Some other advantages are that recovery of post-purification biological activity and reformation of secondary or tertiary structure after exposure to RPC are favored due to the abbreviated size of the peptides. Crude tissure extracts may be loaded directly onto the RPC system and mobilized by gradient elution. Rechromatography under the identical conditions is always an option if further purification is warranted or necessary. RPC can also be utilized in the process of protein structure determination. Extremely basic or hydrophobic proteins may not exhibit true SEC character since the columns tend to have slight hydrophobicity and anionic character.
Interaction is based on binding of the protein due to mimicry of substrate, receptor, etc.
The protein is eluted by introducing a competitive binding agent or altering the protein configuration which facilitates dissociation. RPC would be a logical choice in analysis and purification of these proteins, but IEC is also employed. This method can separate mixtures with up to 20 bases. Therefore, order of migration will start with the shorter chains and longer chains follow. Separation of tRNA can be accomplished by gradient elution where increasing organic fraction is added to the mobile phase.