Synthesis of Protein-Oligonucleotide Conjugates

Watson, Emma E.; Winssinger, Nicolas

doi:10.3390/biom12101523

Cited by 2 publications

(2 citation statements)

References 56 publications

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“…Nonetheless, the potential of protein microarrays, as opposed to DNA arrays, is still constrained by a number of technological obstacles. While DNA is a relatively stable oligomer with well-defined physicochemical characteristics, practically independent of its sequence, proteins refer to a heterogeneous group of biomolecules that are generally more fragile than DNA [ 4 ]. As a result, selecting an optimal immobilization approach that assures protein availability and precise surface orientation is critical.…”

Section: Introductionmentioning

confidence: 99%

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A Careful Insight into DDI-Type Receptor Layers on the Way to Improvement of Click-Biology-Based Immunosensors

Karoń,

Drozd,

Malinowska

2024

Biosensors

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Protein-based microarrays are important tools for high-throughput medical diagnostics, offering versatile platforms for multiplex immunodetection. However, challenges arise in protein microarrays due to the heterogeneous nature of proteins and, thus, differences in their immobilization conditions. This article advocates DNA-directed immobilization (DDI) as a solution, emphasizing its rapid and cost-effective fabrication of biosensing platforms. Thiolated single-stranded DNA and its analogues, such as ZNA® and PNA probes, were used to immobilize model proteins (anti-CRP antibodies and SARS-CoV nucleoprotein). The study explores factors influencing DDI-based immunosensor performance, including the purity of protein-DNA conjugates and the stability of their duplexes with DNA and analogues. It also provides insight into backfilling agent type and probe surface density. The research reveals that single-component monolayers lack protection against protein adsorption, while mixing the probes with long-chain ligands may hinder DNA-protein conjugate anchoring. Conventional DNA probes offer slightly higher surface density, while ZNA® probes exhibit better binding efficiency. Despite no enhanced stability in different ionic strength media, the cost-effectiveness of DNA probes led to their preference. The findings contribute to advancing microarray technology, paving the way for new generations of DDI-based multiplex platforms for rapid and robust diagnostics.

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Section: Introductionmentioning

confidence: 99%

“…Conjugating the two biomolecules is also quite troublesome to control, regardless of the strategy employed. Nevertheless, conjugate valency (DNA to protein ratio) aspects are also underestimated in constructing DDI-based bioreceptor platforms [ 4 , 18 ].…”

Section: Introductionmentioning

confidence: 99%

A Careful Insight into DDI-Type Receptor Layers on the Way to Improvement of Click-Biology-Based Immunosensors

Karoń,

Drozd,

Malinowska

2024

Biosensors

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Enhancing the Effectiveness of Oligonucleotide Therapeutics Using Cell-Penetrating Peptide Conjugation, Chemical Modification, and Carrier-Based Delivery Strategies

2023

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Oligonucleotide-based therapies are a promising approach for treating a wide range of hard-to-treat diseases, particularly genetic and rare diseases. These therapies involve the use of short synthetic sequences of DNA or RNA that can modulate gene expression or inhibit proteins through various mechanisms. Despite the potential of these therapies, a significant barrier to their widespread use is the difficulty in ensuring their uptake by target cells/tissues. Strategies to overcome this challenge include cell-penetrating peptide conjugation, chemical modification, nanoparticle formulation, and the use of endogenous vesicles, spherical nucleic acids, and smart material-based delivery vehicles. This article provides an overview of these strategies and their potential for the efficient delivery of oligonucleotide drugs, as well as the safety and toxicity considerations, regulatory requirements, and challenges in translating these therapies from the laboratory to the clinic.

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Synthesis of Protein-Oligonucleotide Conjugates

Cited by 2 publications

References 56 publications

A Careful Insight into DDI-Type Receptor Layers on the Way to Improvement of Click-Biology-Based Immunosensors

A Careful Insight into DDI-Type Receptor Layers on the Way to Improvement of Click-Biology-Based Immunosensors

Enhancing the Effectiveness of Oligonucleotide Therapeutics Using Cell-Penetrating Peptide Conjugation, Chemical Modification, and Carrier-Based Delivery Strategies

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