A novel in vitro assay to predict neonatal Fc receptor-mediated human IgG half-life

MAbs. 2015;7(5):912-21. doi: 10.1080/19420862.2015.1054585.


Immunoglobulin G (IgG) has an unusually long serum half-life in comparison to proteins of a similar size. It is well-known that this phenomenon is due to IgG's ability to bind the neonatal Fc receptor (FcRn) in a pH-dependent manner. FcRn binding properties can vary among IgGs, resulting in altered in vivo half-lives, and therefore it would be beneficial to accurately predict the FcRn binding properties of therapeutic IgG monoclonal antibodies (mAbs). Here we describe the development of an in vitro model capable of predicting the in vivo half-life of human IgG. Using a high-throughput biolayer interferometry (BLI) platform, the human FcRn association rate at acidic pH and subsequent dissociation rate at physiological pH was determined for 5 human IgG1 mAbs. Comparing the combined FcRn association and dissociation rates to the Phase 1 clinical study half-lives of the mAbs resulted in a strong correlation. The correlation was also verified in vivo using mice transgenic for human FcRn. The model was used to characterize various factors that may influence FcRn-mAb binding, including mAb variable region sequence differences and constant region glycosylation patterns. Results indicated that the complementarity-determining regions of the heavy chain significantly influence the mAb's FcRn binding properties, while the absence of glycosylation does not alter mAb-FcRn binding. Development of this high-throughput FcRn binding model could potentially predict the half-life of therapeutic IgGs and aid in selection of lead candidates while also serving as a screening tool for the development of mAbs with desired pharmacokinetic properties.

Keywords: IgG monoclonal antibody; biolayer interferometry; correlation; glycosylation; half-life; in vitro; mutations; neonatal Fc receptor (FcRn); pharmacokinetics (PK); transgenic mice.

Publication types

  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Animals
  • Antibodies, Monoclonal / blood*
  • Blotting, Western
  • Electrophoresis, Polyacrylamide Gel
  • Half-Life
  • Histocompatibility Antigens Class I / metabolism*
  • Humans
  • Immunoglobulin G / blood*
  • In Vitro Techniques
  • Mice
  • Mice, Transgenic
  • Receptors, Fc / metabolism*


  • Antibodies, Monoclonal
  • Histocompatibility Antigens Class I
  • Immunoglobulin G
  • Receptors, Fc
  • Fc receptor, neonatal