Skip to main content

Lund University Publications

LUND UNIVERSITY LIBRARIES

Characterization of binding between model protein GA-Z and human serum albumin using asymmetrical flow field-flow fractionation and small angle X-ray scattering

Choi, Jaeyeong LU orcid ; Wahlgren, Marie LU orcid ; Ek, Vilhelm ; Elofsson, Ulla ; Fransson, Jonas LU ; Nilsson, Lars LU ; Terry, Ann LU and Söderberg, Christopher A.G. LU (2020) In PLoS ONE 15(11 November).
Abstract

Protein-based drugs often require targeted drug delivery for optimal therapy. A successful strategy to increase the circulation time of the protein in the blood is to link the therapeutic protein with an albumin-binding domain. In this work, we characterized such a proteinbased drug, GA-Z. Using asymmetrical flow field-flow fractionation coupled with multi-angle light scattering (AF4-MALS) we investigated the GA-Z monomer-dimer equilibrium as well as the molar binding ratio of GA-Z to HSA. Using small angle X-ray scattering, we studied the structure of GA-Z as well as the complex between GA-Z and HSA. The results show that GA-Z is predominantly dimeric in solution at pH 7 and that it binds to monomeric as well as dimeric HSA.... (More)

Protein-based drugs often require targeted drug delivery for optimal therapy. A successful strategy to increase the circulation time of the protein in the blood is to link the therapeutic protein with an albumin-binding domain. In this work, we characterized such a proteinbased drug, GA-Z. Using asymmetrical flow field-flow fractionation coupled with multi-angle light scattering (AF4-MALS) we investigated the GA-Z monomer-dimer equilibrium as well as the molar binding ratio of GA-Z to HSA. Using small angle X-ray scattering, we studied the structure of GA-Z as well as the complex between GA-Z and HSA. The results show that GA-Z is predominantly dimeric in solution at pH 7 and that it binds to monomeric as well as dimeric HSA. Furthermore, GA-Z binds to HSA both as a monomer and a dimer, and thus, it can be expected to stay bound also upon dilution following injection in the blood stream. The results from SAXS and binding studies indicate that the GA-Z dimer is formed between two target domains (Z-domains). The results also indicate that the binding of GA-Z to HSA does not affect the ratio between HSA dimers and monomers, and that no higher order oligomers of the complex are seen other than those containing dimers of GA-Z and dimers of HSA.

(Less)
Please use this url to cite or link to this publication:
author
; ; ; ; ; ; and
organization
publishing date
type
Contribution to journal
publication status
published
subject
in
PLoS ONE
volume
15
issue
11 November
article number
e0242605
pages
19 pages
publisher
Public Library of Science (PLoS)
external identifiers
  • scopus:85093704062
  • pmid:33232370
ISSN
1932-6203
DOI
10.1371/journal.pone.0242605
language
English
LU publication?
yes
id
05017df6-3404-47a8-ac95-14c3c15ea508
date added to LUP
2020-12-16 18:30:04
date last changed
2024-01-17 18:11:09
@article{05017df6-3404-47a8-ac95-14c3c15ea508,
  abstract     = {{<p>Protein-based drugs often require targeted drug delivery for optimal therapy. A successful strategy to increase the circulation time of the protein in the blood is to link the therapeutic protein with an albumin-binding domain. In this work, we characterized such a proteinbased drug, GA-Z. Using asymmetrical flow field-flow fractionation coupled with multi-angle light scattering (AF4-MALS) we investigated the GA-Z monomer-dimer equilibrium as well as the molar binding ratio of GA-Z to HSA. Using small angle X-ray scattering, we studied the structure of GA-Z as well as the complex between GA-Z and HSA. The results show that GA-Z is predominantly dimeric in solution at pH 7 and that it binds to monomeric as well as dimeric HSA. Furthermore, GA-Z binds to HSA both as a monomer and a dimer, and thus, it can be expected to stay bound also upon dilution following injection in the blood stream. The results from SAXS and binding studies indicate that the GA-Z dimer is formed between two target domains (Z-domains). The results also indicate that the binding of GA-Z to HSA does not affect the ratio between HSA dimers and monomers, and that no higher order oligomers of the complex are seen other than those containing dimers of GA-Z and dimers of HSA.</p>}},
  author       = {{Choi, Jaeyeong and Wahlgren, Marie and Ek, Vilhelm and Elofsson, Ulla and Fransson, Jonas and Nilsson, Lars and Terry, Ann and Söderberg, Christopher A.G.}},
  issn         = {{1932-6203}},
  language     = {{eng}},
  number       = {{11 November}},
  publisher    = {{Public Library of Science (PLoS)}},
  series       = {{PLoS ONE}},
  title        = {{Characterization of binding between model protein GA-Z and human serum albumin using asymmetrical flow field-flow fractionation and small angle X-ray scattering}},
  url          = {{http://dx.doi.org/10.1371/journal.pone.0242605}},
  doi          = {{10.1371/journal.pone.0242605}},
  volume       = {{15}},
  year         = {{2020}},
}