Site-selective in situ growth of fluorescent polymer–antibody conjugates with enhanced antigen detection by signal amplification

Zhao, Wenxiang; Liu, Xinyu; Wang, Guilin; Wang, Yang; Liu, Dong; Xie, Liangzhi; Gao, Yan; Deng, Haiteng; Gao, Wanzhen

doi:10.1016/j.biomaterials.2015.06.020

Cited by 27 publications

(28 citation statements)

References 30 publications

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“…To further improve on the design, Gao et al. have recently used DSMs to install a macroinitiator into interchain disulfide in antibodies for the in situ atom‐transfer radical‐polymerization (ATRP) of dye or drug‐labelled polymer conjugates to introduce high payload and greater possibility to fine‐tune dye‐to‐antibody ratios in a homogenous polymer–antibody hybrid . The antibody hybrid reported in this proof‐of‐concept work shows enhanced antigen detection and is far superior to a conventional antibody–dye conjugate in antigen detection by signal amplification …”

Section: Preparation Of Functional Biohybrids and Application Higmentioning

confidence: 98%

“…A fluorescent tag was also incorporated site‐selectively into lysozyme, an enzyme which hydrolyses the β (l–4) glycosidic bonds, and the mono‐modified enzyme retains 89 % activity . On the other hand, DSMs and dibromopyridazinediones have been employed extensively in the modification of peptides and antibodies to introduce bioorthogonal handles, macroinitiators, imaging probes, spin labels and drugs …”

Section: Disulfide Rebridging Agents: Syntheses Features and Biocmentioning

confidence: 99%

“…[34,35] A fluorescent tag was also incorporated site-selectively into lysozyme, an enzyme which hydrolyses the b(l-4) glycosidic bonds, and the mono-modified enzyme retains 89 % activity. [60] On the other hand, DSMs and dibromopyridazinediones have been employed extensively in the modification of peptides and antibodies to introduce bioorthogonal handles, [55] macroinitiators, [62] imaging probes, [43] spin labels [63] and drugs. [46,49] Multimodal disulfide rebridging reagents Besides, mono-functionalization, the development of multimodal reagents for increasing the functionality of proteins has received emerging interest over the past years.…”

Section: Synthetic Toolbox For Functionalizationmentioning

confidence: 99%

“…[62] The antibody hybrid reported in this proof-of-concept work shows enhanced antigen detection and is far superior to a conventional antibody-dye conjugate in antigen detection by signal amplification. [62] On the other hand, the preparation of "bispecifics" that comprise two antibody fragments that can simultaneously bind two different antigens has been introduced to improve the therapeutic efficacy. [113] In this case, Fab and disulfide-stabilized single-chain variable antibody fragments (scFv), which are commonly applied in a range of bispecific topologies, are chosen as the two components.…”

Section: Antibody Biohybridsmentioning

confidence: 99%

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Site‐Selective Disulfide Modification of Proteins: Expanding Diversity beyond the Proteome

Kuan

Wang

Weil

2016

Chemistry A European J

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The synthetic transformation of polypeptides with molecular accuracy holds great promise for providing functional and structural diversity beyond the proteome. Consequently, the last decade has seen an exponential growth of site‐directed chemistry to install additional features into peptides and proteins even inside living cells. The disulfide rebridging strategy has emerged as a powerful tool for site‐selective modifications since most proteins contain disulfide bonds. In this Review, we present the chemical design, advantages and limitations of the disulfide rebridging reagents, while summarizing their relevance for synthetic customization of functional protein bioconjugates, as well as the resultant impact and advancement for biomedical applications.

