Chromatography is a process for detaching the individual components of a mixture which according to their relative affinities for the stationary and mobile phases. The one who discover chromatography passed through a column of solid adsorbents, he coined the term chromatography, to represent the separation of color compounds. Column chromatography, the most commonly one, of which the stationary phase is put into a column through which the mobile phase is pumped. Besides, owing to its intrinsic properties, HPLC has been wildly used for the analysis of biomolecules. Based on different theories, there are different chromatographic methods being exploited. The basic procedure in HPLC is to use a column under high pressure to work with a sample which includes a liquid mobile phase, spur by a pump packed with suitable matrix. Separation is implemented through differential interaction of molecules with the column matrix and then are separated by the time required to pass the column. Then use a suitable detector to investigate signals of separated molecules and data are analyzed on the computer system with processing software. Minor differences between various proteins such as hydrophobicity, charge, bio-affinity, and size have developed some basic types of chromatography techniques, of which the approaches differ mainly in the types of stationary phase used to achieve the separation of protein molecules.
Figure 1: Fast HPLC methodAccording to the above description, it is previous that HPLC is a highly useful technique, because of its inherent characteristics of robustness, resolution, reproducibility, and high recoveries, to separate and characterize peptides and proteins. Besides, the elution of targeted protein can be easily control under different forms of HPLC techniques by altering the interactions of desired protein with stationary phase matrices. Nowadays, the tremendous growth in the field of recombinant DNA technology we have experienced has brought about rapid growth in the development of therapeutic proteins under classes of cytokines, growth factors, enzymes, hormones, blood products, and monoclonal antibodies. Thus, it is expected that HPLC in its various forms will continue to contribute importantly in the purification and analysis of peptides and proteins for a long period of time.
Creative Proteomics has a strict workflow to analyze Protein Purity Analysis by HPLC to meet your requirements.
