatography experiments with 129 active pharmaceutical ingredients (APIs) were collected. The stationary phase of the TLC plates was modified with protein and the HPLC column was filled with immobilized human serum albumin. In both chromatographic methods, the mobile phase was based on a buffer with a pH of 7.4 to mimic physiological conditions. Chemometric analyses were performed to compare multiple linear regression models (MLRs) with retention data, using protein binding values as the dependent variable. In the course of the analysis, APIs were divided into acidic, basic and neutral groups, and separate models were created for each group. The MLR models had a coefficient of determination between 0.73 and 0.91, with the highest values from NP TLC data.

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