uu.seUppsala University Publications
Change search
ReferencesLink to record
Permanent link

Direct link
An affibody-adalimumab hybrid blocks combined IL-6 and TNF-triggered serum amyloid A secretion in vivo
Show others and affiliations
2014 (English)In: MABS, ISSN 1942-0862, Vol. 6, no 6, 1598-1607 p.Article in journal (Refereed) Published
Abstract [en]

In inflammatory disease conditions, the regulation of the cytokine system is impaired, leading to tissue damages. Here, we used protein engineering to develop biologicals suitable for blocking a combination of inflammation driving cytokines by a single construct. From a set of interleukin (IL)-6-binding affibody molecules selected by phage display, five variants with a capability of blocking the interaction between complexes of soluble IL-6 receptor a (sIL-6R alpha) and IL6 and the co-receptor gp130 were identified. In cell assays designed to analyze any blocking capacity of the classical or the alternative (trans) signaling IL-6 pathways, one variant, Z(IL-6_13) with an affinity (K-D) for IL-6 of similar to 500 pM, showed the best performance. To construct fusion proteins ("AffiMabs") with dual cytokine specificities, Z(IL-6_13) was fused to either the N-or C-terminus of both the heavy and light chains of the anti-tumor necrosis factor (TNF) monoclonal antibody adalimumab (Humira (R)). One AffiMab construct with Z(IL-6_13) positioned at the N-terminus of the heavy chain, denoted Z(IL-6_13)-HCAda, was determined to be the most optimal, and it was subsequently evaluated in an acute Serum Amyloid A (SAA) model in mice. Administration of the AffiMab or adalimumab prior to challenge with a mix of IL-6 and TNF reduced the levels of serum SAA in a dose-dependent manner. Interestingly, the highest dose (70 mg/kg body weight) of adalimumab only resulted in a 50% reduction of SAA-levels, whereas the corresponding dose of the Z(IL-6_13)-HCAda AffiMab with combined IL-6/TNF specificity, resulted in SAA levels below the detection limit.

Place, publisher, year, edition, pages
2014. Vol. 6, no 6, 1598-1607 p.
Keyword [en]
AffiMab, Antibody, IL-6, TNF, adalimumab, affibody, affinity, fusion, inflammation
National Category
Other Medical Sciences not elsewhere specified
URN: urn:nbn:se:uu:diva-243059DOI: 10.4161/mabs.36089ISI: 000346878600024PubMedID: 25484067OAI: oai:DiVA.org:uu-243059DiVA: diva2:787755
Available from: 2015-02-11 Created: 2015-02-04 Last updated: 2015-02-11Bibliographically approved

Open Access in DiVA

No full text

Other links

Publisher's full textPubMed
By organisation
Department of Radiology, Oncology and Radiation Science
Other Medical Sciences not elsewhere specified

Search outside of DiVA

GoogleGoogle Scholar
The number of downloads is the sum of all downloads of full texts. It may include eg previous versions that are now no longer available

Altmetric score

Total: 173 hits
ReferencesLink to record
Permanent link

Direct link