scholarly journals Carbohydrate-Encapsulated Gold Nanoparticles for Rapid Target-Protein Identification and Binding-Epitope Mapping

ChemBioChem ◽  
2005 ◽  
Vol 6 (7) ◽  
pp. 1169-1173 ◽  
Author(s):  
Yu-Ju Chen ◽  
Shu-Hua Chen ◽  
Yuh-Yih Chien ◽  
Yu-Wan Chang ◽  
Hsin-Kai Liao ◽  
...  
Keyword(s):  
Gold Nanoparticles ◽  
Epitope Mapping ◽  
Protein Identification ◽  
Target Protein ◽  
Binding Epitope

Poly(N-isopropylacrylamide)-Stabilized Gold Nanoparticles in Combination with Tricationic Branched Phenylene-Ethynylene Fluorophore for Protein Identification

Langmuir ◽  
2013 ◽  
Vol 29 (39) ◽  
pp. 12317-12327 ◽  
Author(s):  
Keerati Kusolkamabot ◽  
Pornpen Sae-ung ◽  
Nakorn Niamnont ◽  
Kanet Wongravee ◽  
Mongkol Sukwattanasinitt ◽  
...  
Keyword(s):  
Gold Nanoparticles ◽  
Protein Identification ◽  
Phenylene Ethynylene

The binding epitope of sintilimab on PD-1 revealed by AbMap

2021 ◽  
Author(s):  
Mingliang Ma ◽  
Huan Qi ◽  
Chuansheng Hu ◽  
Zhaowei Xu ◽  
Fanlin Wu ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Structural Analysis ◽  
Western Blot ◽  
Epitope Mapping ◽  
High Efficiency ◽  
Point Mutations ◽  
Therapeutic Antibodies ◽  
Binding Interface ◽  
Binding Epitope ◽  
Key Residues

Abstract PD-1 plays an important role as an immune checkpoint. Sintilimab is a newly approved PD-1 antibody for cancer immunotherapy with an unknown binding epitope on PD-1. In this study, to elucidate the molecular mechanism by which sintilimab blocks PD-1 activation, we applied Antibody binding epitope Mapping (AbMap) to identify the binding epitope of sintilimab. An epitope was successfully identified, i.e. SLAPKA, aa 127–132. By constructing a series of point mutations, the dominant residues S127, L128, A129, P130, and A132 of PD-1 were further validated by western blot analysis, biolayer interferometry, and flow cytometry. Structural analysis showed that the epitope is partially within the binding interface of PD-1 and PD-L1, and this epitope also partially overlaps with that of nivolumab and pembrolizumab. These results demonstrate that sintilimab can attenuate PD-1 activation by directly competing with the interaction between PD-1 and PD-L1 through binding with the key residues of the FG loop on PD-1. This study also demonstrates the high efficiency and accuracy of AbMap for determining the binding epitope of therapeutic antibodies.

scholarly journals A Critical Factor for Quantifying Proteins in Unmodified Gold Nanoparticles-Based Aptasensing: The Effect of pH

Chemosensors ◽  
2020 ◽  
Vol 8 (4) ◽  
pp. 98
Author(s):  
Dai Lu ◽  
Dong Zhang ◽  
Qian Zhao ◽  
Xiangyang Lu ◽  
Xingbo Shi
Keyword(s):  
Gold Nanoparticles ◽  
Limit Of Detection ◽  
Protein Detection ◽  
Critical Factor ◽  
Cost Effective ◽  
Protein Corona ◽  
Target Protein ◽  
Universal Method ◽  
Salt Addition ◽  
Positively Charged

Unmodified gold nanoparticles (AuNPs)-based aptasensing (uGA) assay has been widely implemented in the determination of many different targets, but there are few reports on protein detection using uGA. Here, we designed a uGA assay for protein detection including the elimination of interfering proteins. Positively charged protein can be absorbed directly on the surface of AuNPs to form “protein corona”, which results in the aggregation of AuNPs even without salt addition, thereby preventing target protein detection. To overcome this problem, we systematically investigated the effect of modifying the pH of the solution during the uGA assay. A probe solution with a pH slightly higher than the isoelectric points (pI) of the target protein was optimal for protein detection in the uGA assay, allowing the aptamer to selectively detect the target protein. Three proteins (beta-lactoglobulin, lactoferrin, and lysozyme) with different pI were chosen as model proteins to validate our method. Positively charged interfering proteins (with pIs higher than the optimal pH) were removed by centrifugation of protein corona/AuNPs aggregates before the implementation of actual sample detection. Most importantly, the limit of detection (LOD) for all three model proteins was comparable to that of other methods, indicating the significance of modulating the pH. Moreover, choosing a suitable pH for a particular target protein was validated as a universal method, which is significant for developing a novel, simple, cost-effective uGA assay for protein detection.

scholarly journals Colorimetric detection of protein via the terminal protection of small-molecule-linked DNA and unmodified gold nanoparticles

RSC Advances ◽  
2018 ◽  
Vol 8 (68) ◽  
pp. 38758-38764
Author(s):  
Jianwei Zhao ◽  
Cuiping Li ◽  
Guimin Ma ◽  
Wenhui Hao ◽  
Hongxia Jia
Keyword(s):  
Gold Nanoparticles ◽  
Small Molecule ◽  
Colorimetric Detection ◽  
Protein Detection ◽  
Target Protein

A novel colorimetric strategy for protein detection was developed based on unmodified gold nanoparticles (AuNPs) and terminal protection from a target protein.

Target Protein Identification on Photocatalyst‐Functionalized Magnetic Affinity Beads

2020 ◽  
Vol 101 (1) ◽  
Author(s):  
Michihiko Tsushima ◽  
Shinichi Sato ◽  
Keita Nakane ◽  
Hiroyuki Nakamura
Keyword(s):  
Protein Identification ◽  
Target Protein ◽  
Affinity Beads

Clickable gold-nanoparticles as generic probe precursors for facile photoaffinity labeling application

2021 ◽  
Author(s):  
Kanna Mori ◽  
Kaori Sakurai
Keyword(s):  
Gold Nanoparticles ◽  
Small Molecules ◽  
Photoaffinity Labeling ◽  
Protein Analysis ◽  
Target Protein ◽  
Photoaffinity Probes ◽  
One Step

Clickable photoreactive gold nanoparticles have been developed to facilitate one-step preparation of photoaffinity probes for bioactive small molecules and their application to target protein analysis.

From noncovalent to covalent bonds: a paradigm shift in target protein identification

2013 ◽  
Vol 9 (4) ◽  
pp. 544 ◽  
Author(s):  
Jongmin Park ◽  
Minseob Koh ◽  
Seung Bum Park
Keyword(s):  
Paradigm Shift ◽  
Protein Identification ◽  
Target Protein ◽  
Covalent Bonds

IgE-binding epitope mapping of tropomyosin allergen (Exo m 1) from Exopalaemon modestus, the freshwater Siberian prawn

2020 ◽  
Vol 309 ◽  
pp. 125603 ◽  
Author(s):  
Ziye Zhang ◽  
Xiu-Min Li ◽  
Hang Xiao ◽  
Anna Nowak-Wegrzyn ◽  
Peng Zhou
Keyword(s):  
Epitope Mapping ◽  
Ige Binding ◽  
Binding Epitope ◽  
Exopalaemon Modestus
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