Rapid and Facile Purification of Apolipoprotein A-I from Human Plasma Using Thermoresponsive Nanoparticles

Martin Lundqvist, Tord Berggård, Erik Hellstrand, Iseult Lynch, Kenneth A. Dawson, Sara Linse, Tommy Cedervall

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DOI: 10.4236/jbnb.2011.23033                

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.

Keywords

Apolipoprotein A-I, nanoparticles, selective purification

Conflicts of Interest

The authors declare no conflicts of interest.

References

[1]	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.
[2]	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.
[3]	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.
[4]	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.
[5]	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.
[6]	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.
[7]	C.C. You, A. Verma and V.M. Rotello, “Engineering the Nanoparticle-biomacromolecule Interface,” Soft Matter, Vol. 2, No. 2006, pp. 190-204.
[8]	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.
[9]	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.
[10]	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.
[11]	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.
[12]	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.
[13]	D.W. Swinkels, H.L.M. Haklemmers and P.N.M. Demacker, “Single Spin-Density Gradient Ultracentrifugation Method for the Detection and Isolation of Light and Heavy Low-Density-Lipoprotein Subfractions,” Journal of Lipid Research, Vol. 28, No. 10, 1987, pp. 1233-1239.
[14]	T. Tadey and W.C. Purdy, “Chromatographic Techniques for the Isolation and Purification of Lipoproteins,” Journal of Chromatography B: Biomedical Sciences and Applications, Vol. 671, No. 1-2, 1995, pp. 237-253.
[15]	K.H. Weisgraber, T.P. Bersot, R.W. Mahley, G. Franceschini and C.R. Sirtori, “A-I Milano Apoprotein Isolation and Characterization of a Cysteine-Containing Variant of the A-I Apoprotein from Human High Density Lipoproteins,” The Journal of Clinical Investigation, Vol. 66, No. 5, 1980, pp. 901-907.
[16]	X. Wu, R.H. Pelton, A.E. Hamielec, D.R. Woods and W. McPhee, “The Kinetics of Poly(N-Isopropylacrylamide) Microgel Latex Formation,” Colloid and Polymer Science, Vol. 272, No. 4, 1994, pp. 467-477.
[17]	I. Lynch, I. Miller, W.M. Gallagher and K.A. Dawson, “Novel Method to Prepare Morphologically Rich Polymeric Surfaces for Biomedical Applications via Phase Separation and Arrest of Microgel Particles,” Journal of Physical Chemistry B, Vol. 110, No. 30, 2006, pp. 14581-14589.
[18]	C. Oslakovic, M.J. Krisinger, A. Andersson, M. Jauhiainen, C. Ehnholm and B. Dahlback, “Anionic Phospholipids Lose Their Procoagulant Properties When Incorporated into High Density Lipoproteins,” Journal of Biological Chemistry, Vol. 284, No. 9, 2009, pp. 5896-5904.
[19]	T. Cedervall, I. Lynch, M. Foy, T. Bergg?rd, S.C. Donnelly, G. Cagney, S. Linse, and K.A. Dawson, “Detailed Identification of Plasma Proteins Adsorbed on Copolymer Nanoparticles,” Angewandte Chemie International Edition, Vol. 46, No. 30, 2007, pp. 5754-5756.
[20]	T. Cedervall, I. Lynch, S. Lindman, T. Berggard, E. Thulin, H. Nilsson, K.A. Dawson, and S. Linse, “Understanding the Nanoparticle-protein Corona using Methods to Quantify Exchange Rates and Affinities of Proteins for Nanoparticles,” Proceedings of the National Academy of Sciences of the United States of America, Vol. 104, No. 7, 2007, pp. 2050-2055.
[21]	M. Lundqvist, I. Sethson and B.H. Jonsson, “Protein Adsorption onto Silica Nanoparticles: Conformational Changes Depend on the Particles’ Curvature and Protein Stability,” Langmuir, Vol. 20, No. 2004, pp. 10639- 10647.