研究目的
Determination of factors influencing the efficiency of conjugation of various molecules with colloidal semiconductor quantum dots (QDs) is an important step toward their biomedical application.
研究成果
Electrostatic interaction governs efficiency of strain promoted [3+2] azid-alkyne cycloaddition (SPAAC) reaction for conjugation of CdSe/ZnS QDs with negatively charged molecules. Utilization of zwitter-ionic QDs with reduced surface charge allows significantly enhance the efficiency of QDs conjugation with dsDNA.
研究不足
The main disadvantage of SPAAC reaction is its lower rate compared to CuAAC, especially at low reaction concentration of the reagents. Additionally, electrostatic repulsion between negatively charged QDs and molecules can suppress the efficiency of conjugation.
1：Experimental Design and Method Selection:
Controlled strain promoted [3+2] azid-alkyne cycloaddition (SPAAC) was used to study the influence of charge interaction between QDs and molecules on the efficiency of their conjugation.
2：Sample Selection and Data Sources:
Azide-modified polymer-encapsulated core-shell CdSe/ZnS QDs, bicyclononyne(BCN)-modified JOE dye and BCN-BHQ1(Black Hole Quencher 1)-modified oligonucleotide duplex were used as model objects for conjugation.
3：List of Experimental Equipment and Materials:
CdSe/ZnS core-shell QDs, JOE dye, dsDNA-BHQ1 complex, NH2-[TEG]-N3, NH2-[TEG]-OH, EDC, MES acid, MOPS buffer, TRIS buffer, TEAA buffer, NaCl, DMSO.
4：Experimental Procedures and Operational Workflow:
Functionalization of QDs with azide groups, conjugation of QDs with JOE-BCN and dsDNA-BHQ1, monitoring of SPAAC reaction kinetics through QDs PL quenching by FRET.
5：Data Analysis Methods:
Optical absorption and PL spectroscopies, zeta potential measurements, gel filtration chromatography.
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CdSe/ZnS core-shell QDs
Model objects for conjugation
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JOE dye
Model molecule for conjugation
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dsDNA-BHQ1 complex
Model molecule for conjugation
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NH2-[TEG]-N3
Functionalization of QDs with azide groups
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NH2-[TEG]-OH
Control of azide groups number on QDs surface
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EDC
Fluka
Carbodiimide condensation
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MES acid
pH adjustment
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MOPS buffer
Buffer solution
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TRIS buffer
Buffer solution
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TEAA buffer
Buffer solution
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NaCl
Fluka
Ionic strength adjustment
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DMSO
Fluka
Solvent
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