Recombinant GST-I-Aβ28-induced efficient serum antibody against Aβ42

Huang, Xuemei; Wang, Jiapeng; Cui, Lili; Zou, Xiaohuan; Zhang, Yingjiu

doi:10.1016/j.jneumeth.2009.10.026

Cited by 14 publications

(8 citation statements)

References 24 publications

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“…Aβ42 was purchased from Sigma-Aldrich (Sigma-Aldrich, USA),from which the various Aβ42 aggregates were prepared and confirmed by electron microscopy as described previously [14,15].…”

Section: Preparation Of Aβ42 Aggregatesmentioning

confidence: 99%

“…Then the cytotoxicity-neutralizing or cytotoxicityinhibiting activities of the selected scFv were assessed as described previously with little modification [14,15]. Briefly, the four Aβ42 species (10 μM final concentration) and the selected scFv (10 μM final concentration) were added simultaneously or sequentially into the above wells.…”

Section: Estimation Of the Protective Effects Of Scfv As Against Aβ42mentioning

confidence: 99%

“…Many researchers have recently found that different fragments of Aβ42 peptide displayed different cytotoxic activities [12,13] and that different antibodies obtained by immunization with different fragments of Aβ42 peptide exhibited different reactivities and specificities to Aβ42 aggregates [14,15]. Moreover, researchers have also shown that Aβ42 oligomers as well as protofibrils in general have strong neurotoxicity and are the initial and pivotal toxins responsible for AD occurrence and development [16,17].…”

Section: Introductionmentioning

confidence: 99%

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Functional Characteristics and Molecular Mechanism of a New scFv Antibody Against Aβ42 Oligomers and Immature Protofibrils

et al. 2014

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Amyloid β peptide (Aβ42) is a major determinant of Alzheimer's disease (AD). In this study, we studied a novel single-chain variable fragment (scFv), AS, generated from an antibody library of AD patients, which recognized and bound specifically to medium-size amyloid β peptide (Aβ42) oligomers and immature protofibrils (25-55 kDa) and, more importantly, reduced their level by blocking their formation or inducing their disassembly. Consequently, scFv AS ameliorated or prevented their cytotoxicity and protected SH-SY5Y cells and primary cultured neurons in vitro from their damage in a concentration-dependent manner. Comparison of its cytotoxicity-inhibiting and cytotoxicity-neutralizing activities indicated that scFv AS displayed its protective effect on target cells mainly due to its cytotoxicity-inhibitory activity though it could also neutralize the cytotoxicity. We also found that scFv AS could efficiently cross the in vitro BBB model with a delivery efficiency of over 70% after a 60-min post-administration. The scFv AS was a monovalent antibody with an affinity constant (KD) of 5.5 × 10(-6) M and a binding threshold of 6.25 × 10(-4) μM for Aβ42 oligomers. The molecular docking simulations of Aβ42 to scFv AS revealed that scFv AS tends to approached Aβ42 oligomers and immature protofibrils mainly by their hydrophobic interaction and then drew Aβ42 molecule into the gap between VL and VH domains of scFv AS by hydrophilic interaction between scFv AS and the N-terminal region (residues 1-15) of Aβ42 and the hydrophobic interactions between scFv AS and the middle region (residues 20-33) of Aβ42. The combination of scFv AS with Aβ42 was realized likely through an induced-fit process.

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“…Aβ42 was purchased from Sigma-Aldrich (Sigma-Aldrich, USA),from which the various Aβ42 aggregates were prepared and confirmed by electron microscopy as described previously [14,15].…”

Section: Preparation Of Aβ42 Aggregatesmentioning

confidence: 99%

Section: Estimation Of the Protective Effects Of Scfv As Against Aβ42mentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Functional Characteristics and Molecular Mechanism of a New scFv Antibody Against Aβ42 Oligomers and Immature Protofibrils

et al. 2014

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“…The structure-toxicity relationship of Aβ42 aggregates was further revealed by serum antibodies induced by Aβ1–9, Aβ1–28, and Aβ42 in our previous studies ( Figure 3 ) [ 33 , 34 ]. It has been reported that the serum antibodies induced by (Aβ9)16 (16 tandem repeats of Aβ1–9) display a high immunoreactivity to Aβ42M and Aβ42O ( p < 0.01, Aβ42M/Aβ42O and Aβ42P/Aβ42F) but a low immunoreactivity to Aβ42P ( p < 0.05 compared with pre-serum) and no immunoreactivity to Aβ42 mature fibrils ( p > 0.05 compared with pre-serum) ( Figure 3 A).…”

