Covalently Immobilized Regenerable Immunoaffinity Layer with Orientation-Controlled Antibodies Based on Z-Domain Autodisplay

A regenerable immunoaffinity layer comprising covalently immobilized orientation-controlled antibodies was developed for use in a surface plasmon resonance (SPR) biosensor. For antibody orientation control, antibody-binding Z-domain-autodisplaying Escherichia coli (E. coli) cells and their outer membrane (OM) were utilized, and a disuccinimidyl crosslinker was employed for covalent antibody binding. To fabricate the regenerable immunoaffinity layer, capture antibodies were bound to autodisplayed Z-domains, and then treated with the crosslinker for chemical fixation to the Z-domains. Various crosslinkers, namely disuccinimidyl glutarate (DSG), disuccinimidyl suberate (DSS) and poly (ethylene glycol)-ylated bis (sulfosuccinimidyl)suberate (BS(PEG)5), were evaluated, and DSS at a concentration of 500 μM was confirmed to be optimal. The E. coli-cell-based regenerable HRP immunoassay was evaluated employing three sequential HRP treatment and regeneration steps. Then, the Oms of E. coli cells were isolated and layered on a microplate and regenerable OM-based HRP immunoassaying was evaluated. Five HRP immunoassays with four regeneration steps were found to be feasible. This regenerable, covalently immobilized, orientation-controlled OM-based immunoaffinity layer was applied to an SPR biosensor, which was capable of quantifying C-reactive protein (CRP). Five regeneration cycles were repeated using the demonstrated immunoaffinity layer with a signal difference of <10%.

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Effect of Poly(ethylene glycol), Tetramethylammonium Hydroxide, and Other Surfactants on Enhancing Performance in a Latex Particle Immunoassay of C-Reactive Protein

Analytical Chemistry ◽

10.1021/ac001530g ◽

2001 ◽

Vol 73 (14) ◽

pp. 3426-3431 ◽

Cited By ~ 13

Author(s):

Peter Holownia ◽

Soledad Perez-Amodio ◽

Christopher P. Price

Keyword(s):

Ethylene Glycol ◽

Latex Particle ◽

Tetramethylammonium Hydroxide ◽

C Reactive Protein ◽

Poly Ethylene Glycol ◽

Reactive Protein ◽

Poly Ethylene

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Thiazolidinedione (Pioglitazone) Blocks P. gingivalis- and F. nucleatum, but not E. coli, Lipopolysaccharide (LPS)-induced Interleukin-6 (IL-6) Production in Adipocytes

Journal of Dental Research ◽

10.1177/154405910508400306 ◽

2005 ◽

Vol 84 (3) ◽

pp. 240-244 ◽

Cited By ~ 9

Author(s):

M. Yamaguchi ◽

F. Nishimura ◽

H. Naruishi ◽

Y. Soga ◽

S. Kokeguchi ◽

...

Keyword(s):

Interleukin 6 ◽

Elevated Level ◽

Periodontal Pathogens ◽

C Reactive Protein ◽

E Coli ◽

Reactive Protein ◽

Periodontal Bacteria ◽

Obese Subjects ◽

Culture Supernatants ◽

Inflammatory Effect

An elevated level of C-reactive protein (CRP) predicts the future development of coronary heart disease. Periodontitis appears to up-regulate CRP. CRP is produced by hepatocytes in response to interleukin-6 (IL-6). A major source of IL-6 in obese subjects is adipocytes. We hypothesized that lipopolysaccharide (LPS) from periodontal pathogens stimulated adipocytes to produce IL-6, and that the production was suppressed by the drugs targeted against insulin resistance, thiazolidinedione (pioglitazone), since this agent potentially showed an anti-inflammatory effect. Mouse 3T3-L1 adipocytes were stimulated with E. coli, P. gingivalis, and F. nucleatum LPS. The IL-6 concentration in culture supernatants was measured. All LPS stimulated adipocytes to produce IL-6. Although pioglitazone changed adipocyte appearance from large to small, and completely suppressed P. gingivalis and F. nucleatum LPS-induced IL-6 production, E. coli LPS-induced IL-6 production was not efficiently blocked. Thus, pioglitazone completely blocked periodontal-bacteria-derived LPS-induced IL-6 production in adipocytes, a major inducer of CRP.

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A Poly(ethylene glycol)-Supported Quaternary Ammonium Salt for Highly Efficient and Environmentally Friendly Chemical Fixation of CO2 with Epoxides under Supercritical Conditions.

ChemInform ◽

10.1002/chin.200623113 ◽

2006 ◽

Vol 37 (23) ◽

Author(s):

Ya Du ◽

Jin-Quan Wang ◽

Jian-Yu Chen ◽

Fei Cai ◽

Jie-Sheng Tian ◽

...

