Reversed Phase HPLC Column For Protein Analysis

Reversed-phase high-performance liquid chromatography is currently the most common mode of HPLC separation, characterized by the fact that the polarity of the stationary phase is less than the polarity of the mobile phase. Protein molecules are separated due to the difference in their hydrophobicity, which allows them to partition differently in the two phases. Low pH mobile phase, room temperature or higher temperature, and the use of acetonitrile or isopropanol as the organic fraction, trifluoro as the organic fraction, and trifluoroacetic acid (TFA) as the ion-pairing agent. The separation of the proteins was more favorable. The retention index and the selectivity of the protein The selectivity was also related to the nature of the bonded stationary phase, with large pore silica gel (20-30 nm) short chain alkyl (C The short chain alkyl (C4 and C8) bonded stationary phases of large pore silica gel (20-30 nm) are suitable for protein The large pore silica gel (20~30 nm) short chain alkyl (C4 and C8) bonded stationary phases are suitable for protein separation; proteins with large molecular weight differences and very hydrophobic The separation of protein mixtures with large molecular weight differences and strong hydrophobicity was well achieved by a non-porous carrier column, and the higher column temperature also improves the separation of protein mixtures.

Fast and reliable separation of intact proteins with protein fragments

Reverse-phase techniques are used for protein identification confirmation, impurity profiling, and post-translational modification quantification. The technique is based on the difference in hydrophobicity of each substance and applies to denaturing conditions to achieve sample separation. It provides information on the amino acid sequence of the molecular level structure and sequence variants and modifications.