A rapid, easy, and sensitive method for detecting His-tag-containing chitinase based on ssDNA aptamers and gold nanoparticles

Cao, Zhuoning; Wang, Sai; Liu, Zhen; Xue, Changhu; Mao, Xiangzhao

doi:10.1016/j.foodchem.2020.127230

Cited by 8 publications

(1 citation statement)

References 22 publications

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“…Also, by combining DNA aptamer and gold nanoparticle technologies, quick and qualitative determination of His-tagged protein was reported. However, their working range is narrow [ 6 , 7 ]. This is not conducive to knowing when to harvest the target recombinant products in large-scale production, which may result in a decreased yield due to premature recovery or protein degradation.…”

Section: Introductionmentioning

confidence: 99%

Construction of Fluorescent Immunosensor Quenchbody to Detect His-Tagged Recombinant Proteins Produced in Bioprocess

Ning

Yasuda

Kitaguchi

et al. 2021

Sensors

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With the widespread application of recombinant DNA technology, many useful substances are produced by bioprocesses. For the monitoring of the recombinant protein production process, most of the existing technologies are those for the culture environment (pH, O2, etc.). However, the production status of the target protein can only be known after the subsequent separation and purification process. To speed up the monitoring of the production process and screening of the higher-yield target protein variants, here we developed an antibody-based His-tag sensor Quenchbody (Q-body), which can quickly detect the C-terminally His-tagged recombinant protein produced in the culture medium. Compared with single-chain Fv-based Q-body having one dye, the Fab-based Q-body having two dyes showed a higher response. In addition, not only was fluorescence response improved but also detection sensitivity by the mutations of tyrosine to tryptophan in the heavy chain CDR region. Moreover, the effect of the mutations on antigen-binding was successfully validated by molecular docking simulation by CDOCKER. Finally, the constructed Q-body was successfully applied to monitor the amount of anti-SARS CoV-2 nanobody secreted into the Brevibacillus culture media.

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Section: Introductionmentioning

confidence: 99%