Cellular analyses for label-free and rapid HER2-positive cancer diagnosis based on SPRi-modified with nanobody

Chen, Ribao; Li, Qin; Zeng, Youjun; Sang, Wei; Tang, Shuo; Jiao, Yue; Zhou, Jie; Xu, Zhourui; Shao, Yonghong; Ying, Ming; Xu, Gaixia

doi:10.1016/j.snb.2023.134490

Cited by 4 publications

(1 citation statement)

References 31 publications

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“…They can also be efficiently expressed in prokaryotic systems, facilitating cost-effective mass production. These inherent advantages render them particularly favorable for research and application in various fields such as analytical chemistry, disease diagnosis, and therapy . In recent years, the increasing number of researchers have harnessed the potential of Nbs in developing highly sensitive immunoassays targeting various entities including food allergens, mycotoxins, pesticides, and foodborne pathogens .…”

Section: Introductionmentioning

confidence: 99%

Enhancing Nanobody Immunoassays through Ferritin Fusion: Construction of a Salmonella-Specific Fenobody for Improved Avidity and Sensitivity

Liao,

Wang,

Guo

et al. 2024

J. Agric. Food Chem.

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Nanobodies (Nbs) serve as powerful tools in immunoassays. However, their small size and monovalent properties pose challenges for practical application. Multimerization emerges as a significant strategy to address these limitations, enhancing the utilization of nanobodies in immunoassays. Herein, we report the construction of a Salmonella-specific fenobody (Fb) through the fusion of a nanobody to ferritin, resulting in a self-assembled 24-valent nanocage-like structure. The fenobody exhibits a 35-fold increase in avidity compared to the conventional nanobody while retaining good thermostability and specificity. Leveraging this advancement, three ELISA modes were designed using Fb as the capture antibody, along with unmodified Nb422 (FbNb-ELISA), biotinylated Nb422 (FbBio-ELISA), and phage-displayed Nb422 (FbP-ELISA) as the detection antibody, respectively. Notably, the FbNb-ELISA demonstrates a detection limit (LOD) of 3.56 × 10 4 CFU/mL, which is 16-fold lower than that of FbBio-ELISA and similar to FbP-ELISA. Moreover, a fenobody and nanobody sandwich chemiluminescent enzyme immunoassay (FbNb-CLISA) was developed by replacing the TMB chromogenic substrate with luminal, resulting in a 12-fold reduction in the LOD. Overall, the ferritin-displayed technology represents a promising methodology for enhancing the detection performance of nanobody-based sandwich ELISAs, thereby expanding the applicability of Nbs in food detection and other fields requiring multivalent modification.

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