Characterization of polystyrene-binding peptides (PS-tags) for site-specific immobilization of proteins

Kumada, Yoichi; Kuroki, Daisuke; Yasui, Hidefumi; Ohse, Takuhito; Kishimoto, Michimasa

doi:10.1016/j.jbiosc.2009.11.005

Cited by 38 publications

(30 citation statements)

References 19 publications

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“…The proposed mixed -sheet and disordered secondary structure character of the PS-tag (Figure 4) is advantageous for high affinity interaction with the plate surface [ K D =169 nM; (27)] due to its likely conformational flexibility and planar presentation at the plate surface. If the overall structure was more helical, fewer residues would interact with the plate surface, likely decreasing peptide affinity at the plate surface.…”

Section: Resultsmentioning

confidence: 99%

“…An N-terminal (Lys) 7 extension used for peptide immunoassays (41), which would adopt a disordered structure in solution (42), likely possesses the optimal peptide fusion characteristics (hydrophilic, positive charge, and disordered structure) for a high affinity, minimally structure-perturbing peptide extension sequence to improve immunoassays using peptides as capture agents in direct ELISAs. The PS-tag was shown to have a favorable planar representation at the plate surface (-sheet) and disordered structure, was biopanned specifically against a polystyrene surface, and has been show to be perform in ELISAs for a number of proteins (26,27,29,30), whereas the optimal length of the poly(Lys) N-terminal sequence was shown to be dependent on the length of the original peptide sequence (41). The PS-tag was further optimized to reveal that a mix of both basic residues (Arg) and hydrophobic (Ala and Ile) provided the highest surface affinity and surface coverage (26,27).…”

Section: Resultsmentioning

confidence: 99%

“…The PS-tag was shown to have a favorable planar representation at the plate surface (-sheet) and disordered structure, was biopanned specifically against a polystyrene surface, and has been show to be perform in ELISAs for a number of proteins (26,27,29,30), whereas the optimal length of the poly(Lys) N-terminal sequence was shown to be dependent on the length of the original peptide sequence (41). The PS-tag was further optimized to reveal that a mix of both basic residues (Arg) and hydrophobic (Ala and Ile) provided the highest surface affinity and surface coverage (26,27). …”

Section: Resultsmentioning

confidence: 99%

“…Recently, Kumada, et al, identified a 12-amino acid polystyrene binding sequence (PS-tag) using the Flitrx random peptide display library that could be used for improved hydrophilic polystyrene plate binding of fusion proteins (26). This PS-tag (PS191; RAFIASRRIRRP) and PS196 (RIIIRRIR), derived from the original PS191 sequence, are able to maintain polystyrene binding capacity in the presence of detergents, such as Tween-20, and BSA used during adsorption steps (27). Both the PS191 and PS196 have high affinity for polystyrene, 169 and 86 nM respectively, and show similar maximum adsorption, 81 and 86 mg/m 2 respectively, with the PS191 sequence being more hydrophilic overall, -0.792, compared with the PS196, -0.500 (28).…”

mentioning

confidence: 99%

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Increased Affinity and Solubility of Peptides Used for Direct Peptide ELISA on Polystyrene Surfaces Through Fusion with a Polystyrene-Binding Peptide Tag

Kogot

Sarkes²,

Val-Addo³

et al. 2012

BioTechniques

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Peptide reagents can serve as alternatives or replacements to antibodies in sensing or diagnostic applications. The passive adsorption of peptides onto polystyrene surfaces can limit the target binding capability, especially for short, positively charged, or hydrophobic sequences. In this report, we show that fusing a peptide with a previously characterized 12-amino acid polystyrene binding sequence (PS-tag) improves overall peptide solubility and enzyme-linked immunosorbent assay (ELISA) results using the peptide as a capture agent. Specific improvements for protective antigen (PA; Bacillus anthracis) protein binding peptides selected from bacterial surface display were compared with native or biotinylated peptides. The PS-tag was added to either peptide terminus, using a (Gly)(4) spacer, and comparable binding affinities were obtained. Fusion with the PS-tag did not have any negative impact on peptide secondary structure as measured by circular dichroism. The addition of the PS-tag provides a convenient method to utilize peptide reagents from peptide display libraries as capture agents in an ELISA format without the need for a biotin tag or concerns about passive adsorption of critical residues for target capture.

