Phenotype-specific recombinant haptoglobin polymers co-expressed with C1r-like protein as optimized hemoglobin-binding therapeutics
dc.contributor.author
Schaer, Christian A.
dc.contributor.author
Owczarek, Catherine
dc.contributor.author
Deuel, Jeremy W.
dc.contributor.author
Schauer, Stefan
dc.contributor.author
Baek, Jin Hyen
dc.contributor.author
Yalamanoglu, Ayla
dc.contributor.author
Hardy, Matthew P.
dc.contributor.author
Scotney, Pierre D.
dc.contributor.author
Schmidt, Peter M.
dc.contributor.author
Pelzing, Matthias
dc.contributor.author
Soupourmas, Peter
dc.contributor.author
Buehler, Paul W.
dc.contributor.author
Schaer, Dominik J.
dc.date.accessioned
2019-01-23T17:43:34Z
dc.date.available
2019-01-23T08:17:16Z
dc.date.available
2019-01-23T17:43:34Z
dc.date.issued
2018-03-15
dc.identifier.issn
1472-6750
dc.identifier.other
10.1186/s12896-018-0424-3
en_US
dc.identifier.uri
http://hdl.handle.net/20.500.11850/318753
dc.identifier.doi
10.3929/ethz-b-000318753
dc.description.abstract
Background
Preclinical studies have evaluated haptoglobin (Hp) polymers from pooled human plasma as a therapeutic protein to attenuate toxic effects of cell-free hemoglobin (Hb). Proof of concept studies have demonstrated efficacy of Hp in hemolysis associated with transfusion and sickle cell anemia. However, phenotype-specific Hp products might be desirable to exploit phenotype specific activities of Hp 1–1 versus Hp 2–2, offering opportunities for recombinant therapeutics. Prohaptoglobin (proHp) is the primary translation product of the Hp mRNA. ProHp is proteolytically cleaved by complement C1r subcomponent-like protein (C1r-LP) in the endoplasmic reticulum. Two main allelic Hp variants, HP1 and HP2 exist. The larger HP2 is considered to be the ancestor variant of all human Hp alleles and is characterized by an α2-chain, which contains an extra cysteine residue that pairs with additional α-chains generating multimers with molecular weights of 200–900 kDa. The two human HP1 alleles (HP1F and HP1S) differ by a two-amino-acid substitution polymorphism within the α-chain and are derived from HP2 by recurring exon deletions.
Results
In the present study, we describe a process for the production of recombinant phenotype specific Hp polymers in mammalian FS293F cells. This approach demonstrates that efficient expression of mature and fully functional protein products requires co-expression of active C1r-LP. The functional characterization of our proteins, which included monomer/polymer distribution, binding affinities as well as NO-sparing and antioxidant functions, demonstrated that C1r-LP-processed recombinant Hp demonstrates equal protective functions as plasma derived Hp in vitro as well as in animal studies.
Conclusions
We present a recombinant production process for fully functional phenotype-specific Hp therapeutics. The proposed process could accelerate the development of Hb scavengers to treat patients with cell-free Hb associated disease states, such as sickle cell disease and other hemolytic conditions.
en_US
dc.format
application/pdf
en_US
dc.language.iso
en
en_US
dc.publisher
BioMed Central
dc.rights.uri
http://creativecommons.org/licenses/by/4.0/
dc.title
Phenotype-specific recombinant haptoglobin polymers co-expressed with C1r-like protein as optimized hemoglobin-binding therapeutics
en_US
dc.type
Journal Article
dc.rights.license
Creative Commons Attribution 4.0 International
ethz.journal.title
BMC Biotechnology
ethz.journal.volume
18
en_US
ethz.journal.issue
1
en_US
ethz.journal.abbreviated
BMC biotechnol. (Online)
ethz.pages.start
15
en_US
ethz.size
13 p.
en_US
ethz.version.deposit
publishedVersion
en_US
ethz.publication.place
London
ethz.publication.status
published
en_US
ethz.leitzahl
ETH Zürich::00002 - ETH Zürich::00003 - Schulleitung und Dienste::00022 - Bereich VP Forschung / Domain VP Research::02207 - Functional Genomics Center Zurich / Functional Genomics Center Zurich
en_US
ethz.leitzahl.certified
ETH Zürich::00002 - ETH Zürich::00003 - Schulleitung und Dienste::00022 - Bereich VP Forschung / Domain VP Research::02207 - Functional Genomics Center Zurich / Functional Genomics Center Zurich
en_US
ethz.date.deposited
2019-01-23T08:17:23Z
ethz.source
FORM
ethz.eth
yes
en_US
ethz.availability
Open access
en_US
ethz.rosetta.installDate
2019-01-23T17:43:49Z
ethz.rosetta.lastUpdated
2024-02-02T07:01:57Z
ethz.rosetta.versionExported
true
ethz.COinS
ctx_ver=Z39.88-2004&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.atitle=Phenotype-specific%20recombinant%20haptoglobin%20polymers%20co-expressed%20with%20C1r-like%20protein%20as%20optimized%20hemoglobin-binding%20therapeutics&rft.jtitle=BMC%20Biotechnology&rft.date=2018-03-15&rft.volume=18&rft.issue=1&rft.spage=15&rft.issn=1472-6750&rft.au=Schaer,%20Christian%20A.&Owczarek,%20Catherine&Deuel,%20Jeremy%20W.&Schauer,%20Stefan&Baek,%20Jin%20Hyen&rft.genre=article&rft_id=info:doi/10.1186/s12896-018-0424-3&
Files in this item
Publication type
-
Journal Article [133598]