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Validation of a Double-Sandwich Enzyme-Linked Immunoassay for Pharmacokinetic Study of an rh-aFGF Hydrogel in Rat Skin and Serum

In this study, we validated a double-sandwich enzyme-linked immunosorbent assay (ELISA) to investigate the pharmacokinetics of a recombinant human acidic fibroblast growth factor (rh-aFGF) hydrogel in rat skin and serum. A total of 130 Sprague-Dawley rats were divided into a control group, rh-aFGF h...

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Autores principales: Hui, Qi, Yang, Rongshuai, Lu, Chao, Bi, Jianing, Li, Lijia, Gong, Jianxiang, Zhang, Li, Jin, Zi, Li, Xiaokun, Wang, Xiaojie
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7248203/
https://www.ncbi.nlm.nih.gov/pubmed/32508643
http://dx.doi.org/10.3389/fphar.2020.00700
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author Hui, Qi
Yang, Rongshuai
Lu, Chao
Bi, Jianing
Li, Lijia
Gong, Jianxiang
Zhang, Li
Jin, Zi
Li, Xiaokun
Wang, Xiaojie
author_facet Hui, Qi
Yang, Rongshuai
Lu, Chao
Bi, Jianing
Li, Lijia
Gong, Jianxiang
Zhang, Li
Jin, Zi
Li, Xiaokun
Wang, Xiaojie
author_sort Hui, Qi
collection PubMed
description In this study, we validated a double-sandwich enzyme-linked immunosorbent assay (ELISA) to investigate the pharmacokinetics of a recombinant human acidic fibroblast growth factor (rh-aFGF) hydrogel in rat skin and serum. A total of 130 Sprague-Dawley rats were divided into a control group, rh-aFGF hydrogel group, and a positive-control group (commercial rh-aFGF-Ai). We first determined the dilution ratio of skin homogenate and then validated the quantitative range, specificity, precision, and accuracy of our double-sandwich ELISA method, as well as the stability of our rh-aFGF hydrogel. For our pharmacokinetic study, skin and serum samples were collected at 0.5, 1, 2, 4, 6, and 10 h after rh-aFGF administration, and the concentration of rh-aFGF was measured by ELISA. The results showed that a 10-fold dilution of the skin tissue homogenate circumvented non-specific interference with endogenous proteins. The quantitative scope of the rh-aFGF calibration curve ranged from 62.5 to 4,000 pg/mL. The precision and accuracy of rh-aFGF quality-control samples were below 20%. Furthermore, bFGF, FGF21, KGF-2, and insulin did not interfere with the detection of aFGF, confirming that our method was specific. Rh-aFGF was stable under normal storage conditions. The maximum concentration (C(max)) and time to peak (T(max)) of the rh-aFGF hydrogel were 909.2 pg/cm(2) and 0.5 h, respectively. The relative bioavailability (F) of the rh-aFGF hydrogel was 120% compared with that of rh-aFGF-Ai. The serum concentration of rh-aFGF was too low to be detected. Taken together, the pharmacokinetics of this rh-aFGF hydrogel provide further support for clinical research on rh-aFGF, and our double-sandwich ELISA method may be useful for pharmacokinetic studies of other protein-based drugs.
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spelling pubmed-72482032020-06-05 Validation of a Double-Sandwich Enzyme-Linked Immunoassay for Pharmacokinetic Study of an rh-aFGF Hydrogel in Rat Skin and Serum Hui, Qi Yang, Rongshuai Lu, Chao Bi, Jianing Li, Lijia Gong, Jianxiang Zhang, Li Jin, Zi Li, Xiaokun Wang, Xiaojie Front Pharmacol Pharmacology In this study, we validated a double-sandwich enzyme-linked immunosorbent assay (ELISA) to investigate the pharmacokinetics of a recombinant human acidic fibroblast growth factor (rh-aFGF) hydrogel in rat skin and serum. A total of 130 Sprague-Dawley rats were divided into a control group, rh-aFGF hydrogel group, and a positive-control group (commercial rh-aFGF-Ai). We first determined the dilution ratio of skin homogenate and then validated the quantitative range, specificity, precision, and accuracy of our double-sandwich ELISA method, as well as the stability of our rh-aFGF hydrogel. For our pharmacokinetic study, skin and serum samples were collected at 0.5, 1, 2, 4, 6, and 10 h after rh-aFGF administration, and the concentration of rh-aFGF was measured by ELISA. The results showed that a 10-fold dilution of the skin tissue homogenate circumvented non-specific interference with endogenous proteins. The quantitative scope of the rh-aFGF calibration curve ranged from 62.5 to 4,000 pg/mL. The precision and accuracy of rh-aFGF quality-control samples were below 20%. Furthermore, bFGF, FGF21, KGF-2, and insulin did not interfere with the detection of aFGF, confirming that our method was specific. Rh-aFGF was stable under normal storage conditions. The maximum concentration (C(max)) and time to peak (T(max)) of the rh-aFGF hydrogel were 909.2 pg/cm(2) and 0.5 h, respectively. The relative bioavailability (F) of the rh-aFGF hydrogel was 120% compared with that of rh-aFGF-Ai. The serum concentration of rh-aFGF was too low to be detected. Taken together, the pharmacokinetics of this rh-aFGF hydrogel provide further support for clinical research on rh-aFGF, and our double-sandwich ELISA method may be useful for pharmacokinetic studies of other protein-based drugs. Frontiers Media S.A. 2020-05-19 /pmc/articles/PMC7248203/ /pubmed/32508643 http://dx.doi.org/10.3389/fphar.2020.00700 Text en Copyright © 2020 Hui, Yang, Lu, Bi, Li, Gong, Zhang, Jin, Li and Wang http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Pharmacology
Hui, Qi
Yang, Rongshuai
Lu, Chao
Bi, Jianing
Li, Lijia
Gong, Jianxiang
Zhang, Li
Jin, Zi
Li, Xiaokun
Wang, Xiaojie
Validation of a Double-Sandwich Enzyme-Linked Immunoassay for Pharmacokinetic Study of an rh-aFGF Hydrogel in Rat Skin and Serum
title Validation of a Double-Sandwich Enzyme-Linked Immunoassay for Pharmacokinetic Study of an rh-aFGF Hydrogel in Rat Skin and Serum
title_full Validation of a Double-Sandwich Enzyme-Linked Immunoassay for Pharmacokinetic Study of an rh-aFGF Hydrogel in Rat Skin and Serum
title_fullStr Validation of a Double-Sandwich Enzyme-Linked Immunoassay for Pharmacokinetic Study of an rh-aFGF Hydrogel in Rat Skin and Serum
title_full_unstemmed Validation of a Double-Sandwich Enzyme-Linked Immunoassay for Pharmacokinetic Study of an rh-aFGF Hydrogel in Rat Skin and Serum
title_short Validation of a Double-Sandwich Enzyme-Linked Immunoassay for Pharmacokinetic Study of an rh-aFGF Hydrogel in Rat Skin and Serum
title_sort validation of a double-sandwich enzyme-linked immunoassay for pharmacokinetic study of an rh-afgf hydrogel in rat skin and serum
topic Pharmacology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7248203/
https://www.ncbi.nlm.nih.gov/pubmed/32508643
http://dx.doi.org/10.3389/fphar.2020.00700
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