Self-interaction chromatography of proteins on a microfluidic monolith.
A novel miniaturized system has been developed for measuring protein-protein interactions in solution with high efficiency and speed, and minimal use of protein. A chromatographic monolith synthesized in a capillary is used in the method to make interaction measurements by self-interaction chromatography (SIC) in a manner that, compared to column methods, is more efficient as well as more readily practicable even if only small amounts of protein are available. The microfluidic monolith requires much less protein for both column synthesis and the chromatographic measurements than a conventional SIC system, and in addition offers improved mass transfer and hence higher chromatographic efficiency than for previous SIC miniaturization systems. Protein self-interactions for catalase as a model protein, quantified by measurement of second virial coefficients, B(22), were determined by SIC and follow trends that are consistent with previously reported values. Different column derivatization conditions were studied in order to optimize the chromatographic behavior of the microfluidic system for SIC measurements. Chromatographic sensitivity can be further increased by using different column synthesis conditions.
Department of Chemical Engineering, University of Delaware, Newark, DE 19716, USA.
Cristina
C
Martin
Abraham M
AM
Lenhoff
Biochim Biophys Acta. 2008 Apr;1784(4):600-10
18258214
Biophys J. 2005 May;88(5):3300-9
15792969
Biophys J. 2008 Jan 15;94(2):570-83
18160663
Biophys J. 2007 Jan 1;92(1):234-44
17028129
J Chromatogr A. 2005 Sep 30;1089(1-2):111-24
16130779
Anal Chem. 2003 Oct 15;75(20):5328-35
14710809
Biotechnol Prog. 2003 Mar-Apr;19(2):575-9
12675603
Acta Crystallogr D Biol Crystallogr. 2002 Mar;58(Pt 3):472-9
11856833
J Chromatogr A. 2000 Aug 4;888(1-2):1-12
10949467
Proc Natl Acad Sci U S A. 1955 Oct 15;41(10):710-4
16589733
Acta Crystallogr D Biol Crystallogr. 2002 Oct;58(Pt 10 Pt 1):1531-5
12351855
Protein Sci. 2007 Sep;16(9):1867-77
17766383
Acta Crystallogr D Biol Crystallogr. 1994 Jul 1;50(Pt 4):361-5
15299385
Curr Opin Biotechnol. 2003 Oct;14(5):512-6
14580581
Anal Biochem. 2007 Dec 1;371(1):21-5
17723222
Biophys J. 2005 Dec;89(6):4211-8
16199499
Biophys J. 2002 Mar;82(3):1620-31
11867474
J Chromatogr A. 2000 Jun 23;883(1-2):39-54
10910199
Biopolymers. 2006;84(5):527-33
16767741
Biotechnol Prog. 2003 May-Jun;19(3):1006-10
12790668
Acta Crystallogr D Biol Crystallogr. 2005 Jun;61(Pt 6):724-30
15930629
Eur Biophys J. 1998;27(3):263-71
9615398
J Chromatogr A. 2004 May 28;1037(1-2):273-82
15214670
Biophys J. 1998 Dec;75(6):2682-97
9826592
Anal Biochem. 2006 Oct 1;357(1):35-42
16843425
Proteins. 2003 Feb 1;50(2):303-11
12486724
J Sep Sci. 2004 Jul;27(10-11):747-66
15354553
Acta Crystallogr D Biol Crystallogr. 2010 Jan;66(Pt 1):44-50
20057048
Lab Chip. 2009 Feb 21;9(4):600-5
19190796
Anal Chem. 2002 Aug 15;74(16):4081-8
12199578
Anal Chem. 1992 Jun 1;64(11):1232-8
1621992
J Chromatogr A. 2007 Jan 26;1140(1-2):140-6
17182044
Biotechnol Bioeng. 1996 Oct 20;52(2):193-203
18629888