Chemical modification of M13 bacteriophage as nanozyme container for dramatically enhanced sensitivity of colorimetric immunosensor

Fang, Hao; Zhan, Shengnan; Feng, Lei; Chen, Xuelan; Guo, Qian; Guo, Yuqian; He, Qinghua; Xiong, Yonghua

doi:10.1016/j.snb.2021.130368

Cited by 28 publications

(14 citation statements)

References 32 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…In our previous work, an M13 OTA bacteriophage was panned from a commercial phage display seven-peptide library, where the OTA mimotope peptide sequence of “GMSWMMA” was fused on the p3 proteins to endow the phage selectivity toward anti-OTA mAb [ 29 ]. The titer of the amplified M13 OTA was tested with a plate count method [ 28 ]. As shown in Figure S1 , 10 μL of 10 8 -fold diluted phage solution developed 340 plaques of infected E. coli cells on the plates, indicating that the stock solution had a titer of 5.6 × 10 11 pfu/mL.…”

Section: Resultsmentioning

confidence: 99%

“…Through a gene modification-fusion expression process, mimotopes can be integrated on the N-terminals of p3 proteins; this process endows an M13 phage with mimicking ability for target antigen [ 22 , 23 , 24 ]. In addition, the detection sensitivity can be amplified by extensively functionalizing the abundant copies of major p8 proteins as the high-capacity containers of signal transducers or regulators (e.g., fluorescent dyes and enzymes) [ 25 , 26 , 27 , 28 ]. For example, Fang et al [ 28 ] adopted a sulfhydryl-modified M13 phage as high-density container of Au@Ag nanozymes to enhance the sensitivity of the colorimetric biosensor.…”

Section: Introductionmentioning

confidence: 99%

“…In addition, the detection sensitivity can be amplified by extensively functionalizing the abundant copies of major p8 proteins as the high-capacity containers of signal transducers or regulators (e.g., fluorescent dyes and enzymes) [ 25 , 26 , 27 , 28 ]. For example, Fang et al [ 28 ] adopted a sulfhydryl-modified M13 phage as high-density container of Au@Ag nanozymes to enhance the sensitivity of the colorimetric biosensor.…”

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Dramatically Enhancing the Sensitivity of Immunoassay for Ochratoxin A Detection by Cascade-Amplifying Enzyme Loading

Song

Feng

Leng

et al. 2021

Toxins

Self Cite

View full text Add to dashboard Cite

Enzyme-linked immunosorbent assay (ELISA) is widely used in the routine screening of mycotoxin contamination in various agricultural and food products. Herein, a cascade-amplifying system was introduced to dramatically promote the sensitivity of an immunoassay for ochratoxin A (OTA) detection. Specifically, a biotinylated M13 bacteriophage was introduced as a biofunctional competing antigen, in which a seven-peptide OTA mimotope fused on the p3 protein of M13 was used to specifically recognize an anti-OTA monoclonal antibody, and the biotin molecules modified on capsid p8 proteins were used in loading numerous streptavidin-labeled polymeric horseradish peroxidases (HRPs). Owing to the abundance of biotinylated p8 proteins in M13 and the high molar ratio between HRP and streptavidin in streptavidin-polyHRP, the loading amount of HRP enzymes on the M13 bacteriophage were greatly boosted. Hence, the proposed method exhibited high sensitivity, with a limit of detection of 2.0 pg/mL for OTA detection, which was 250-fold lower than that of conventional ELISA. In addition, the proposed method showed a slight cross-reaction of 2.3% to OTB, a negligible cross-reaction for other common mycotoxins, and an acceptable accuracy for OTA quantitative detection in real corn samples. The practicability of the method was further confirmed with a traditional HRP-based ELISA method. In conclusion, the biotinylated bacteriophage and polyHRP structure showed potential as a cascade-amplifying enzyme loading system for ultra-trace OTA detemination, and its application can be extended to the detection of other analytes by altering specific mimic peptide sequences.

show abstract

Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Dramatically Enhancing the Sensitivity of Immunoassay for Ochratoxin A Detection by Cascade-Amplifying Enzyme Loading

Song

Feng

Leng

et al. 2021

Toxins

Self Cite

View full text Add to dashboard Cite

show abstract

“…The concentration of the amplified M13 OTA phage was determined via the plate count method [30]. Briefly, a swatch of ER2738 cells grown on an LB/Tet plate (LB plate containing 10 µg/mL of tetracycline) was inoculated into 5-10 mL of LB medium and incubated under shaking for 6-10 h to mid-log phase (OD600 = 0.5).…”

