ACRYLAMIDE PHOTOPOLYMERIZATION FOR ANALYTICAL ELECTROPHORESIS
DOI:
https://doi.org/10.32471/oncology.2663-7928.t-24-3-2022-g.10692Keywords:
biomarkers, electrophoresis, photopolymerization, protein separationAbstract
Aim: To create a low-toxicity catalytic system for the photopolymerization of polyacrylamide gels. To study the suitability of the proposed system for electrophoretic separation of blood proteins. Object and methods: The samples of donors peripheral blood has been studed. Photopolymerization of acrylamide/N,N′-methylenebisacrylamide was assisted with combination of ethylenediaminetetraacetic acid disodium dihydrate catalyst, riboflavin photoinitiator, and artificial daylight (wavelength range of λ=400–700 nm). Results: a new catalytic system for photopolymerization of polyacrylamide gels was created. The suitability of the proposed method for electrophoretic separation of blood proteins is shown. Conclusions: the use of the proposed method can be particularly appropriate for the study of molecular heterogeneity of enzymes, which can provide additional data on the course and prognosis of cancer.
References
Magdeldin S. Gel electrophoresis: Principles and basics. Rijeka, Croatia: InTech, 2012. doi:10.5772/2205
Kurien BT, Scofield RH. Electrophoretic Separation of Proteins. New York: Springer, 2019. https://link.springer.com/book/10.1007/978-1-4939-8793-1.
Westermeier R. Electrophoresis in practice: a guide to methods and applications of DNA and protein separations. Freiburg, Germany: John Wiley & Sons, 2016. 406p.
Lledias F, Hernбndez F, Rivas V, et al. A rapid and reliable method for total protein extraction from succulent plants for proteomic analysis. Protein J 2017; 36 (4): 308–321. 10.1007/s10930-017-9739-5.
Iwashita K, Inoue N, Handa A, et al. Thermal aggregation of hen egg white proteins in the presence of salts. Protein J 2015; 34 (3): 212–219. doi.org/10.1007/s10930-015-9612-3
Sharma N, Sharma R, Rajput YS, et al. Separation methods for milk proteins on polyacrylamide gel electrophoresis: Critical analysis and options for better resolution. Int Dairy J 2020; 104920. doi:10.1016/j.idairyj.2020.104920
Corbo C, Cevenini A, Salvatore F. Biomarker discovery by proteomics-based approaches for early detection and personalized medicine in colorectal cancer. Proteomics — Clin Appl 2017; 11 (5-6): 1600072. doi:10.1002/prca.201600072
Sobsey CA, Ibrahim S, Richard VR, et al. Targeted and untargeted proteomics approaches in biomarker development. Proteomics 2020; 20 (9): 1900029. doi:10.1002/pmic.201900029
Kapitany RA, Zebrowski EJ. A high resolution PAS stain for polyacrylamide gel electrophoresis. Anal Biochem 1973; 56 (2): 361–369. doi.org/10.1016/0003-2697(73)90202-9
Brunelle JL, Green R. Coomassie blue staining. In: Methods in enzymology. Elsevier, 2014: 161–167. doi:10.1016/B978-0-12-420119-4.00013-6
Krause RGE, Goldring JPD. Crystal violet stains proteins in SDS-PAGE gels and zymograms. Anal Biochem 2019; 566: 107–115. doi:10.1016/j.ab.2018.11.015
Sennakesavan G, Mostakhdemin M, Dkhar LK, et al. Acrylic acid/acrylamide based hydrogels and its properties — A review. Polym Degrad Stab 2020; 180: 109308. doi:10.1016/j.polymdegradstab.2020.109308
Righetti PG, Sebastiano R, Citterio A. Capillary electrophoresis and isoelectric focusing in peptide and protein analysis. Proteomics 2013; 13 (2): 325–340. doi:10.1002/pmic.201200378
Davis BJ. Disc electrophoresis. II. Method and application to human serum proteins. Ann NY Acad Sci 1964; 121 (2): 404–427.
