Construction of Chimeric Dual-Chain Avidin by Tandem Fusion of the Related Avidins

Tiina A. Riihimäki, Sampo Kukkurainen, Suvi Varjonen, Jarno Hörhä, Thomas K.M. Nyholm, Markku S. Kulomaa, Vesa P. Hytönen (Corresponding Author)

Research output: Contribution to journalArticleScientificpeer-review

6 Citations (Scopus)



Avidin is a chicken egg-white protein with high affinity to vitamin H, also known as D-biotin. Many applications in life science research are based on this strong interaction. Avidin is a homotetrameric protein, which promotes its modification to symmetrical entities. Dual-chain avidin, a genetically engineered avidin form, has two circularly permuted chicken avidin monomers that are tandem-fused into one polypeptide chain. This form of avidin enables independent modification of the two domains, including the two biotin-binding pockets; however, decreased yields in protein production, compared to wt avidin, and complicated genetic manipulation of two highly similar DNA sequences in the tandem gene have limited the use of dual-chain avidin in biotechnological applications.

Principal Findings

To overcome challenges associated with the original dual-chain avidin, we developed chimeric dual-chain avidin, which is a tandem fusion of avidin and avidin-related protein 4 (AVR4), another member of the chicken avidin gene family. We observed an increase in protein production and better thermal stability, compared with the original dual-chain avidin. Additionally, PCR amplification of the hybrid gene was more efficient, thus enabling more convenient and straightforward modification of the dual-chain avidin. When studied closer, the generated chimeric dual-chain avidin showed biphasic biotin dissociation.


The improved dual-chain avidin introduced here increases its potential for future applications. This molecule offers a valuable base for developing bi-functional avidin tools for bioseparation, carrier proteins, and nanoscale adapters. Additionally, this strategy could be helpful when generating hetero-oligomers from other oligomeric proteins with high structural similarity.

Original languageEnglish
Article numbere20535
Number of pages10
JournalPLoS ONE
Issue number5
Publication statusPublished - 2011
MoE publication typeA1 Journal article-refereed


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