Keyword(s):

Ethylene Glycol ◽

Ammonium Salt ◽

Quaternary Ammonium ◽

Quaternary Ammonium Salt ◽

Environmentally Friendly ◽

Chemical Fixation ◽

Poly Ethylene Glycol ◽

Supercritical Conditions ◽

Highly Efficient ◽

Poly Ethylene

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Localization and Therapy of Urinary Tract Infections of Childhood

PEDIATRICS ◽

10.1542/peds.63.3.467 ◽

1979 ◽

Vol 63 (3) ◽

pp. 467-474 ◽

Cited By ~ 1

Author(s):

Raoul L. Wientzen ◽

George H. McCracken ◽

Mary L. Petruska ◽

Susan G. Swinson ◽

Bertil Kaijser ◽

...

Keyword(s):

Urinary Tract ◽

Urinary Tract Infections ◽

Pediatric Patients ◽

Clinical Presentation ◽

C Reactive Protein ◽

E Coli ◽

Reactive Protein ◽

Tract Infections ◽

Antibody Coating

One hundred four patients with 124 episodes of urinary tract infection were studied. Serum C-reactive protein (CRP) was determined on diagnosis of each patient. Children with a CRP equal to or greater than 30 µg/ml (CRP-pos) differed significantly from those with values less than 30 µ/ml (CRP-neg) in age, clinical presentation, K type of Escherichia coli causing disease, frequency or radiographic abnormalities, and presence of antibody coating of bacteria in the urinary sediment. E coli K1 strains caused disease significantly more often in CRP-pos than in CRP-neg patients, and children with K1 infections were younger than those with non-K1 infections. The antibody-coated bacteria test was neither sensitive nor specific for localization of infection in pediatric patients. Determination of K1 antibody concentrations in serum and urine of E coli K1-infected children provided data supporting the measurement of CRP as one means of localizing urinary tract infections. Patients with CRP-neg infections were treated as successfully with four days of antimicrobial therapy as with ten days.

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A poly(ethylene glycol)-supported quaternary ammonium salt for highly efficient and environmentally friendly chemical fixation of CO2 with epoxides under supercritical conditions

Tetrahedron Letters ◽

10.1016/j.tetlet.2005.12.077 ◽

2006 ◽

Vol 47 (8) ◽

pp. 1271-1275 ◽

Cited By ~ 103

Author(s):

Ya Du ◽

Jin-Quan Wang ◽

Jian-Yu Chen ◽

Fei Cai ◽

Jie-Sheng Tian ◽

...

Keyword(s):

Ethylene Glycol ◽

Ammonium Salt ◽

Quaternary Ammonium ◽

Quaternary Ammonium Salt ◽

Environmentally Friendly ◽

Chemical Fixation ◽

Poly Ethylene Glycol ◽

Supercritical Conditions ◽

Highly Efficient ◽

Poly Ethylene

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Multifunctional periodontal membrane for treatment and regeneration purposes

Journal of Bioactive and Compatible Polymers ◽

10.1177/0883911520911659 ◽

2020 ◽

Vol 35 (2) ◽

pp. 117-138 ◽

Cited By ~ 1

Author(s):

Gulhan Isik ◽

Nesrin Hasirci ◽

Aysen Tezcaner ◽

Aysel Kiziltay

Keyword(s):

Stem Cells ◽

Vitamin E ◽

Ethylene Glycol ◽

Dental Pulp ◽

Drug Loading ◽

Dental Pulp Stem Cells ◽

Poly Ethylene Glycol ◽

E Coli ◽

Poly Ethylene

Periodontitis is a chronic inflammatory disease that causes gum tissue degeneration and alveolar bone and tooth loss. The aim of this study is to develop a multifunctional matrix for the treatment of periodontitis and enhancement of regeneration of the periodontal tissue. The matrix was prepared from vitamin E containing hydrogel made of alginate and gelatin, and doxycycline HCl containing methoxy poly(ethylene glycol)-block-polycaprolactone micelles. Methoxy poly(ethylene glycol)-block-polycaprolactone was synthesized with ring-opening polymerization technique and characterized by proton nuclear magnetic resonance (1H NMR), Fourier-transform infrared spectroscopy, differential scanning calorimetry, and gel permeation chromatography. Micelles were characterized by measuring zeta potential, hydrodynamic diameter, drug encapsulation efficiency, drug loading capacity, and in vitro drug-release kinetics. Micelles were obtained with an average size of 164 nm and drug loading amount of 5.8%. The activity of doxycycline HCl–loaded micelles and vitamin E containing hydrogels was determined against Escherichia coli ( E. coli) and Staphylococcus aureus ( S. aureus) with disk diffusion method. Bio-efficacy of micelle-loaded alginate–gelatin hydrogels were tested in vitro using L929 fibroblasts and dental pulp stem cells. Doxycycline HCl–loaded micelles and vitamin E containing hydrogels showed a sustained release and exhibited inhibition zone against E. coli and S. aureus. Hydrogels with vitamin E and doxycycline HCl–loaded micelles promoted osteogenic differentiation of dental pulp stem cells. Results suggest that alginate–gelatin hydrogels containing doxycycline HCl–loaded micelles and vitamin E can be good candidates for the treatment of periodontitis and tissue regeneration.

