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Studies on bioconjugation of quantum dots using capillary electrophoresis and fluorescence correlation spectroscopy
Author(s) -
Wang Jinjie,
Huang Xiangyi,
Zan Feng,
Guo Chengang,
Cao Chengxi,
Ren Jicun
Publication year - 2012
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.201200024
Subject(s) - bioconjugation , chemistry , biomolecule , capillary electrophoresis , macromolecule , conjugated system , succinimide , fluorescence , quantum dot , fluorescence spectroscopy , fluorescence correlation spectroscopy , reagent , chromatography , combinatorial chemistry , nanotechnology , organic chemistry , biochemistry , materials science , polymer , molecule , physics , quantum mechanics
In this paper, we systematically investigated the conjugation of quantum dots ( QD s) with certain biomolecules using capillary electrophoresis ( CE ) and fluorescence correlation spectroscopy ( FCS ) methods. Commercial QD s and aqueous‐synthesized QD s in our lab were used as labeling probes, certain bio‐macromolecules, such as proteins, antibodies, and enzymes, were used as mode samples, and 1‐ethyl‐3‐(3‐dimethylaminopropyl) carbodiimide hydrochloride ( EDC ) and N ‐hydroxysulfo‐succinimide ( S ulfo‐ NHS ) were used as linking reagents. We studied the effects of certain factors such as the isoelectric points (p I s) of bio‐macromolecules and buffer p H on the bioconjugation of QD s, and found that the p I s of bio‐macromolecules played an important role in the conjugation reaction. By the optimization of the buffer p H some proteins with different p I s were efficiently conjugated with QD s using EDC and S ulfo‐ NHS as linking agents. Furthermore, we on‐line investigated the kinetic process of QD s‐bioconjugation by FCS and found that the conjugation reaction of QD s with protein was rapid and the reaction process almost completed within 10 min. We also observed that QD s conjugated with proteins were stable for at least 5 days in phosphate buffer. Our work described here will be very helpful for the improvement of the QDs conjugation efficiency in bioapplications.