Identification and characterization of peptide fragments for the direct and site-specific immobilization of functional proteins onto the surface of silicon nitride

Kumada, Yoichi; Ootsuka, Takeru; Asada, Masashi; Yoshizuka, Saori; Chiyama, Masateru; Sakane, Masayasu; Fida, Hasan M.; Sawada, Kazuaki; Okumura, Kōichi; Kishimoto, Michimasa

doi:10.1016/j.jbiotec.2014.04.013

Cited by 10 publications

(4 citation statements)

References 24 publications

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“…As an alternative method that is free of the above mentioned shortcomings, we and another group used cellular protein libraries to select material‐binding proteins. In this method, a target material was first mixed with a protein mixture of bacterial cell lysate.…”

Section: Material‐binding Peptidesmentioning

confidence: 99%

“…In another study, binding peptides were directly selected from a peptide library prepared by trypsin digestion of the binding protein . Kumada et al successfully isolated peptides binding to the surface of hydrophilic polystyrene , polycarbonate , poly(methylmethacrylate) , and silicon nitride (SiN) from the amino acid sequences of E. coli proteins.…”

Section: Material‐binding Peptidesmentioning

confidence: 99%

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Rapid on‐site detection of airborne asbestos fibers and potentially hazardous nanomaterials using fluorescence microscopy‐based biosensing

Kuroda

Alexandrov

Nishimura

et al. 2016

Biotechnology Journal

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A large number of peptides with binding affinity to various inorganic materials have been identified and used as linkers, catalysts, and building blocks in nanotechnology and nanobiotechnology. However, there have been few applications of material-binding peptides in the fluorescence microscopy-based biosensing (FM method) of environmental pollutants. A notable exception is the application of the FM method for the detection of asbestos, a dangerous industrial toxin that is still widely used in many developing countries. This review details the selection and isolation of asbestos-binding proteins and peptides with sufficient specificity to distinguish asbestos from a large variety of safer fibrous materials used as asbestos substitutes. High sensitivity to nanoscale asbestos fibers (30-35 nm in diameter) invisible under conventional phase contrast microscopy can be achieved. The FM method is the basis for developing an automated system for asbestos biosensing that can be used for on-site testing with a portable fluorescence microscope. In the future, the FM method could also become a useful tool for detecting other potentially hazardous nanomaterials in the environment.

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Section: Material‐binding Peptidesmentioning

confidence: 99%

Section: Material‐binding Peptidesmentioning

confidence: 99%

Rapid on‐site detection of airborne asbestos fibers and potentially hazardous nanomaterials using fluorescence microscopy‐based biosensing

Kuroda

Alexandrov

Nishimura

et al. 2016

Biotechnology Journal

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“…Thus, peptides binding to PS, PC, PMMA, and silicon nitride (SiN) have been successfully identified. Genetic fusion of these peptides to the C‐terminal region of a model enzyme, glutathione S‐transferase (GST), resulted in site‐directed immobilization with higher density and maintenance of high enzymatic activity on the surfaces of target materials …”

Section: Introductionmentioning

confidence: 99%

“…Genetic fusion of these peptides to the C-terminal region of a model enzyme, glutathione S-transferase (GST), resulted in site-directed immobilization with higher density and maintenance of high enzymatic activity on the surfaces of target materials. [9][10][11][12][13] Furthermore, these peptides were applicable to the immobilization of recombinant antibody fragments. For example, PS-tag-fused scFvs and Fabs with high antigen-binding activity were successfully immobilized by novel solid-phase refolding methods that denatured antibody fragments obtained from inclusion bodies were directly immobilized on the surface of a hydrophilic PS (phi-PS) plate under semidenatured conditions.…”

Section: Introductionmentioning

confidence: 99%

Efficient preparation and site‐directed immobilization of VHH antibodies by genetic fusion of poly(methylmethacrylate)‐binding peptide (PMMA‐Tag)

Kumada

Kang

Yamakawa

et al. 2015

Biotechnology Progress

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A PMMA-binding peptide (PMMA-tag) was genetically fused with the C-terminal region of an anti-human chorionic gonadotropin (hCG) single-domain antibody (VHH). It was over-expressed in an insoluble fraction of E. coli cells, and recovered in the presence of 8 M urea via one-step IMAC purification. Monomeric and denatured PMMA-tag-fused VHH (VHH-PM) was successfully prepared via the reduction and oxidation of VHH-PM at a concentration less than 1 mg/mL in the presence of 8 M of urea. Furthermore, the VHH-PM was refolded with a recovery of more than 95% by dialysis against 50 mM TAPS at pH 8.5, because the genetic fusion of PMMA-tag resulted in a decrease in the apparent isoelectric point (pI) of the fusion protein, and its solubility at weak alkaline pH was considerably increased. The antigen-binding activities of VHH-PM in the adsorptive state were 10-fold higher than that of VHH without a PMMA-tag. The density of VHH-PM on a PMMA plate was twice that of VHH, indicating that the site-directed attachment of a PMMA-tag resulted in positive effects to the adsorption amount as well as to the orientation of VHH-PM in its adsorptive state. The preparation and immobilization methods for VHH-PM against hCG developed in the present study were further applied to VHH-PMs against four different antigens, and consequently, those antigens with the concentrations lower than 1 ng/mL were detected by the sandwich ELISA. Thus, the VHH-PMs developed in the present study are useful for preparation of high-performance and economical immunosorbent for detection of biomarkers.

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Oriented immobilization of proteins on solid supports for use in biosensors and biochips: a review

2015

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Identification and characterization of peptide fragments for the direct and site-specific immobilization of functional proteins onto the surface of silicon nitride

Cited by 10 publications

References 24 publications

Rapid on‐site detection of airborne asbestos fibers and potentially hazardous nanomaterials using fluorescence microscopy‐based biosensing

Rapid on‐site detection of airborne asbestos fibers and potentially hazardous nanomaterials using fluorescence microscopy‐based biosensing

Efficient preparation and site‐directed immobilization of VHH antibodies by genetic fusion of poly(methylmethacrylate)‐binding peptide (PMMA‐Tag)

Oriented immobilization of proteins on solid supports for use in biosensors and biochips: a review

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