In vitro transcytosis assay
AssigneeGenentech, Inc.
InventorsShan CHUNG, Van Hong NGUYEN, An SONG
Life of this patent
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Abstract
The present disclosure relates to methods and compositions useful for measuring the transcytosis of a molecule. In particular, the present disclosure relates to in vitro receptor-dependent transcytosis assays for evaluating the clearance rates of therapeutic antibody molecules and Fc-fusion proteins in humans and animals.
Claims54
A method for determining a pharmacokinetic (PK) parameter of a molecule, the method comprising determining a measure of transcytosis of the molecule or a plurality of the molecules through a cell or a plurality of cells under conditions of physiological pH.
A method for measuring the transcellular transportation of a plurality of a single molecule or a plurality of distinct molecules, comprising:
- a) introducing the plurality of a single molecule or the plurality of distinct molecules into a first of two chambers where the first chamber is separated from the second chamber by a cell or plurality of cells, and wherein each of the first and second chambers has a physiological pH value;
- and
- b) measuring the number of molecules transcytosed from the first chamber to the second chamber.
The method of claim 21, wherein the at least one heterologous gene is selected from the group consisting of a FCGRT gene and a B2M gene;
- and/or the heterologous cell surface protein comprises a neonatal Fc receptor (FcRn).
A method of determining the in vivo clearance of a molecule, comprising:
- a) introducing the molecule into a first of two chambers where the first chamber is separated from the second chamber by a cell or plurality of cells, and wherein each of the first and second chambers has a physiological pH value;
- b) measuring the number of molecules transcytosed from the first chamber to the second chamber;
- and
- c) determining the in vivo clearance of the molecule based on the number of molecules measured.
An assay for measuring the transcellular transportation of a plurality of molecules, comprising a first chamber and a second chamber, each of the first and second chambers having a physiological pH value, wherein the first chamber receives a plurality of molecules, is configured to transcellularly transport the molecules to the second chamber, and the transcellular transportation of the plurality of molecules is measured.
An assay system comprising:
- a) a first chamber and a second chamber, wherein each of the first and second chambers has a physiological pH;
- b) a monolayer of cells positioned in the first chamber such that the cells can mediate the transcytosis of a molecule introduced into the first chamber to the second chamber;
- and
- c) a detector for detecting the presence of a molecule in the second chamber.
Dependencies are inferred from the claim text.