Advancements in Dye-Ligand Chromatography for Efficient Protein Purification: Principles, Applications and Future Prospectives
Keywords:
Affinity chromatography, Dye-ligand chromatography, Protein purification, Protein analysisAbstract
Dye-Ligand Chromatography (DLC) is a powerful and versatile technique employed in the purification of proteins. It leverages the specific interaction between a protein and a dye-functionalized ligand immobilized on a chromatographic matrix. This approach is particularly useful for separating proteins based on their affinity to the dye-ligand, providing a high degree of selectivity and efficiency. Dye-ligand chromatography has found extensive applications in various fields, including biotechnology, pharmaceuticals, and research. The primary advantage of DLC lies in its ability to purify proteins from complex mixtures, such as cell lysates, serum, and culture media, without the need for harsh chemical treatments. This technique is especially useful for isolating proteins with particular binding properties, such as enzymes, antibodies, and recombinant proteins. Furthermore, DLC can be employed for the purification of proteins that exhibit weak or transient interactions with other ligands, which are challenging to purify using traditional chromatography methods.
In addition to protein purification, dye-ligand chromatography has been applied in the analysis of protein-protein interactions, screening of enzyme inhibitors, and in the study of protein conformational changes. The ability to fine-tune the specificity of the dye-ligand system has also led to the development of novel variants for selective purification processes.
Overall, dye-ligand chromatography presents a robust and cost-effective alternative to other chromatographic methods, offering high resolution and scalability for industrial and laboratory-scale protein purification. Its adaptability and simplicity make it an indispensable tool for a wide range of protein purification applications
References
Abeyrathne EDNS, Lee HY, Ham JS, Ahn DU. Separation of ovotransferrin from chicken egg white without using organic solvents. Poultry Science. 2013; 92(4):1091–7. https://doi.org/10.3382/ps.2012-02654
Abis G, Charles RL, Eaton P, Conte MR. Expression, purification, and characterisation of human soluble Epoxide Hydrolase (Hseh) and of its functional C-terminal domain. Prot. Expr. Purif. 2018; 153:105–13. https://doi.org/10.1016/j.pep.2018.09.001
Ahmed W, Angel N, Edson J, Bibby K, Bivins A, O’Brien JW, et al. First confirmed detection of SARS-CoV-2 in untreated wastewater in Australia: A proof of concept for the wastewater surveillance of COVID-19 in the community. Sci. Total Environ. 2020; 728:138764. https://doi.org/10.1016/j.scitotenv.2020.138764
Ai Y, Davis A, Jones D, Lemeshow S, Tu H, He F, Ru P, Pan X, Bohrerova Z, Lee J. Wastewater SARS-CoV-2 monitoring as a community-level COVID-19 trend tracker and variants in Ohio, United States. Sci. Total. Environ. 2021; 801:149757. https://doi.org/10.1016/j.scitotenv.2021.149757
Akin EJ, Alsaloum M, Higerd GP, Liu S, Zhao P, Dib-Hajj FB, et al. Paclitaxel increases axonal localization and vesicular trafficking of Nav1.7. Brain. 2021; 144(6):1727–37. https://doi.org/10.1093/brain/awab113
Arora S, Saxena V, Ayyar BV. Affinity chromatography: A versatile technique for antibody purification. Methods. 2017; 116:84. https://doi.org/10.1016/j.ymeth.2016.12.010
Baghalabadi V, Doucette AA. Mass spectrometry profiling of low molecular weight proteins and peptides isolated by acetone precipitation. Analytica Chimica Acta. 2020;1138:38-48. https://doi.org/10.1016/j.aca.2020.08.057
Bhataya A, Schmidt-Dannert C, Lee PC. Metabolic engineering of Pichia pastoris X-33 for lycopene production. Process Biochemistry. 2009; 44(10):1095–102. https://doi.org/10.1016/j.procbio.2009.05.012
Bill RM, Hedfalk K. Aquaporins – Expression, purification and characterization. Biochimica et Biophysica Acta (BBA) - Bio membranes. 2021;1863(9):183650. https://doi.org/10.1016/j.bbamem.2021.183650
Bushmarina NA, Blanchet CE, Vernier G, Forge V. Cofactor effects on the protein folding reaction: Acceleration of α‐lactalbumin refolding by metal ions. Protein science. 2006;15(4):659-71. https://doi.org/10.1110/ps.051904206
Chen H, Xu Z, Xu N, Cen P. Efficient production of a soluble fusion protein containing human beta-defensin-2 in E. coli cell-free system. J. Biotechnol. 2005; 115(3):307-15. https://doi.org/10.1016/j.jbiotec.2004.08.012
Chigira Y, Oka T, Okajima T, Jigami Y. Engineering of a mammalian O-glycosylation pathway in the yeast Saccharomyces cerevisiae: production of O-fucosylated epidermal growth factor domains. Glycobiology. 2008;18(4):303–14. https://doi.org/10.1093/glycob/cwn008
Das TK, Narhi LO, Sreedhara A, Menzen T, Grapentin C, Chou DK, Antochshuk V, Filipe V. Stress factors in mAb drug substance production processes: critical assessment of impact on product quality and control strategy. J. Pharm. Sci. 2020;109(1):116-33. https://doi.org/10.1016/j.xphs.2019.09.023
Dirisala VR, Nair RR, Srirama K, Reddy PN, Rao KS, Satya Sampath Kumar N, Parvatam G. Recombinant pharmaceutical protein production in plants: unraveling the therapeutic potential of molecular pharming. Acta. Physiologiae. Plantarum. 2017; 39:1-9. https://doi.org/10.1007/s11738-016-2315-3
Fox JD. Single amino acid substitutions on the surface of Escherichia coli maltose-binding protein can have a profound impact on the solubility of fusion proteins. Protein Science. 2001; 10(3):622–30. https://doi.org/10.1110/ps.45201
