Half-life extension of a single-chain diabody by fusion to domain B of staphylococcal protein A

  • Unverdorben F
  • Färber-Schwarz A
  • Richter F
 et al. 
  • 49

    Readers

    Mendeley users who have this article in their library.
  • 10

    Citations

    Citations of this article.

Abstract

Binding of a therapeutic protein to a long-circulating plasma protein can result in a strongly extended half-life. Among these plasma proteins, albumin and immunoglobulins are of special interest because of their exceptionally long half-life, which is to a great extent determined by recycling through the neonatal Fc receptor (FcRn). Many strategies have been established employing reversible binding to albumin, e.g. using an albumin-binding domain from streptococcal protein G. We show here that the half-life of a recombinant antibody molecule can also be prolonged by fusion to a single immunoglobulin-binding domain (IgBD) from staphylococcal protein A. This domain (domain B, SpAB) is composed of 56 amino acid residues and was fused to the C-terminus of a bispecific single-chain diabody (scDb). The scDb-SpAB fusion protein was produced in HEK293 cells and retained its antigen-binding activity as shown by enzyme-linked immunosorbent assay and flow cytometry. Furthermore, the fusion protein was capable of binding to human and mouse IgG in a pH-dependent manner. In mice, the terminal half-life of the fusion protein was improved from ∼1–2 h of the unmodified scDb to 11.8 h. Although the fusion protein did not reach the long half-life seen for IgG, our results established the applicability of a single bacterial IgBD for half-life extension purposes.

Author-supplied keywords

  • immunoglobulin-binding domain
  • pharmacokinetics
  • protein A
  • serum half-life
  • single-chain diabody

Get free article suggestions today

Mendeley saves you time finding and organizing research

Sign up here
Already have an account ?Sign in

Find this document

Get full text

Authors

  • Felix Unverdorben

  • Aline Färber-Schwarz

  • Fabian Richter

  • Meike Hutt

  • Roland E. Kontermann

Cite this document

Choose a citation style from the tabs below

Save time finding and organizing research with Mendeley

Sign up for free