Fusion protein linkers: property, design and functionality

Adv Drug Deliv Rev. 2013 Oct;65(10):1357-69. doi: 10.1016/j.addr.2012.09.039. Epub 2012 Sep 29.


As an indispensable component of recombinant fusion proteins, linkers have shown increasing importance in the construction of stable, bioactive fusion proteins. This review covers the current knowledge of fusion protein linkers and summarizes examples for their design and application. The general properties of linkers derived from naturally-occurring multi-domain proteins can be considered as the foundation in linker design. Empirical linkers designed by researchers are generally classified into 3 categories according to their structures: flexible linkers, rigid linkers, and in vivo cleavable linkers. Besides the basic role in linking the functional domains together (as in flexible and rigid linkers) or releasing the free functional domain in vivo (as in in vivo cleavable linkers), linkers may offer many other advantages for the production of fusion proteins, such as improving biological activity, increasing expression yield, and achieving desirable pharmacokinetic profiles.

Keywords: Bifunctional recombinant proteins; EBFP; EGFP; FIX; FRET; G-CSF; HSA; IFN; LAP; MMP; PD; PDB; PK; Pharmacodynamics; Pharmacokinetics; Protein Data Bank; Spacer; TGN; Tf; V(H); V(L); aa; amino acid; antibody heavy chain variable region; antibody light-chain variable regions; coagulation factor IX; enhanced blue fluorescent protein; enhanced green fluorescent protein; fluorescent resonance energy transfer; granulocyte colony-stimulating factor; hGH; human growth hormone; human serum albumin; interferon; latency associated peptide; matrix metalloproteinase; pharmacodynamic; pharmacokinetic; scFv; single-chain variable fragment; trans-golgi network; transferrin.

Publication types

  • Review

MeSH terms

  • Animals
  • Drug Delivery Systems*
  • Humans
  • Peptides / chemistry
  • Protein Structure, Tertiary
  • Recombinant Fusion Proteins / administration & dosage
  • Recombinant Fusion Proteins / chemistry*
  • Recombinant Fusion Proteins / metabolism


  • Peptides
  • Recombinant Fusion Proteins