Use Of Matrix-Based Experimental Design For Protein Stability Using Design Of Experiment And High Throughput Screening: Evaluation Of The Stability Of Human Serum Albumin Using An ICH Verified Reversed Phase High Performance Liquid Chromatography Method

Location

SU 215

Start Date

19-4-2019 10:00 AM

Department

Chemistry

Abstract

In biopharmaceutical product development and subsequent characterization, there are two classes of impurities that can be harmful to humans: process related impurities and product related impurities. Process related impurities are expressed as a by-product of bacterial, mammalian, viral, plasma or plant expression systems such as host cell proteins, host DNA, endotoxins, endogenous virus, or simply bacterial contamination. These by-products can compromise the safety, quality, and efficacy of a biological drug product. With these impurities, there are readily known process steps to remove, such as membrane/media based anion-exchange chromatography, hydrophobic interaction chromatography, and viral/sterile filtration. The other major classes of impurities for biologically related products are known as product related impurities, which can result from post-translational modification to the target molecule. These are product related impurities that are the same as the molecule of interest, but degraded in a certain way such as oxidation, deamidation, aggregation, or reduction of disulfide bonds when present. More than one factor at a time such as temperature, additive concentration, or pH can play a role in generation of such product related impurities. This work examines the use of design of experiment (DoE) and high throughput screening (HTS) after the development and verification of a reversed phase high performance chromatography method to examine of multivariate parameters impact on native, monomeric human serum albumin protein stability as a preliminary step in the development of a RP-HPLC peptide map post cyanogen bromide digestion.

Comments

John Al-Bazi is the faculty sponsor of this project.

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Apr 19th, 10:00 AM

Use Of Matrix-Based Experimental Design For Protein Stability Using Design Of Experiment And High Throughput Screening: Evaluation Of The Stability Of Human Serum Albumin Using An ICH Verified Reversed Phase High Performance Liquid Chromatography Method

SU 215

In biopharmaceutical product development and subsequent characterization, there are two classes of impurities that can be harmful to humans: process related impurities and product related impurities. Process related impurities are expressed as a by-product of bacterial, mammalian, viral, plasma or plant expression systems such as host cell proteins, host DNA, endotoxins, endogenous virus, or simply bacterial contamination. These by-products can compromise the safety, quality, and efficacy of a biological drug product. With these impurities, there are readily known process steps to remove, such as membrane/media based anion-exchange chromatography, hydrophobic interaction chromatography, and viral/sterile filtration. The other major classes of impurities for biologically related products are known as product related impurities, which can result from post-translational modification to the target molecule. These are product related impurities that are the same as the molecule of interest, but degraded in a certain way such as oxidation, deamidation, aggregation, or reduction of disulfide bonds when present. More than one factor at a time such as temperature, additive concentration, or pH can play a role in generation of such product related impurities. This work examines the use of design of experiment (DoE) and high throughput screening (HTS) after the development and verification of a reversed phase high performance chromatography method to examine of multivariate parameters impact on native, monomeric human serum albumin protein stability as a preliminary step in the development of a RP-HPLC peptide map post cyanogen bromide digestion.