Efficient bioconjugation of protein capture agents to biosensor surfaces using aniline-catalyzed hydrazone ligation.
Aniline-catalyzed hydrazone ligation between surface-immobilized hydrazines and aldehyde-modified antibodies is shown to be an efficient method for attaching protein capture agents to model oxide-coated biosensor substrates. Silicon photonic microring resonators are used to directly evaluate the efficiency of this surface bioconjugate reaction at various pHs and in the presence or absence of aniline as a nucleophilic catalyst. It is found that aniline significantly increases the net antibody loading for surfaces functionalized over a pH range from 4.5 to 7.4, allowing derivatization of substrates with reduced incubation time and sample consumption. This increase in antibody loading directly results in more sensitive antigen detection when functionalized microrings are employed in a label-free immunoassay. Furthermore, these experiments also reveal an interesting pH-dependent noncovalent binding trend that plays an important role in dictating the amount of antibody attached onto the substrate, highlighting the competing contributions of the bioconjugate reaction rate and the dynamic interactions that control opportunities for a solution-phase biomolecule to react with a substrate-bound reagent.
Byeon
Ji-Yeon
JY
Department of Chemistry, University of Illinois at Urbana-Champaign, 600 South Mathews Avenue, Urbana, Illinois 61801, USA.
Limpoco
F T
FT
Bailey
Ryan C
RC
D000814:Aniline Compounds / Q000737:chemistry*; D015374:Biosensing Techniques; D002384:Catalysis; D006835:Hydrazones / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D013499:Surface Properties
D000814:Aniline Compounds; D006835:Hydrazones; C023650:aniline
Anal Chem. 2010 Jan 1;82(1):69-72
20000326
Bioconjug Chem. 2007 Jan-Feb;18(1):247-53
17226979
Bioconjug Chem. 2006 Nov-Dec;17(6):1473-81
17105226
Anal Bioanal Chem. 2009 May;394(1):121-35
19221722
J Am Chem Soc. 2004 Nov 17;126(45):14730-1
15535692
Anal Chem. 2009 Nov 15;81(22):9499-506
19848413
Angew Chem Int Ed Engl. 2008;47(52):10030-74
19072788
Angew Chem Int Ed Engl. 2002 Jul 15;41(14):2596-9
12203546
Angew Chem Int Ed Engl. 2010 Jun 21;49(27):4608-11
20491114
Nat Biotechnol. 2002 Mar;20(3):270-4
11875428
J Am Chem Soc. 2006 Dec 13;128(49):15602-3
17147365
Nat Methods. 2009 Mar;6(3):207-9
19234450
J Am Chem Soc. 2010 Mar 3;132(8):2614-21
20131824
Science. 2000 Mar 17;287(5460):2007-10
10720325
Chembiochem. 2005 May;6(5):782-99
15791688
Science. 2000 Sep 8;289(5485):1760-3
10976071
Biochemistry. 1986 Sep 23;25(19):5774-9
3096375
Biosens Bioelectron. 2010 Dec 15;26(4):1283-91
20708399
Angew Chem Int Ed Engl. 2008;47(50):9618-47
19025742
Chem Rev. 2009 Sep;109(9):4025-53
19572643
Chem Rev. 2008 Feb;108(2):462-93
18229953
Angew Chem Int Ed Engl. 2006 Nov 20;45(45):7581-4
17051631
Langmuir. 2010 Aug 3;26(15):12817-23
20586451
OMICS. 2006 Fall;10(3):327-43
17069511
J Phys Chem B. 2006 Feb 2;110(4):1907-14
16471762
Bioconjug Chem. 2008 Dec;19(12):2543-8
19053314
Curr Opin Chem Biol. 2008 Dec;12(6):760-6
19058994
Langmuir. 2007 May 8;23(10):5571-7
17432882
Biomacromolecules. 2007 Jun;8(6):1775-89
17444679
Nat Chem Biol. 2005 Jun;1(1):13-21
16407987
Anal Chem. 2010 Mar 1;82(5):1975-81
20143780
J Phys Chem B. 2006 Feb 2;110(4):1553-8
16471714
Science. 1994 Nov 4;266(5186):776-9
7973629