Section: A Specific Integrated Conformation Underlies the Neurotoxici...mentioning

confidence: 73%

Conformational Essentials Responsible for Neurotoxicity of Aβ42 Aggregates Revealed by Antibodies against Oligomeric Aβ42

Song

Zhang²,

Zhang³

2022

Molecules

Self Cite

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Soluble aggregation of amyloid β-peptide 1-42 (Aβ42) and deposition of Aβ42 aggregates are the initial pathological hallmarks of Alzheimer’s disease (AD). The bipolar nature of Aβ42 molecule results in its ability to assemble into distinct oligomers and higher aggregates, which may drive some of the phenotypic heterogeneity observed in AD. Agents targeting Aβ42 or its aggregates, such as anti-Aβ42 antibodies, can inhibit the aggregation of Aβ42 and toxicity of Aβ42 aggregates to neural cells to a certain extent. However, the epitope specificity of an antibody affects its binding affinity for different Aβ42 species. Different antibodies target different sites on Aβ42 and thus elicit different neuroprotective or cytoprotective effects. In the present review, we summarize significant information reflected by anti-Aβ42 antibodies in different immunotherapies and propose an overview of the structure (conformation)−toxicity relationship of Aβ42 aggregates. This review aimed to provide a reference for the directional design of antibodies against the most pathogenic conformation of Aβ42 aggregates.

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“…Because protein fragments are more amenable to engineering and production, the hunt for the smallest antibody fragment capable of binding to antigens continues and progressions such as 55-kDa Fab and 28-kDa scFv have been made. However, problems remain in the expression and aggregation of these antibodies [8,9]. A distinct type of antibody, which contains only the heavy chain, was discovered in the serum of alpaca, camel and cartilage fish in 1993 [10].…”

Section: Introductionmentioning

confidence: 99%

Oriented Immobilization and Quantitative Analysis Simultaneously Realized in Sandwich Immunoassay via His-Tagged Nanobody

Cao

Huang

et al. 2019

Molecules

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Despite the advantages of the nanobody, the unique structure limits its use in sandwich immunoassay. In this study, a facile protocol of sandwich immunoassay using the nanobody was established. In brief, β amyloid and SH2, an anti-β amyloid nanobody, were used as capture antibody and antigen, respectively. The SH2 fused with His-tag was first purified and absorbed on Co2+-NTA functional matrix and then immobilized through H2O2 oxidation of Co2+ to Co3+ under the optimized conditions. Then, 150 mM imidazole and 20 mM EDTA were introduced to remove the unbound SH2. The immobilized SH2 showed highly-sensitive detection of β amyloid. It is interesting that the quantification of the sandwich immunoassay was carried out by determining the His-tag of the detection nanobody, without interference from the His-tag of the capture nanobody. The immobilized SH2 detached exhibited outstanding stability during 30 days of storage. Taken together, His6-tag facilitated both the oriented immobilization of capture antibody and quantitative assay of detection antibody in sandwich immunoassay. We propose a facile and efficient sandwich immunoassay method that opens new avenue to the study of His-tagged protein interactions.

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Recombinant GST-I-Aβ28-induced efficient serum antibody against Aβ42

Cited by 14 publications

References 24 publications

Functional Characteristics and Molecular Mechanism of a New scFv Antibody Against Aβ42 Oligomers and Immature Protofibrils

Functional Characteristics and Molecular Mechanism of a New scFv Antibody Against Aβ42 Oligomers and Immature Protofibrils

Conformational Essentials Responsible for Neurotoxicity of Aβ42 Aggregates Revealed by Antibodies against Oligomeric Aβ42

Oriented Immobilization and Quantitative Analysis Simultaneously Realized in Sandwich Immunoassay via His-Tagged Nanobody

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