The OD 260/280 ratio is a valuable tool in protein purification; it serves as a guidepost for the purity and composition of a sample. This blog post will talk about the importance of measuring this value throughout the process of purifying proteins.
In 2013, crystallographer and UAB Professor Dmitry Vassylyev was purifying proteins for crystallography studies when he began to develop the CL7/Im7 system. At the time, no single affinity chromatography system existed that could give him the desired ultra-high purity, tag-free protein with reasonable yield in a short amount of time.
Biosimilars are functionally significant proteins with therapeutic potential. This blog post highlights the purification of several biosimilar proteins using the CL7/Im7 system.
Membrane proteins are difficult to purify due largely to their hydrophobic regions. When the proteins are in solution, these exposed regions can form strong non-specific interactions with each other, cytoplasmic proteins, or nucleic acids.
Multi-subunit proteins present roadblocks because of their size and the expression of multiple units. In this blog post, we discuss the one-step purification of ttRNAP and mtRNAP, two bacterial RNA polymerases, using the CL7/Im7 system.
This case study examines the Staley Lab’s success in using the CL7/Im7 system for their protein purification needs. Cody Hernandez, PhD candidate, shares his experience working with TriAltus’ methods to purify helicases, a mutant helicase, and several proteins.
Cas9 requires high purity in order to achieve high activity as a part of the CRISPR system for genetic modification. Due to a growing demand for Cas9 and an effort to improve its production, TriAltus conducted its own purification runs of Cas9 with success.
In the past several years, CRISPR has emerged as a powerful system for targeted gene modification. The system requires only two primary components: a guide RNA (gRNA or sgRNA) sequence and a Cas9 enzyme.
This blog post about pH in the context of protein purification is the third in a series about optimizing conditions for protein purification. Although pH is most commonly seen as an issue in ion-exchange chromatography, it also plays a critical role in affinity tag protein purification.
Using denaturing conditions is a way to coax insoluble proteins into solution by reducing hydrophobic effects and unfolding the aggregates. Denaturing agents are also useful for testing protein folding dynamics, protein elution from a column, and regenerating a resin column.
When conducting affinity purification of a protein, there are many variables to consider in order to optimize purity and yield. One such condition to control is high salt loading, which decreases impurities in the final product.
Protein fusion tags make affinity chromatography possible for many proteins, particularly those for which no affinity-quality antibody is available. In addition to improving conditions for purification, tags can facilitate detection and protein interaction studies.