Regeneron Pharmaceuticals, Inc.

Hongxia Wang

Ning Li

Haibo Qiu

Jikang Wu

Xueqing Zhao

Sarah Nicoletti

Victor Shashilov

Abdelqader Zamamiri

Reginald Smith

Disclosed are methods for improving compound detection and characterization. Methods for characterizing a sample are disclosed. The methods can include providing a sample to a liquid chromatography system capable of sample separation to generate sample components; analyzing sample components by multiplexed targeted selected ion monitoring (SIM) to generate an inclusion list; and performing iterative mass spectral data-dependent acquisition (DDA) from the inclusion list, to identify individual sample components thereby characterizing the sample. In one example, multiplexed targeted SIMs and iterative MS2 DDA acquisition is used to increase robust compound identification for cell culture medium analysis.

High confidence compound identification by liquid chromatography-mass spectrometry

The present invention provides methods to quickly and efficiently assess the effect of antibody variants, including PTMs, e.g., glycosylation, oxidation, and deamidation variants, on the binding between the antibody and FcRn. In particular, the present invention is based, at least in part, on the development of an online, two dimension liquid chromatography (2D-LC) method for assessing binding between antibody variants and FcRn. In one aspect, the online 2D-LC is coupled with mass spectrometry (MS). The present invention allows for differentiation of antibody variants by peak pattern and retention time profile by affinity chromatography, and identification of these variants by mass spectrometry analysis.

Methods for assessing binding affinity of an antibody variant to the neonatal Fc receptor

Methods for detecting and/or discriminating between post-translational modification variants of an antibody of interest in a sample. The methods including: contacting a sample comprising one or more antibodies of interest with a protease to digest the sample into antibody fragments; separating antibody fragments by molecular weight and/or charge in one or more capillaries using capillary electrophoresis; eluting separated antibody fragments from the one or more capillaries; and determining the mass of the eluted antibody fragments by mass spec analysis, thereby detecting and/or discriminating between post-translational modification variants of the antibody of interest.

Native microfluidic CE-MS analysis of antibody charge heterogeneity

Methods of improving recombinant protein titer and cell titer in cell culture using cell culture media having reduced impurities are provided, and well as cell culture media having reduced impurities that can used for the production of a recombinant protein and cells with improved titer. The cell culture media having reduced impurities comprises a HEPES buffer, and the reduced impurities are HEPES related impurities. In certain aspects, methods and media improve protein titer, cell growth, and/or viable cell density.

Growing community of inventors

Briarcliff Manor, NY, United States of America

Hongxia Wang