The Protein Man's Blog | A Discussion of Protein Research

Proteomics is a complex field and it is exposed to major developments over the last few decades. A general proteomic experiment is a multi-step process, which involves modifications and analysis at every step depending on the experimental requirement. Crudely, these experiments have the following steps:

Maintaining protein function and stability during the entire process of protein expression and purification is crucial. Though a lot of strategies are being implemented by many researchers, each protein has its own requirements for its stability. Some proteins are prone to aggregation, a small trigger is enough to destabilise the protein at any stage of purification. Each and every step of protein purification is challenging for maintaining stability of aggregate-prone proteins. Few strategies to be considered while purifying these sensitive proteins starting from cloning to storage of pure protein.

As you are no doubt aware, carrier proteins play a significant role in antibody production. While immunogenic peptides are generally considered to be good epitopes by themselves, they are too small and lack the additional motifs required to link with class II molecules and T-cell receptors. Thus, they do not have the ability to boost the antibody response.

The main challenge during protein expression and purification is prevention of protein degradation. Whether it is an E.coli expression system or insect or mammalian, protein degradation should be minimized or avoided during purification of proteins. Protein expression at lower temperatures sometimes help in preventing the proteins from degrading. During cell lysis, proteins are exposed to harsh external conditions which result in degradation. Main reason for the protein degradation is the presence of various proteases in the cell itself.