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Characterization of the coupling of quantum dots and immunoglobulin antibodies

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Abstract

Water-soluble quantum dots (QDs) were used to label goat anti-human immunoglobulin antibodies (Abs), and the labeling process was characterized by column purification. The QDs obtained in organic solvent were modified with mercaptoacetic acid (MAA) and became water-soluble. These water-soluble QDs were linked to the antibodies using the coupling reagents ethyl-3-(dimethyl aminopropyl) carbodiimide (EDC) and N-hydroxysuccinimide (NHS). The linking process was shown to be effective by ultra-filter centrifugation and column purification. After comparing the quantities of Abs and water-soluble QDs involved in the linking reaction via column purification, it was found that a molar Abs:QD ratio of >1.2 resulted in most of the water-soluble QDs becoming covalently linked to the Abs. The circular dichroism (CD) spectra of Abs and QD–Ab conjugates were very similar to each other, indicating that the secondary structure of Abs remained largely intact after the conjugation. Finally, antigen (Ag)–antibody (Ab) recognition reactions perfomed on the surface of a glass slide showed that the conjugate retained the activity of Abs. This work lends support to the idea of linking biomolecules to QDs, and thus should aid the application of QDs to the life sciences.

Firstly in this work, the conjugates of QDs-Ab were separated from EDC&NHS in the column of Sephadex G-100(left up). Then the bioactivity of QDs-Ab was analyzed in the immunoassay (right) and the immunofluorescent signals were detected (left bottom) finally

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Acknowledgements

This work was supported by the National High Technology Research and Development Program of China (863 Program: 2003AA234010), the National Natural Science Foundation of China (Grant No.30670553, 30370387). Dr. Huang also gratefully appreciated the financial support from the National Natural Science Foundation of China (Grant No. 30400117).

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Authors and Affiliations

  1. Key Laboratory of Biomedical Photonics of Ministry of Education-Hubei Bioinformatics and Molecular Imaging Key Laboratory, College of Life Science and Technology, Huazhong University of Science and Technology, Wuhan, 430074, HuBei, People’s Republic of China

    Xiao-Feng Hua, Tian-Cai Liu, Yuan-Cheng Cao, Bo Liu, Hai-Qiao Wang, Jian-Hao Wang, Zhen-Li Huang & Yuan-Di Zhao

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  1. Xiao-Feng Hua
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  2. Tian-Cai Liu
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Correspondence to Yuan-Di Zhao.

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Hua, XF., Liu, TC., Cao, YC. et al. Characterization of the coupling of quantum dots and immunoglobulin antibodies. Anal Bioanal Chem 386, 1665–1671 (2006). https://doi.org/10.1007/s00216-006-0807-5

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  • DOI: https://doi.org/10.1007/s00216-006-0807-5

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