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Surface-Attached Poly(oxanorbornene) Hydrogels with Antimicrobial and Protein-Repellent Moieties: The Quest for Simultaneous Dual Activity

Materials ◽

10.3390/ma11081411 ◽

2018 ◽

Vol 11 (8) ◽

pp. 1411 ◽

Cited By ~ 5

Author(s):

Monika Kurowska ◽

Vania Widyaya ◽

Ali Al-Ahmad ◽

Karen Lienkamp

Keyword(s):

Antimicrobial Activity ◽

Surface Charge Density ◽

Cell Envelope ◽

Adhesion Assay ◽

Poly Ethylene Glycol ◽

E Coli ◽

Polymer Hydrogels ◽

Air Interface ◽

Protein Adhesion ◽

Poly Ethylene

By copolymerizing an amphiphilic oxanorbornene monomer bearing N- tert-butyloxycarbonyl (Boc) protected cationic groups with an oxanorbornene-functionalized poly(ethylene glycol) (PEG) macromonomer, bifunctional comb copolymers were obtained. Varying the comonomer ratios led to copolymers with PEG contents between 5–25 mol %. These polymers were simultaneously surface-immobilized on benzophenone-bearing substrates and cross-linked with pentaerythritoltetrakis(3-mercaptopropionate). They were then immersed into HCl to remove the Boc groups. The thus obtained surface-attached polymer hydrogels (called SMAMP*-co-PEG) were simultaneously antimicrobial and protein-repellent. Physical characterization data showed that the substrates used were homogeneously covered with the SMAMP*-co-PEG polymer, and that the PEG moieties tended to segregate to the polymer–air interface. Thus, with increasing PEG content, the interface became increasingly hydrophilic and protein-repellent, as demonstrated by a protein adhesion assay. With 25 mol % PEG, near-quantitative protein-adhesion was observed. The antimicrobial activity of the SMAMP*-co-PEG polymers originates from the electrostatic interaction of the cationic groups with the negatively charged cell envelope of the bacteria. However, the SMAMP*-co-PEG surfaces were only fully active against E. coli, while their activity against S. aureus was already compromised by as little as 5 mol % (18.8 mass %) PEG. The long PEG chains seem to prevent the close interaction of bacteria with the surface, and also might reduce the surface charge density.

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A Regenerative Immunoaffinity Layer Based on the Outer Membrane of Z-Domains Autodisplaying E. coli for Immunoassays and Immunosensors

Sensors ◽

10.3390/s18114030 ◽

2018 ◽

Vol 18 (11) ◽

pp. 4030 ◽

Cited By ~ 1

Author(s):

Daseul Jeon ◽

Jae-Chul Pyun ◽

Joachim Jose ◽

Min Park

Keyword(s):

Limit Of Detection ◽

Gold Surface ◽

Outer Cell ◽

Detergent Concentration ◽

Orientation Control ◽

E Coli ◽

Spr Biosensor ◽

Antibody Orientation ◽

Outer Cell Membrane ◽

The Cost

Through orientation control of antibodies, Z-domains autodisplaying Escherichia coli outer cell membrane (OM) may be utilized to improve the sensitivity and limit of detection (LOD) of immunoassays and immunosensors. A regenerative immunoaffinity layer based on Z-domains autodisplaying E. coli OM was developed for the surface plasmon resonance (SPR) biosensor. Regeneration conditions for the Z-domains autodisplaying E. coli OM-based immunoassays and immunosensors were optimized by varying pH and detergent concentration. An E. coli cell-based HRP immunoassay was tested and validated in three sequential regenerative immunoassays under optimal conditions. The OM of Z-domains autodisplaying E. coli was isolated and coated on the two-dimensional substrate (microplate). The OM-based HRP immunoassay was tested and validated in four regenerative immunoassays. This regenerative OM layer was applied to the SPR biosensor. Z-domains autodisplaying OM layered onto the gold surface of SPR biosensors was developed, and the OM-based regenerative immunoaffinity layer with orientation control was tested using CRP analyte. The SPR biosensor regenerative immunoaffinity layer demonstrated that CRP biosensing was repeated for five regeneration cycles with less than 2% signal difference. Therefore, the newly developed regenerative immunoaffinity layer with antibody orientation control may improve biosensing sensitivity and reduce the cost of medical diagnosis.

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