A Sequential Ultrafiltration Method to Enhance the Accuracy and Throughput in Plasma Protein Binding Tests

<b>Objectives</b>: Ultrafiltration (UF) is widely accepted as a method for assessing the plasma protein binding (PPB) of drugs. However, it is vulnerable to non-specific binding (NSB) to the device, which can result in inaccuracies. This study presents a straightforward, high-throughput...

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Main Authors: Sang Ho Jeon, Min Chang Kim, Haejun Lee, Ju-Hee Oh, Hyun Seo Kim, Heawon Lee, Taehoon Park, Young-Joo Lee
Format: Article
Language:English
Published: MDPI AG 2025-02-01
Series:Pharmaceutics
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Online Access:https://www.mdpi.com/1999-4923/17/2/273
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author Sang Ho Jeon
Min Chang Kim
Haejun Lee
Ju-Hee Oh
Hyun Seo Kim
Heawon Lee
Taehoon Park
Young-Joo Lee
author_facet Sang Ho Jeon
Min Chang Kim
Haejun Lee
Ju-Hee Oh
Hyun Seo Kim
Heawon Lee
Taehoon Park
Young-Joo Lee
author_sort Sang Ho Jeon
collection DOAJ
description <b>Objectives</b>: Ultrafiltration (UF) is widely accepted as a method for assessing the plasma protein binding (PPB) of drugs. However, it is vulnerable to non-specific binding (NSB) to the device, which can result in inaccuracies. This study presents a straightforward, high-throughput modified UF method aimed at minimizing bias due to NSB. <b>Methods</b>: The modified UF method, sequential UF, features the addition of a 2 min pre-UF phase designed to saturate the NSB in the device, followed by the main 20 min UF procedure, compared to the conventional UF method. To evaluate the feasibility of this sequential UF method, we measured the PPB of nine compounds using sequential UF and compared these results to those obtained with the conventional mass balance UF method, recognized as a standard for NSB correction. <b>Results</b>: The PPB values determined through sequential UF were generally consistent with those derived from the mass balance UF method. The fold differences ranged from 97.9% to 113.8%, with an average of 103.5%. No significant differences were observed between the two methods for all compounds, with the exception of quercetin, which showed an unusually high PPB. <b>Conclusions</b>: Sequential UF was effective in correcting NSB to the device while providing advantages in terms of simplicity and efficiency.
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spelling doaj-art-2581bcdf369b454ea909bc1a67ded89c2025-08-20T03:12:19ZengMDPI AGPharmaceutics1999-49232025-02-0117227310.3390/pharmaceutics17020273A Sequential Ultrafiltration Method to Enhance the Accuracy and Throughput in Plasma Protein Binding TestsSang Ho Jeon0Min Chang Kim1Haejun Lee2Ju-Hee Oh3Hyun Seo Kim4Heawon Lee5Taehoon Park6Young-Joo Lee7Department of Biomedical and Pharmaceutical Sciences, Kyung Hee University, 1 Hoegi-dong, Dongdaemun-gu, Seoul 130-701, Republic of KoreaDepartment of Biomedical and Pharmaceutical Sciences, Kyung Hee University, 1 Hoegi-dong, Dongdaemun-gu, Seoul 130-701, Republic of KoreaDepartment of Biomedical and Pharmaceutical Sciences, Kyung Hee University, 1 Hoegi-dong, Dongdaemun-gu, Seoul 130-701, Republic of KoreaDivision of Biopharmaceutics, College of Pharmacy, Kyung Hee University, 1 Hoegi-dong, Dongdaemun-gu, Seoul 130-701, Republic of KoreaDivision of Biopharmaceutics, College of Pharmacy, Kyung Hee University, 1 Hoegi-dong, Dongdaemun-gu, Seoul 130-701, Republic of KoreaDepartment of Biomedical and Pharmaceutical Sciences, Kyung Hee University, 1 Hoegi-dong, Dongdaemun-gu, Seoul 130-701, Republic of KoreaDepartment of Biomedical and Pharmaceutical Sciences, Kyung Hee University, 1 Hoegi-dong, Dongdaemun-gu, Seoul 130-701, Republic of KoreaDivision of Biopharmaceutics, College of Pharmacy, Kyung Hee University, 1 Hoegi-dong, Dongdaemun-gu, Seoul 130-701, Republic of Korea<b>Objectives</b>: Ultrafiltration (UF) is widely accepted as a method for assessing the plasma protein binding (PPB) of drugs. However, it is vulnerable to non-specific binding (NSB) to the device, which can result in inaccuracies. This study presents a straightforward, high-throughput modified UF method aimed at minimizing bias due to NSB. <b>Methods</b>: The modified UF method, sequential UF, features the addition of a 2 min pre-UF phase designed to saturate the NSB in the device, followed by the main 20 min UF procedure, compared to the conventional UF method. To evaluate the feasibility of this sequential UF method, we measured the PPB of nine compounds using sequential UF and compared these results to those obtained with the conventional mass balance UF method, recognized as a standard for NSB correction. <b>Results</b>: The PPB values determined through sequential UF were generally consistent with those derived from the mass balance UF method. The fold differences ranged from 97.9% to 113.8%, with an average of 103.5%. No significant differences were observed between the two methods for all compounds, with the exception of quercetin, which showed an unusually high PPB. <b>Conclusions</b>: Sequential UF was effective in correcting NSB to the device while providing advantages in terms of simplicity and efficiency.https://www.mdpi.com/1999-4923/17/2/273plasma protein bindingultrafiltrationnon-specific binding
spellingShingle Sang Ho Jeon
Min Chang Kim
Haejun Lee
Ju-Hee Oh
Hyun Seo Kim
Heawon Lee
Taehoon Park
Young-Joo Lee
A Sequential Ultrafiltration Method to Enhance the Accuracy and Throughput in Plasma Protein Binding Tests
Pharmaceutics
plasma protein binding
ultrafiltration
non-specific binding
title A Sequential Ultrafiltration Method to Enhance the Accuracy and Throughput in Plasma Protein Binding Tests
title_full A Sequential Ultrafiltration Method to Enhance the Accuracy and Throughput in Plasma Protein Binding Tests
title_fullStr A Sequential Ultrafiltration Method to Enhance the Accuracy and Throughput in Plasma Protein Binding Tests
title_full_unstemmed A Sequential Ultrafiltration Method to Enhance the Accuracy and Throughput in Plasma Protein Binding Tests
title_short A Sequential Ultrafiltration Method to Enhance the Accuracy and Throughput in Plasma Protein Binding Tests
title_sort sequential ultrafiltration method to enhance the accuracy and throughput in plasma protein binding tests
topic plasma protein binding
ultrafiltration
non-specific binding
url https://www.mdpi.com/1999-4923/17/2/273
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