Reversed-phase chromatography is an essential step in any drug discovery workflow, particularly as a desalting step prior to MS/MALDI analysis. One of the first choices you must make when you design your purification protocol is resin selection, and there are several options to choose from depending on your sample type and downstream application. One important variable to consider among different resin types is pore size.
Exploring the Applications of PhyTip Columns Through Many Areas in Biomolecule Drug Discovery
Since the therapeutic use of antibodies for cancer continues to grow since Kohler and Milstein’s work on the hybridoma technology, the need for high throughput antibody purification has become a staple in many areas of drug discovery and development process.
Purification of immunoglobulins is essential for the development of new biopharmaceuticals and affinity chromatography is a popular method for this application. One approach to purification of immunoglobulins it to employ histidine tags, which make binding to metal ions stronger and therefore chromatographic purification more efficient. However, when working with antibodies that are expressed without such a tag, affinity medias with immobilized ligands with binding affinity to specific regions of the antibody can be used for chromatographic purification.
High-throughput methods for protein purification and production are becoming more common in the biopharmaceutical and biotech industry. Advancements have occurred in high-throughput expression and cloning technologies. Automated protein purification methods have led to increased productivity and reproducibility. However, what technologies are available for automated high-throughput protein extraction? In the book “High-throughput protein production and purification” (link below), various authors have contributed with their protocols for their expression pipelines. This blog posts provides an overview of the automated methods for protein purification that were described here – what applications they were used for, which principles the methods are based on and how they are automated.
Immunoprecipitation is a well-known method in biomedical research for protein isolation. This method is useful to be able to study a protein’s physiochemical properties, post-translational modifications or expressions in various tissues or cell lines. In immunoprecipitation, the protein is captured on a solid support with an immobilized antibody that binds to the protein in question.
Immunoprecipitation is usually carried out in small tubes by the use of magnetic beads or loose affinity resin as solid support.
Dual flow chromatography (DFC) is the unique process of performing chromatography separations in a column bed loosely packed at the end of a pipette tip, retained by a minimally absorbing, proprietary mesh – a PhyTip®. Why? To enable automated sample purification on automated liquid handling systems while obtaining high sample recovery and purity.
For anyone looking to automate their protein expression platform, the book “High-Throughput Protein Production and Purification” that was published in July 2019 (ISBN 978-1-4939-9624-7) is an excellent read.
According to a relatively newly published journal article (Bielser et al. Aug 20 2019, J Biotech), there has been a renewed interest in the science community in perfusion cell culture technologies for continuous and efficient production of therapeutic recombinant proteins.