US 10,520,511 B2Granted

Multimeric protein purity determination

AssigneeRegeneron Pharmaceuticals, Inc.

InventorsKathir Muthusamy, Jiann-Kae Luo

Life of this patent

Estimating the term…

Loading patent…

Abstract

Improved capillary zone electrophoresis (CZE), affinity capillary electrophoresis (ACE), and partially filled-ACE (PF-ACE) systems and methods for the detection and quantification of specific molecular entities in a mixture thereof are provided. During manufacturing, heterodimeric bispecific antibodies are often produced along with homodimer species, which can confound quantification of the bispecific antibody. Disclosed are capillary electrophoretic systems and methods of detecting a specific homodimer in the mixture of bispecific heterodimer and homodimers. A ligand capable of binding one of the subunits of the bispecific antibody is contacted with the mixture to form a complex having a reduced electrophoretic mobility, thereby enabling detection of the unbound homodimer.

Claims18

Dependent claims

A method for separating and detecting a first protein in a mixture of proteins having similar charge to mass ratios, the method comprising:

a. contacting the mixture of proteins including the first protein with a modified ligand that comprises an acidic moiety and does not bind the first protein but does bind other proteins in the mixture of proteins having similar charge to mass ratios to form a sample;
b. applying the sample to a capillary comprising a ligand plug that comprises additional modified ligand;
c. applying a voltage to the capillary to allow the first protein to move through the capillary at a faster rate than the other proteins to separate the first protein from the other proteins;
and
d. detecting the first protein, thereby separating and detecting the first protein in a mixture of proteins having similar charge to mass ratios.

2 more elements

Dependencies are inferred from the claim text.

Facts

Application: 15/152,135Filed 11 May 2016 · us provisional application
Priority: 12 May 2015
Published: US 2017/0045527 A116 Feb 2017
Granted: 31 Dec 2019
Term ends: Not estimatedThe source records don’t support an estimate.

Classification12 codes

G01N 33/6854
Involving proteins, peptides or amino acids: immunoglobulins
C07K 16/18
Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies: against material from animals or humans
C07K 16/468
Hybrid immunoglobulins: immunoglobulins having two or more different antigen binding sites, e.g. multifunctional antibodies
G01N 21/33
Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry:…