Characterization of drug-protein binding process by employing equilibrium sampling through hollow-fiber supported liquid membrane and Bjerrum and Scatchard plots.
(2008) In Journal of Pharmaceutical and Biomedical Analysis 48(1). p.49-56- Abstract
- The technique equilibrium sampling through membrane (ESTM) was extended to measuring the free drug concentration in solutions of drug and protein. Bjerrum and Scatchard plots were employed for characterizing individual drug binding to pure human blood proteins. Four drugs were investigated as a model system: fluvoxamine and ropivacaine which dominantly bind to alpha-acid glycoprotein (AGP), and R,S-ibuprofen and S-ketoprofen which highly bind to human serum albumin (HSA). The level of drug binding to AGP and HSA relied on drug and protein concentrations. Bjerrum and Scatchard plots revealed high affinity constants (K(a)) at low protein concentration. Both Bjerrum and Scatchard plots of fluvoxamine and ropivacaine binding to AGP showed one... (More)
- The technique equilibrium sampling through membrane (ESTM) was extended to measuring the free drug concentration in solutions of drug and protein. Bjerrum and Scatchard plots were employed for characterizing individual drug binding to pure human blood proteins. Four drugs were investigated as a model system: fluvoxamine and ropivacaine which dominantly bind to alpha-acid glycoprotein (AGP), and R,S-ibuprofen and S-ketoprofen which highly bind to human serum albumin (HSA). The level of drug binding to AGP and HSA relied on drug and protein concentrations. Bjerrum and Scatchard plots revealed high affinity constants (K(a)) at low protein concentration. Both Bjerrum and Scatchard plots of fluvoxamine and ropivacaine binding to AGP showed one specific binding site (n(1)=1) with ropivacaine K(a) value close to 5 times higher than the K(a) of fluvoxamine at 22.9muM AGP concentration. Bjerrum plots of ketoprofen and ibuprofen gave total number of binding sites or bound molecules of 6-7, which did not depend on the drug or protein concentration. Scatchard plots of ketoprofen and ibuprofen exhibited two binding sites (n(1) and n(2)) at 0.15muM and 0.75muM HSA concentrations. On one hand, at 0.15muM HSA, ketoprofen and ibuprofen were bound to site I at n(1)=1.2 and n(1)=1.0, respectively. However, at 0.75muM HSA, ketoprofen and ibuprofen were bound to site I at n(1)=1.2 and n(1)=1.9, respectively. On the other hand, site II, at 0.15muM HSA, interacted with ketoprofen and ibuprofen at n(2)=5.6 and 6.7, respectively. However, at 0.75muM HSA, site II interacted with ketoprofen at n(2)=7.4 and ibuprofen at n(2)=6.2. It would be concluded that, upon mixing ketoprofen and ibuprofen in a HSA solution, a ketoprofen-ibuprofen interaction would most likely occur at site II in HSA. (Less)
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https://lup.lub.lu.se/record/1168677
- author
- Barri, Thaer LU ; Trtić-Petrović, Tatjana ; Karlsson, Michael and Jönsson, Jan Åke LU
- organization
- publishing date
- 2008
- type
- Contribution to journal
- publication status
- published
- subject
- in
- Journal of Pharmaceutical and Biomedical Analysis
- volume
- 48
- issue
- 1
- pages
- 49 - 56
- publisher
- Elsevier
- external identifiers
-
- wos:000259339800007
- pmid:18565712
- scopus:49849105510
- ISSN
- 0731-7085
- DOI
- 10.1016/j.jpba.2008.04.030
- language
- English
- LU publication?
- yes
- additional info
- The information about affiliations in this record was updated in December 2015. The record was previously connected to the following departments: Analytical Chemistry (S/LTH) (011001004)
- id
- f9c00858-4a78-4b07-bc53-7d850620b45c (old id 1168677)
- date added to LUP
- 2016-04-01 11:57:21
- date last changed
- 2022-01-26 20:43:54
@article{f9c00858-4a78-4b07-bc53-7d850620b45c,
abstract = {{The technique equilibrium sampling through membrane (ESTM) was extended to measuring the free drug concentration in solutions of drug and protein. Bjerrum and Scatchard plots were employed for characterizing individual drug binding to pure human blood proteins. Four drugs were investigated as a model system: fluvoxamine and ropivacaine which dominantly bind to alpha-acid glycoprotein (AGP), and R,S-ibuprofen and S-ketoprofen which highly bind to human serum albumin (HSA). The level of drug binding to AGP and HSA relied on drug and protein concentrations. Bjerrum and Scatchard plots revealed high affinity constants (K(a)) at low protein concentration. Both Bjerrum and Scatchard plots of fluvoxamine and ropivacaine binding to AGP showed one specific binding site (n(1)=1) with ropivacaine K(a) value close to 5 times higher than the K(a) of fluvoxamine at 22.9muM AGP concentration. Bjerrum plots of ketoprofen and ibuprofen gave total number of binding sites or bound molecules of 6-7, which did not depend on the drug or protein concentration. Scatchard plots of ketoprofen and ibuprofen exhibited two binding sites (n(1) and n(2)) at 0.15muM and 0.75muM HSA concentrations. On one hand, at 0.15muM HSA, ketoprofen and ibuprofen were bound to site I at n(1)=1.2 and n(1)=1.0, respectively. However, at 0.75muM HSA, ketoprofen and ibuprofen were bound to site I at n(1)=1.2 and n(1)=1.9, respectively. On the other hand, site II, at 0.15muM HSA, interacted with ketoprofen and ibuprofen at n(2)=5.6 and 6.7, respectively. However, at 0.75muM HSA, site II interacted with ketoprofen at n(2)=7.4 and ibuprofen at n(2)=6.2. It would be concluded that, upon mixing ketoprofen and ibuprofen in a HSA solution, a ketoprofen-ibuprofen interaction would most likely occur at site II in HSA.}},
author = {{Barri, Thaer and Trtić-Petrović, Tatjana and Karlsson, Michael and Jönsson, Jan Åke}},
issn = {{0731-7085}},
language = {{eng}},
number = {{1}},
pages = {{49--56}},
publisher = {{Elsevier}},
series = {{Journal of Pharmaceutical and Biomedical Analysis}},
title = {{Characterization of drug-protein binding process by employing equilibrium sampling through hollow-fiber supported liquid membrane and Bjerrum and Scatchard plots.}},
url = {{http://dx.doi.org/10.1016/j.jpba.2008.04.030}},
doi = {{10.1016/j.jpba.2008.04.030}},
volume = {{48}},
year = {{2008}},
}