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Section: Preparation Of Functional Biohybrids and Application Higmentioning

confidence: 98%

Section: Disulfide Rebridging Agents: Syntheses Features and Biocmentioning

confidence: 99%

Section: Synthetic Toolbox For Functionalizationmentioning

confidence: 99%

Section: Antibody Biohybridsmentioning

confidence: 99%

See 2 more Smart Citations

Site‐Selective Disulfide Modification of Proteins: Expanding Diversity beyond the Proteome

Kuan

Wang

Weil

2016

Chemistry A European J

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“…Inspired by this work, we chose to synthesize C-terminal IFN-α conjugates of POEGMA with three high molecular weights of ca. 20, 60 and 100 kDa by using the SIG method that has recently been developed by us [18][19][20][21][22]. We found that increasing the molecular weight of POEGMA decreased the in vitro activity of IFN-α but increased the in vitro stability of IFN-α.…”

mentioning

confidence: 88%

Tuning the molecular size of site-specific interferon-polymer conjugate for optimized antitumor efficacy

Wang

Gao

2017

Sci. China Mater.

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The covalent attachment of protein-resistant polymers to therapeutic proteins is a widely used method for extending their in vivo half-lives; however, the effect of molecular weight of polymer on the in vitro and in vivo functions of protein-polymer conjugates has not been well elucidated. Herein we report the effect of molecular weight of poly(oligo(ethylene glycol) methyl ether methacrylate) (POEGMA) on the in vitro and in vivo properties of C-terminal interferon-alpha (IFN)-POEGMA conjugates. Increasing the molecular weight of POEGMA decreased the in vitro activity of IFN-α but increased its thermal stability and in vivo pharmacokinetics. Intriguingly, the in vivo antitumor efficacy of IFN-α was increased by increasing the POEGMA molecular weight from ca. 20 to 60 kDa, but was not further increased by increasing the molecular weight of POEGMA from ca. 60 to 100 kDa due to the neutralization of the improved pharmacokinetics and the reduced in vitro activity. This finding offers a new viewpoint on the molecular size rationale for designing next-generation protein-polymer conjugates, which may benefit patients by reducing administration frequency and adverse reactions, and improving therapeutic efficacy.

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DNA‐Mediated, On‐Membrane Sequential Assembly of Conjugated Polymer Nanoparticles for Sensitive Detection of Cell Surface Markers

Maeda,

Nakamura,

Ohta

2024

Adv Funct Materials

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Flow cytometry can provide detailed information about protein expression on cell surface and is, therefore, widely used in clinical testing. However, owing to the limited sensitivity of fluorescence signals, detection of low‐expression cell surface markers is challenging. The present report describes a DNA‐mediated, on‐membrane assembly of conjugated polymer nanoparticles (Pdots) that amplifies fluorescence signal from surface markers for sensitive detection via flow cytometry. Single‐stranded DNA (ssDNA)‐conjugated antibodies are first bound to cell surface markers, from which ssDNA‐modified Pdots are sequentially assembled using DNA hybridization. The use of DNA as a linker enables the distance‐controlled assembly of Pdots to prevent fluorescence quenching, whereas their on‐membrane sequential assembly allows amplification of the fluorescence signal without reducing binding ability of antibodies. Thus, two rounds of Pdot assembly achieve 31‐fold amplification of the fluorescence signal from CD19 on Nalm‐6 cells, which is 125‐fold brighter than that obtained using the conventional fluorescent dye‐based method. Moreover, the sequential assembly of 22 nm Pdots shows 24‐fold higher fluorescence than one‐step labeling with 81 nm Pdots, suggesting the advantage of the sequential assembly strategy in avoiding steric hindrance. The proposed method is expected to contribute to the sensitive detection of low‐expression surface markers for early and accurate diagnosis.

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Site-selective in situ growth of fluorescent polymer–antibody conjugates with enhanced antigen detection by signal amplification

Cited by 27 publications

References 30 publications

Site‐Selective Disulfide Modification of Proteins: Expanding Diversity beyond the Proteome

Site‐Selective Disulfide Modification of Proteins: Expanding Diversity beyond the Proteome

Tuning the molecular size of site-specific interferon-polymer conjugate for optimized antitumor efficacy

DNA‐Mediated, On‐Membrane Sequential Assembly of Conjugated Polymer Nanoparticles for Sensitive Detection of Cell Surface Markers

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