Mass-transport limitations in spot-based microarrays.
Mass transport of analyte to surface-immobilized affinity reagents is the fundamental bottleneck for sensitive detection in solid-support microarrays and biosensors. Analyte depletion in the volume adjacent to the sensor causes deviation from ideal association, significantly slows down reaction kinetics, and causes inhomogeneous binding across the sensor surface. In this paper we use high-resolution molecular interferometric imaging (MI2), a label-free optical interferometry technique for direct detection of molecular films, to study the inhomogeneous distribution of intra-spot binding across 100 micron-diameter protein spots. By measuring intra-spot binding inhomogeneity, reaction kinetics can be determined accurately when combined with a numerical three-dimensional finite element model. To ensure homogeneous binding across a spot, a critical flow rate is identified in terms of the association rate k(a) and the spot diameter. The binding inhomogeneity across a spot can be used to distinguish high-affinity low-concentration specific reactions from low-affinity high-concentration non-specific binding of background proteins.
Ming
M
Zhao
Xuefeng
X
Wang
David
D
Nolte
Anal Bioanal Chem. 2005 Jan;381(1):141-55
15700161
Nat Genet. 2002 Dec;32 Suppl:526-32
12454649
Anal Biochem. 1996 Sep 5;240(2):262-72
8811920
Appl Opt. 2007 Nov 10;46(32):7836-49
17994133
Mol Cell Proteomics. 2005 Apr;4(4):377-83
15671041
Opt Express. 2006 Sep 4;14(18):8224-31
19529196
Nat Rev Drug Discov. 2002 Jul;1(7):515-28
12120258
Biosens Bioelectron. 1995;10(9-10):923-36
8652107
Biosens Bioelectron. 2008 Dec 1;24(4):987-93
18809308
Anal Chim Acta. 2008 Jul 14;620(1-2):8-26
18558119
Anal Biochem. 2007 Dec 1;371(1):105-15
17718996
Proteomics. 2006 Feb;6(3):794-803
16385475
Nat Rev Drug Discov. 2006 Apr;5(4):310-20
16582876
Biophys J. 1996 Mar;70(3):1230-49
8785280
Mol Cell Proteomics. 2006 Sep;5(9):1681-96
16735300
Proteomics Clin Appl. 2008 Oct;2(10-11):1444-59
21136793
Rev Sci Instrum. 2009 Oct;80(10):101101
19895047
Clin Chem. 1998 Sep;44(9):2015-30
9733000
Nat Mater. 2008 Jun;7(6):442-53
18497851
Expert Rev Mol Diagn. 2006 Jan;6(1):111-24
16359272
J Immunol Methods. 2004 Jul;290(1-2):107-20
15261575
Drug Discov Today. 2005 Apr 1;10(7):503-11
15809196
Expert Rev Proteomics. 2007 Aug;4(4):505-13
17705708
J Mol Recognit. 2003 Jul-Aug;16(4):165-76
12898667
Opt Express. 2008 May 12;16(10):7102-18
18545414
Clin Chem. 2006 Nov;52(11):2135-40
16990417
Nano Lett. 2005 Apr;5(4):803-7
15826132
Biosens Bioelectron. 2009 Sep 15;25(1):167-72
19628383
Biosens Bioelectron. 2007 Feb 15;22(7):1403-9
16879959
Science. 2006 Mar 17;311(5767):1592-5
16456038
J Appl Phys. 2010 Mar 15;107(6):64701
20428486
J Biol Chem. 1988 Mar 25;263(9):4323-7
2964447
Curr Opin Chem Biol. 2003 Feb;7(1):55-63
12547427
J Protein Chem. 1988 Apr;7(2):165-71
3255367