Applications of FPLC:
Advantages of Fast Liquid Protein Chromatography:
Drawbacks:
- Ion Exchange Chromatography:
- Hydroxyapatite chromatography:
- Affinity Chromatography:
- Reverse-Flow chromatography for Affinity Purifications:
- Gel Filtration Chromatography:
- Hydrophobic Interaction Chromatography:
Advantages of Fast Liquid Protein Chromatography:
- FPLC is the fast and simple chromatography system for the separation of various proteins present in body fluids such as plasma, urine and specially cerebrospinal fluid (CSF).
- It is used for the separation of peptides, polynucleotides.
- Used for the purification of synthetic oligonucleotides.
- Used for the purification of plasmid DNA.
- Rapid purification of RNA.
- Used for the identification of protein profiles and variability within single proteins.
- It can separate the proteins from a more complex mixture in the liquid phase.
- Used in the enzymology and biology.
- Lipoprotein can be separated by FPLC.
- For the analysis of nitrogenous constituents of beer.
- Used to measure the levels of Tubular Protenuria: During Tubular Protenuria, patient’s kidney secrets more than 150mg of urinary proteins daily. By using FPLC, the specific proteins can be isolated from the extracted proteins.
- Profiling of plasma proteins in cerebrospinal fluid: Proteins in cerebrospinal fluid or CSF can be profiled by using anion exchange column in FPLC. FPLC can easily track changes occurring in patient’s body.
- Analysing Pancreatitis: In pancreatic juice, certain type of proteins can be determined by using FPLC, which lead to disease.
- By FPLC peptide sequencing and protein structures/functions can be studied.
Drawbacks:
- Needs glass columns.
- FPLC cannot handle high pressure.
- Instrument does not support HPLC columns.
- A purifying thermo labile (heat sensitive) protein is a hard task.