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

mentioning

confidence: 99%

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Increased Affinity and Solubility of Peptides Used for Direct Peptide ELISA on Polystyrene Surfaces Through Fusion with a Polystyrene-Binding Peptide Tag

Kogot

Sarkes²,

Val-Addo³

et al. 2012

BioTechniques

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“…Difficulties then include depency on such MS instrument that is not available in most general hospital. The next step should be the development of a peptide-ELISA protocol to detect the DAK fragment LLSKLSVLLLEKMG in the serum [29,30]. …”

Section: Discussionmentioning

confidence: 99%

Serum dihydroxyacetone kinase peptide m/z 520.3 as predictor of disease severity in patients with compensated chronic hepatitis B

Jia

et al. 2013

J Transl Med

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Background & aimDue to known limitations of liver biopsy, reliable non-invasive serum biomarkers for chronic liver diseases are needed. We performed serum peptidomics for such investigation in compensated chronic hepatitis B (CHB) patients.MethodsLiquid chromatography combined with tandem mass spectrometry (LC-MS/MS) was used to identify differentially expressed peptides in sera from 40 CHB patients (20 with S0G0-S1G1 and 20 with S3G3-S4G4). Ion pair quantification from differentially expressed peptides in a validation set of sera from 86 CHB patients was done with multiple reaction monitoring (MRM).Results21 differentially represented peptide peaks were found through LC-MS/MS. Ion pairs generated from eleven of these peptides (m/z < 800) were quantified by MRM. Summed peak area ratios of 6 ion pairs from peptide m/z 520.3 (176.1, 353.7, 459.8, 503.3, 351.3, 593.1), which was identified as dihydroxyacetone kinase (DAK) fragment, decreased from mild to advanced stages of fibrosis or inflammation. Area Under Receiver Operating Characteristic Curves (AUROCs) of five ion models discriminating fibrosis degrees were 0.871 ~ 0.915 (S2-4 versus S0-1) and 0.804 ~ 0.924 (S3-4 versus S0-2). AUROCs discriminating inflammation grades were 0.840 ~ 0.902 (G2-4 versus G0-1) and 0.787 ~ 0.888 (G3-4 versus G0-2). The diagnostic power of these models provides improved sensitivity and specificity for predicting disease progression as compared to aspartate aminotransferase to platelet ratio index (APRI), FIB-4, Forn’s index and serum DAK protein.ConclusionsThe peptide fragment (m/z 520.3) of DAK is a promising biomarker to guide timing of antiviral treatment and to avoid liver biopsy in compensated CHB patients.

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Directed evolution of polypropylene and polystyrene binding peptides

Rübsam

Weber

Jakob

et al. 2017

Biotech & Bioengineering

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Surface functionalization of biological inert polymers (e.g., polypropylene PP; polystyrene PS) with material binding peptides facilitates an efficient immobilization of enzymes, bioactive peptides or antigens at ambient temperature in water. The developed robust directed evolution protocol enables to tailor polymer binding anchor peptides (PBPs) for efficient binding under application conditions. Key for a successful directed evolution campaign was to develop an epPCR protocol with a very high mutation frequency (60 mutations/kb) to ensure sufficient diversity in PBPs (47 aas LCI: “liquid chromatography peak I”; 44 aas TA2: “Tachystatin A2”). LCI and TA2 were genetically fused to the reporter egfp to quantify peptide binding on PP and PS by fluorescence analysis. The Peptide‐Polymer evolution protocol (PePevo protocol) was validated in two directed evolution campaigns for two PBPs and polymers (LCI: PP; TA2: PS). Surfactants were used as selection pressure for improved PBP binders (non‐ionic surfactant Triton X‐100; 1 mM for LCI‐PP // anionic surfactant LAS; 0.5 mM for TA2‐PS). PePevo yielded an up to three fold improved PP‐binder (LCI‐M1‐PP: I24T, Y29H, E42 K and LCI‐M2‐PP: D31V, E42G) and an up to six fold stronger PS‐binder (TA2‐M1‐PS: R3S, L6P, V12 K, S15P, C29R, R30L, F33S, Y44H and TA2‐M2‐PS: F9C, C24S, G26D, S31G, C41S, Y44Q).

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Characterization of polystyrene-binding peptides (PS-tags) for site-specific immobilization of proteins

Cited by 38 publications

References 19 publications

Increased Affinity and Solubility of Peptides Used for Direct Peptide ELISA on Polystyrene Surfaces Through Fusion with a Polystyrene-Binding Peptide Tag

Increased Affinity and Solubility of Peptides Used for Direct Peptide ELISA on Polystyrene Surfaces Through Fusion with a Polystyrene-Binding Peptide Tag

Serum dihydroxyacetone kinase peptide m/z 520.3 as predictor of disease severity in patients with compensated chronic hepatitis B

Directed evolution of polypropylene and polystyrene binding peptides

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