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Characterization of binding between model protein GA-Z and human serum albumin using asymmetrical flow field-flow fractionation and small angle X-ray scattering.
Lund University, Sweden.
Lund University, Sweden.
Swedish Orphan Biovitrum AB, Sweden.
RISE Research Institutes of Sweden, Bioeconomy and Health, Chemical Process and Pharmaceutical Development.ORCID iD: 0000-0002-3350-0242
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2020 (English)In: PLOS ONE, E-ISSN 1932-6203, Vol. 15, no 11, article id e0242605Article in journal (Refereed) Published
Abstract [en]

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 protein-based 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.

Place, publisher, year, edition, pages
2020. Vol. 15, no 11, article id e0242605
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Natural Sciences
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URN: urn:nbn:se:ri:diva-50945DOI: 10.1371/journal.pone.0242605PubMedID: 33232370OAI: oai:DiVA.org:ri-50945DiVA, id: diva2:1506270
Available from: 2020-12-02 Created: 2020-12-02 Last updated: 2023-12-07Bibliographically approved

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Elofsson, Ulla

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