Research ArticleOpen AccessGoogle Scholar indexed
Rapid and Facile Purification of Apolipoprotein A-I from Human Plasma Using Thermoresponsive Nanoparticles
- 1
- 2
- 3
- 4
- 5
- 6
- 7
Journal of Biomaterials and Nanobiotechnology·Volume 02 (2011)·Pages 258–266·Published 6 July 2011·DOI10.4236/jbnb.2011.23033
Copy link · social · email
Abstract
Nanoparticles can be used to purify proteins from plasma. We report here the purification of apolipoprotein A-I (apoA-I) with high specificity from human plasma using copolymeric nanoparticles. We present an optimized protocol using 50:50 NiPAM:BAM copolymer nanoparticles with thermo-responsive properties as an affinity resin. Repeated pelleting and washing of nanoparticle-captured apoA-I is achieved through temperature cycling. The protein is then eluted using urea followed by an ion exchange step for protein concentration and depletion of nanoparticles.
KeywordsApolipoprotein A-Inanoparticlesselective purification
- H.A. Liebman, S.A. Limentani, B.C. Furie and B. Furie, “Immunoaffinity Purification of Factor-Ix (Christmas Factor) by Using Conformation-Specific Antibodies Directed against the Factor-Ix - Metal-Complex,” Proceedings of the National Academy of Sciences of the United States of America, Vol. 82, No. 11, 1985, pp. 3879-3883.
- S.O. Falkbring, P.O. G?the, P.O. Nyman, L. Sundberg and J. Porath, “Affinity Chromatography of Carbonic Anhydrase.,” FEBS Letters, Vol. 24, No. 2, 1972, pp. 229-235.
- C. Sandin, S. Linse, T. Areschoug, J.M. Woof, J. Reinholdt and G. Lindahl, “Isolation and Detection of Human IgA using a Streptococcal IgA-binding Peptide,” Journal of Immunology, Vol. 169, No. 3, 2002, pp. 1357-1364.
- K. Nord, E. Gunneriusson, J. Ringdahl, S. St?hl, M. Uhlen and P.-?. Nygren, “Binding Proteins Selected from Combinatorial Libraries of an [alpha]-Helical Bacterial Receptor Domain,” Nat Biotech, Vol. 15, No. 8, 1997, pp. 772-777.
- H.W. Gu, K.M. Xu, C.J. Xu and B. Xu, “Biofunctional Magnetic Nanoparticles for Protein Separation and Pathogen Detection,” Chemical Communications, Vol. 9, 2006, pp. 941-949.
- D.B. Shieh, C.H. Su, F.Y. Chang, Y.N. Wu, W.C. Su, J.R. Hwu, J.H. Chen, and C.S. Yeh, “Aqueous Nickel-nitrilotriacetate Modified Fe3O4-NH3+ Nanoparticles for Protein Purification and Cell Targeting,” Nanotechnology, Vol. 17, No. 16, 2006, pp. 4174-4182.
- C.C. You, A. Verma and V.M. Rotello, “Engineering the Nanoparticle-biomacromolecule Interface,” Soft Matter, Vol. 2, No. 2006, pp. 190-204.
- N.E. Miller, “Associations of High-Density-Lipoprotein Subclasses and Apolipoproteins with Ischemic-Heart- Disease and Coronary Atherosclerosis,” American Heart Journal, Vol. 113, No. 2, 1987, pp. 589-597.
- W.P. Castelli, R.D. Abbott and P.M. McNamara, “Summary Estimates of Cholesterol Used to Predict Coronary Heart-Disease,” Circulation, Vol. 67, No. 4, 1983, pp. 730-734.
- G.G. Rhoads, C.L. Gulbrandsen and A. Kagan, “Serum-Lipoproteins and Coronary Heart-Disease in a Population Study of Hawaii Japanese Men,” New England Journal of Medicine, Vol. 294, No. 6, 1976, pp. 293-298.
- C. Edelstein, D. Pfaffinger and A.M. Scanu, “Advantages and Limitations of Density Gradient Ultracentrifugation in the Fractionation of Human-Serum Lipoproteins - Role of Salts and Sucrose,” Journal of Lipid Research, Vol. 25, No. 6, 1984, pp. 630-637.
- R.J. Havel, H.A. Eder and J.H. Bragdon, “The Distribution and Chemical Composition of Ultracentrifugally Separated Lipoproteins in Human Serum,” The Journal of Clinical Investigation, Vol. 34, No. 9, 1955, pp. 1345- 1353.