Human Serum Albumin Domain I Fusion Protein for Antibody Conjugation

Bioconjug Chem. 2016 Oct 19;27(10):2271-2275. doi: 10.1021/acs.bioconjchem.6b00432. Epub 2016 Sep 26.

Abstract

Bioorthogonal labeling of antibodies enables the conjugation of compounds, such as small molecules or peptides, which expand targeting capacity or enhance cytotoxicity. Taking advantage of a cyclohexene sulfonamide compound that site-selectively labels Lys64 in human serum albumin (HSA), we demonstrate that domain I of HSA can be used as a fusion protein for the preparation of antibody conjugates. Trastuzumab fusions were expressed at the N-terminus of the light chain or the C-terminus of the heavy chain enabling conjugation to small molecules. Moreover, these conjugates retained HER2 binding and proved to be highly stable in human plasma. Antibody conjugation via HSA domain I fusion should therefore have broad utility for making serum-stable antibody conjugates, particularly for antibody-drug conjugates.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Antibodies / chemistry
  • Breast Neoplasms / metabolism
  • Cell Line, Tumor
  • Female
  • Flow Cytometry
  • Humans
  • Immunoconjugates / blood
  • Immunoconjugates / chemistry*
  • Immunoconjugates / metabolism
  • Lysine / chemistry
  • Protein Domains
  • Protein Engineering / methods
  • Receptor, ErbB-2 / metabolism
  • Recombinant Fusion Proteins / blood
  • Recombinant Fusion Proteins / chemistry*
  • Recombinant Fusion Proteins / metabolism
  • Rhodamines / chemistry
  • Serum Albumin / chemistry*
  • Trastuzumab / chemistry

Substances

  • Antibodies
  • Immunoconjugates
  • Recombinant Fusion Proteins
  • Rhodamines
  • Serum Albumin
  • ERBB2 protein, human
  • Receptor, ErbB-2
  • Lysine
  • Trastuzumab