Change search
ReferencesLink to record
Permanent link

Direct link
Genetic fusion of single-chain variable fragments to partial spider silk improves target detection in micro- and nanoarrays
Show others and affiliations
2016 (English)In: Biotechnology Journal, ISSN 1860-6768, E-ISSN 1860-7314, Vol. 11, no 3Article in journal (Refereed) PublishedText
Abstract [en]

Immobilizing biomolecules with retained functionality and stability on solid supports is crucial for generation of sensitive immunoassays. However, upon use of conventional immobilization strategies, a major portion of the biomolecules (e.g. antibodies) frequently tends to lose their bioactivity. In this study, we describe a procedure to immobilize human single-chain variable fragment (scFv) via genetic fusion to partial spider silk, which have a high tendency to adhere to solid supports. Two scFvs, directed towards serum proteins, were genetically fused to partial spider silk proteins and expressed as silk fusion proteins in E. coli. Antigen binding ability of scFvs attached to a partial silk protein denoted RC was investigated using microarray analysis, whereas scFvs fused to the NC silk variant were examined using nanoarrays. Results from micro- and nanoarrays confirmed the functionality of scFvs attached to both RC and NC silk, and also for binding of targets in crude serum. Furthermore, the same amount of added scFv gives higher signal intensity when immobilized via partial spider silk compared to when immobilized alone. Together, the results suggest that usage of scFv-silk fusion proteins in immunoassays could improve target detection, in the long run enabling novel biomarkers to be detected in crude serum proteomes.

Place, publisher, year, edition, pages
Wiley-VCH Verlagsgesellschaft, 2016. Vol. 11, no 3
Keyword [en]
Antigen detection, Microarray, Nanoarray, Single-chain variable fragment, Spider silk
National Category
Biochemistry and Molecular Biology
Identifiers
URN: urn:nbn:se:kth:diva-185068DOI: 10.1002/biot.201500297ISI: 000372141400016PubMedID: 26470853ScopusID: 2-s2.0-84959232625OAI: oai:DiVA.org:kth-185068DiVA: diva2:920048
Note

QC 20160415

Available from: 2016-04-15 Created: 2016-04-11 Last updated: 2016-04-15Bibliographically approved

Open Access in DiVA

No full text

Other links

Publisher's full textPubMedScopus

Search in DiVA

By author/editor
Hedhammar, My
By organisation
Protein Technology
In the same journal
Biotechnology Journal
Biochemistry and Molecular Biology

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: 8 hits
ReferencesLink to record
Permanent link

Direct link