Section: Propagation Of M13 Ota Bacteriophagementioning

confidence: 99%

“…For instance, Lee et al [29] adopted the M13 phage as a biological template to amplify the surface-enhanced Raman scattering signal for protein sensing. Fang et al [30] used a sulfydryl-modified M13 phage as an Au-Ag nanozyme container to improve the sensitivity of a colorimetric biosensor.…”

Section: Introductionmentioning

confidence: 99%

Eco-Friendly Fluorescent ELISA Based on Bifunctional Phage for Ultrasensitive Detection of Ochratoxin A in Corn

Tong

Fang

Xiong

et al. 2021

Foods

Self Cite

View full text Add to dashboard Cite

Conventional enzyme-linked immunosorbent assay (ELISA) is commonly used for Ochratoxin A (OTA) screening, but it is limited by low sensitivity and harmful competing antigens of enzyme-OTA conjugates. Herein, a bifunctional M13 bacteriophage with OTA mimotopes fused on the p3 protein and biotin modified on major p8 proteins was introduced as an eco-friendly competing antigen and enzyme container for enhanced sensitivity. Mercaptopropionic acid-modified quantum dots (MPA-QDs), which are extremely sensitive to hydrogen peroxide, were chosen as fluorescent signal transducers that could manifest glucose oxidase-induced fluorescence quenching in the presence of glucose. On these bases, a highly sensitive and eco-friendly fluorescent immunoassay for OTA sensing was developed. Under optimized conditions, the proposed method demonstrates a good linear detection of OTA from 4.8 to 625 pg/mL and a limit of detection (LOD) of 5.39 pg/mL. The LOD is approximately 26-fold lower than that of a conventional horse radish peroxidase (HRP) based ELISA and six-fold lower than that of a GOx-OTA conjugate-based fluorescent ELISA. The proposed method also shows great specificity and accepted accuracy for analyzing OTA in real corn samples. The detection results are highly consistent with those obtained using the ultra-performance liquid chromatography-fluorescence detection method, indicating the high reliability of the proposed method for OTA detection. In conclusion, the proposed method is an excellent OTA screening platform over a conventional ELISA and can be easily extended for sensing other analytes by altering specific mimic peptide sequences in phages.

show abstract

Exploring Nanozymes for Organic Substrates: Building Nano‐organelles

Liu,

Gao,

Qin

et al. 2024

Angewandte Chemie

View full text Add to dashboard Cite

Since the discovery of the first peroxidase nanozyme (Fe3O4), numerous nanomaterials have been reported to exhibit intrinsic enzyme‐like activity toward inorganic oxygen species, such as H2O2, oxygen, and O2•‐. However, the exploration of nanozymes targeting organic compounds holds transformative potential in the realm of industrial synthesis. This review provides a comprehensive overview of the diverse types of nanozymes that catalyze reactions involving organic substrates and discusses their catalytic mechanisms, structure‐activity relationships, and methodological paradigms for discovering new nanozymes. Additionally, we propose a forward‐looking perspective on designing nanozyme formulations to mimic subcellular organelles, such as chloroplasts, termed “nano‐organelles”. Finally, we analyze the challenges encountered in nanozyme synthesis, characterization, nano‐organelle construction and applications while suggesting directions to overcome these obstacles and enhance nanozyme research in the future. Through this review, our goal is to inspire further research efforts and catalyze advancements in the field of nanozymes, fostering new insights and opportunities in chemical synthesis.

show abstract

Chemical modification of M13 bacteriophage as nanozyme container for dramatically enhanced sensitivity of colorimetric immunosensor

Cited by 28 publications

References 32 publications

Dramatically Enhancing the Sensitivity of Immunoassay for Ochratoxin A Detection by Cascade-Amplifying Enzyme Loading

Dramatically Enhancing the Sensitivity of Immunoassay for Ochratoxin A Detection by Cascade-Amplifying Enzyme Loading

Eco-Friendly Fluorescent ELISA Based on Bifunctional Phage for Ultrasensitive Detection of Ochratoxin A in Corn

Exploring Nanozymes for Organic Substrates: Building Nano‐organelles

Contact Info

Product

Resources

About