Vitro prediction of in vivo half-life

Patent metadata

Field	Value
Publication number	US-12461109-B2
Application number	US-202117158431-A
Country	US
Kind code	B2
Filing date	Jan 26, 2021
Priority date	Mar 21, 2014
Publication date	Nov 4, 2025
Grant date	Nov 4, 2025

Abstract

Herein is reported a method for determining the presence of antibody-Fab-FcRn interaction in an antibody-Fc-FcRn complex influencing the in vivo half-life comprising the steps of a) determining the retention time of the antibody on an FcRn affinity chromatography column with a positive linear pH gradient elution in the presence of a first sodium chloride concentration, and b) determining the retention time of the antibody on an FcRn affinity chromatography column with a positive linear pH gradient elution in the presence of a second sodium chloride concentration, whereby the presence of antibody-Fab-FcRn interaction in an antibody-Fc-FcRn complex influencing the in vivo half-life is determined if the retention time determined in step a) and the retention time determined in step b) are substantially different.

First claim

What is claimed is:

1 . A method for selecting a variant antibody of a parent antibody, comprising the steps of:
determining a first retention time of the variant antibody and the parent antibody on an FcRn affinity chromatography column with a positive linear pH gradient elution in the presence of a first salt concentration, and determining a second retention time of the variant antibody on an FcRn affinity chromatography column with the positive linear pH gradient elution in the presence of a second salt concentration, wherein:
the method is for selecting the variant antibody of the IgG1, IgG3 or IgG4 subclass that has a relative in vivo half-life that is increased compared to the parent antibody of the IgG1, IgG3 or IgG4 subclass, by selecting a variant antibody that has a first retention time that is longer than the first retention time of the parent antibody, and a first retention time that is substantially the same as the second retention time,

wherein the positive linear pH gradient is from about pH 5.5 to about pH 8.8; the salt is sodium chloride; and
wherein the first salt concentration is about 140 mM; and wherein the second salt concentration is about 400 mM.

2 . The method according to claim 1 , wherein the method is for selecting a variant antibody that is free of antibody-Fab-FcRn interaction influencing the in vivo half-life of the antibody.

3 . The method according to claim 1 , wherein substantially same retention times differ by 3.5% or less.

4 . The method according to claim 1 , wherein the variant antibody is a full length antibody.

Assignees

• Hoffmann La Roche

Inventors

• Emrich Thomas
• Kettenberger Hubert
• Schlothauer Tilman
• Schoch Angela

Classifications

• C07K2317/52

  Constant or Fc region; Isotype · CPC title

• C07K2317/94

  Stability, e.g. half-life, pH, temperature or enzyme-resistance · CPC title

• C07K16/00 Primary

  Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies · CPC title

• B01D15/168 Primary

  pH gradient or chromatofocusing, i.e. separation according to the isoelectric point pI · CPC title

• B01D15/3809 Primary

  of the antigen-antibody type, e.g. protein A, G or L chromatography · CPC title