Rabilloud T, Vincon M, Garin J. Micropreparative one-and two-dimensional electrophoresis: Improvement with new photopolymerization systems. Electrophoresis 1995; 16 (1): 1414–1422. doi:10.1002/elps.11501601234
Sheth S, Jain E, Karadaghy A, et al. UV dose governs UV-polymerized polyacrylamide hydrogel modulus. Int J Polym Sci 2017; 2017: ID 5147482. doi:10.1155/2017/5147482
Mane S, Ponrathnam S, Chavan N. Effect of chemical cross-linking on properties of polymer microbeads: A review. Can Chem Trans 2015; 3 (4): 473–485. doi:10.13179/canchemtrans.2015.03.04.0245
Zarei M, Zarei M, Ghasemabadi M. Nanoparticle improved separations: From capillary to slab gel electrophoresis. TrAC Trends Anal Chem 2017; 86: 56–74. doi:10.1016/j.trac.2016.11.004
Green MR, Sambrook J. Polyacrylamide gel electrophoresis. Cold Spring Harb Protoc 2020; 2020 (12): 525–532. doi:10.1101/pdb.prot100412
Allison JH, Agrawal HC, Moore BW. Effect of N, N, N′, N′-tetramethylethylenediamine on the migration of proteins in SDS polyacrylamide gels. Anal Biochem 1974; 58 (2): 592–601. doi.org/10.1016/0003-2697(74)90228-0
Lyubimova T, Caglio S, Gelfi C, et al. Photopolymerization of polyacrylamide gels with methylene blue. Electrophoresis 1993; 14 (1): 40–50. doi:10.1002/elps.1150140108
Papavassiliou AG. Footprinting DNA-protein interactions in native polyacrylamide gels by chemical nucleolytic activity of 1, 10-phenanthroline-copper. Methods Mol Biol 2009; 543: 163–99. doi: 10.1007/978-1-60327-015-1_13.
Carbonara K, Coorssen JR. A ‘green’ approach to fixing polyacrylamide gels. Anal Biochem 2020; 605: 113853. doi:10.1016/j.ab.2020.113853
Lanigan RS, Yamarik TA. Final report on the safety assessment of EDTA, calcium disodium EDTA, diammonium EDTA, dipotassium EDTA, disodium EDTA, TEA-EDTA, tetrasodium EDTA, tripotassium EDTA, trisodium EDTA, HEDTA, and trisodium HEDTA. Int J Toxicol 2002; 21: 95–142. doi: 10.1080/10915810290096522
Ferrero ME. Rationale for the successful management of EDTA chelation therapy in human burden by toxic metals. Biomed Res Int 2016; 2016: ID 8274504. doi:10.1155/2016/8274504
Suwannasom N, Kao I, Pruβ A, et al. Riboflavin: The Health Benefits of a Forgotten Natural Vitamin. Int J Mol Sci 2020; 21 (3): 950. doi:10.3390/ijms21030950
Kim E, Kim MH, Song JH, et al. Dual crosslinked alginate hydrogels by riboflavin as photoinitiator. Int J Biol Macromol 2020; 154: 989–98. doi:10.1016/j.ijbiomac.2020.03.134
Sheraz MA, Kazi SH, Ahmed S, et al. Photo, thermal and chemical degradation of riboflavin. Beilstein J Org Chem 2014; 10: 1999–2012. doi:10.3762/bjoc.10.208
Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 1970; 227: 680–685. doi:10.1038/227680a0
Aoki T. A comprehensive review of our current understanding of red blood cell (RBC) glycoproteins. Membranes (Basel) 2017; 7 (4): 56. doi:10.3390/membranes7040056
Ryu KE, Rhim H, Park CW, et al. Plasma protein adsorption to anion substituted poly (vinyl alcohol) membranes. Macromol Res 2003; 11 (6): 451–457. doi.org/10.1007/BF03218975
Lavrentieva A. Gradient Hydrogels. Adv Biochem Eng Biotechnol. 2021;178:227-251. doi: 10.1007/10_2020_155.
Majcher MJ, Hoare T. Applications of Hydrogels 14. In: Mazumder MAJ, Sheardown H, Al-Ahmed A, editors. Functional biopolymers. Springer Nature Switzerland, 2019: 453–90. doi:10.1007/978-3-319-95990-0_17
Vandekerckhove J, Bauw G, Van Den Bulcke M, et al. Laboratory Methodology in Biochemistry. In: Fini C, Floridi A, Finelli VN, Wittman-Liebold B, editors. CRC Press, 2019: 217–238. doi:10.1201/9781351073929
Schägger H. Tricine–SDS-PAGE. Nat Protoc 2006; 1 (1): 16–22. doi.org/10.1038/nprot.2006.4
