Conjugation of Luminescent Quantum Dots with Antibodies Using an Engineered Adaptor Protein To Provide New Reagents for Fluoroimmunoassays

Goldman, Ellen R.; Anderson, George P.; Tran, Phan T.; Mattoussi, Hedi; Charles, Paul T.; Mauro, J. Matthew

doi:10.1021/ac010662m

Cited by 407 publications

(288 citation statements)

References 22 publications

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“…Mattoussi and co-workers 25,27 were the first to use an adaptor or fusion protein for IgG antibody coupling based on electrostatic interactions. The adaptor protein has a positively charged leucine zipper domain for electrostatic binding to QDs and a protein G domain for binding to the antibody Fc region.…”

Section: Qd Bioconjugationmentioning

confidence: 99%

Bioconjugated quantum dots for multiplexed and quantitative immunohistochemistry

et al. 2007

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Bioconjugated quantum dots (QDs) provide a new class of biological labels for evaluating biomolecular signatures (biomarkers) on intact cells and tissue specimens. In particular, the use of multicolor QD probes in immunohistochemistry is considered one of the most important and clinically relevant applications. At present, however, clinical applications of QD-based immunohistochemistry have achieved only limited success. A major bottleneck is the lack of robust protocols to define the key parameters and steps. Here, we describe our recent experience, preliminary results and detailed protocols for QD-antibody conjugation, tissue specimen preparation, multicolor QD staining, image processing and biomarker quantification. The results demonstrate that bioconjugated QDs can be used for multiplexed profiling of molecular biomarkers, and ultimately for correlation with disease progression and response to therapy. In general, QD bioconjugation is completed within 1 day, and multiplexed molecular profiling takes 1-3 days depending on the number of biomarkers and QD probes used.

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Section: Qd Bioconjugationmentioning

confidence: 99%

Bioconjugated quantum dots for multiplexed and quantitative immunohistochemistry

et al. 2007

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“…These proteins show species-dependent binding affinity. [106] However, for Protein A and Protein G, the binding activities are limited by nonspecific interactions resulting from the albumin and other domains on the protein surface. [65] This finding has resulted in the development of recombinant proteins that have binding properties for a wide range of immunoglobulins and with less nonspecific interactions.…”

Section: Through Adaptor Proteins or Fc-binding Proteinsmentioning

confidence: 99%

Recent Advances in the Generation of Antibody–Nanomaterial Conjugates

Sivaram

Wardiana

Howard

et al. 2017

Adv Healthcare Materials

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Targeted nanomedicines have significantly changed the way new therapeutics are designed to treat disease. Central to successful therapeutics is the ability to control the dynamics of protein–nanomaterial interactions to enhance the therapeutic effect of the nanomedicine. The aim of this review is to illustrate the diversity and versatility of the conjugation approaches involved in the synthesis of antibody–nanoparticle conjugates, and highlight significant new advances in the field of bioconjugation. Such nanomedicines have found utility as both advanced therapeutic agents, as well as more complex imaging contrast agents that can provide both anatomical and functional information of diseased tissue. While such conjugates show significant promise as next generation targeted nanomedicines, it is recognized that there are in fact no clinically approved targeted therapeutics on the market. This fact is reflected upon within this review, and attempts are made to draw some reasoning from the complexities associated with the bioconjugation chemistry approaches that are typically utilized. Present trends, as well as future directions of next generation targeted nanomedicines are also discussed.

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“…Two key issues in utilizing QDs as fluorescent biological labels are (i) making high quality nanocrystals and (ii) choosing an appropriate surface coating technique that enables the nanocrystals to be water-soluble and functionalizable while avoiding deleterious effects on the optical properties [17,18]. To obtain robust and biocompatible QDs capable of long term stability in aqueous solution, strategies have been devised using direct adsorption of dihydrolipoic acid (DHLA) on the surface (leading to negative charges), and then electrostatic self-assembly to positively charged proteins [19,35]. Other strategies employ hydrophilic organic dendron ligands [36], and micellular encapsulation using phospholipids [17].…”

Section: Quantum Dot Bioconjugates As Fluorescence Imaging Probesmentioning

confidence: 99%

Engineering imaging probes and molecular machines for nanomedicine

Tong

Cradick

et al. 2012

Sci. China Life Sci.

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Nanomedicine is an emerging field that integrates nanotechnology, biomolecular engineering, life sciences and medicine; it is expected to produce major breakthroughs in medical diagnostics and therapeutics. Due to the size-compatibility of nano-scale structures and devices with proteins and nucleic acids, the design, synthesis and application of nanoprobes, nanocarriers and nanomachines provide unprecedented opportunities for achieving a better control of biological processes, and drastic improvements in disease detection, therapy, and prevention. Recent advances in nanomedicine include the development of functional nanoparticle based molecular imaging probes, nano-structured materials as drug/gene carriers for in vivo delivery, and engineered molecular machines for treating single-gene disorders. This review focuses on the development of molecular imaging probes and engineered nucleases for nanomedicine, including quantum dot bioconjugates, quantum dot-fluorescent protein FRET probes, molecular beacons, magnetic and gold nanoparticle based imaging contrast agents, and the design and validation of zinc finger nucleases (ZFNs) and TAL effector nucleases (TALENs) for gene targeting. The challenges in translating nanomedicine approaches to clinical applications are discussed. Nanomedicine is an emerging field that integrates nanotechnology, biomolecular engineering, biology, and medicine [1]. It focuses on the development of engineered nano-scale (1100 nm) materials, structures and devices for better diagnostics and highly specific medical intervention in curing disease or repairing damaged tissues. As a basis for nanomedicine, nanotechnology is the science, engineering, and technology related to the understanding and control of matter at the nano-scale, and the development of materials, devices, and systems that have novel properties and functions due to their nano-scale dimensions or components. Nanotechnology also provides new abilities to measure, control and manipulate matter (including soft matter) at the nano-scale what was unthinkable with conventional tools. Owing to the size-compatibility of nano-scale structures with proteins and nucleic acids in living cells, nanomedicine approaches have the potential to provide unprecedented opportunities for achieving a better control of biological processes, and drastic improvements in disease detection, therapy, and prevention, thus revolutionizing medicine. Over the last ten years or so, significant efforts have been made in the US, China, Europe and elsewhere to develop nanomedicine. For example, the US National Institutes of Health has developed a nanomedicine centers network, and invested a significant amount of research funding to nanomedicine development. Just in FY 2009 (Oct. 1, 2008Sept. 30, 2009, the total NIH funding in nanotechnology/ nanoscience projects was more than 410 million US dollars. SPECIAL TOPIC

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Conjugation of Luminescent Quantum Dots with Antibodies Using an Engineered Adaptor Protein To Provide New Reagents for Fluoroimmunoassays

Cited by 407 publications

References 22 publications

Bioconjugated quantum dots for multiplexed and quantitative immunohistochemistry

Bioconjugated quantum dots for multiplexed and quantitative immunohistochemistry

Recent Advances in the Generation of Antibody–Nanomaterial Conjugates

Engineering imaging probes and molecular machines for nanomedicine

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