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ABSTRACT: A rapid, effective method for the screening of adsorbent ligands based on the unique optical properties of gold colloid has been developed. Different interactions between adsorbate and ligands induce different states of aggregation of the gold colloid, and the associated distinct color changes of the colloid have been utilized to estimate the affinity of the ligands toward the adsorbate. In this work, phosphorylated peptide CGGFGGpSG was appended to a gold colloid to obtain the adsorbate-modified gold colloid (CG8-AuNPs). Candidate ligands Dpa-Zn(2+), DMAPAA, and AAn were copolymerized with acrylamide to form linear polymers and cross-linked CG8-AuNPs to induce aggregation. Screening of the candidate ligands revealed that Dpa-Zn(2+) showed the highest affinity among those tested, inducing a color change of the gold colloid at a concentration of 10 μM, which is much lower than those of ligands DMAPAA (40 μM) and AAn (almost no color change could be observed). Subsequent statistical adsorption experiments confirmed these screening results, with the adsorbent A-AAm-Dpa-Zn(2+) showing the highest adsorption capacity (426 mg/g) for CG-8, almost twice that of adsorbent A-AAm-DMAPAA. This reported method has low sample consumption, and the screening may be simply monitored by the naked eye.
Langmuir 08/2011; 27(18):11457-65. · 4.19 Impact Factor
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Jun Wang,
Saihui Zhang,
Jing Zhang,
Ping Ren,
Yingchao Chen,
Jihong Li,
Wei Wang,
Yi Ma,
Rongfu Shi,
Chunhong Wang,
Zhi Yuan
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ABSTRACT: In this study, with flavonol glycosides (FG) and terpene lactones (TL) in ginkgo biloba extract (GBE) as the targets for separation, we investigated the effectiveness of molecular docking in adsorbent screening. Several polyamine-modified methyl acylate-co-divinylbenzene (MA-co-DVB) adsorbent models were built, and their affinity to rutin, quercetin and ginkgolide B (GB) was evaluated via molecular docking. The model of ethylenediamine-modified adsorbent showed the largest difference in affinity between to GB and to quercetin as well as rutin, and thus this adsorbent could have the best separation performance. The results of the subsequently conducted static adsorption and dynamic adsorption experiments correlated well with docking results. Finally, using ethylenediamine-modified MA-co-DVB adsorbent, nearly complete separation of the FG and TL in GBE was simply achieved by one step of adsorption-desorption. Thus, the reported molecular docking method is expected to be helpful for rapid adsorbent screening.
Journal of chromatography. B, Analytical technologies in the biomedical and life sciences 06/2011; 879(19):1605-9. · 2.78 Impact Factor
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ABSTRACT: To investigate the role of the peptide Pro-His-Ser-Arg-Asn (PHSRN) in cell adhesion and growth, PHSRN- and Gly-Arg-Gly-Asp-Ser (GRGDS)-containing polymers (P-PN5 and P-GS5, respectively) were synthesized by modification of poly(D,L-lactide-co-beta-malic acid) (PLMA) with the corresponding peptides. The cell affinities of the modified polymers were evaluated by adhesion and proliferation experiments with human umbilical vein endothelial cells (HUVECs). The results showed that P-PN5 had comparable ability to that of P-GS5 in supporting HUVEC adhesion and growth. Furthermore, the integrin-mediated mechanism of cell-substrate interaction was investigated. The results showed that P-PN5 had similar binding affinity and binding strength towards α(5)β(1) compared to those of P-GS5. The findings suggest that PHSRN is capable of mediating the adhesion and growth of HUVECs independently and that PHSRN-modified polymers might be used as biologically compatible materials.
Colloids and surfaces. B, Biointerfaces 05/2011; 84(1):6-12. · 2.60 Impact Factor
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ABSTRACT: This work aimed to compare two types of affinity ligands, i.e. polymeric and monomeric ligands, by investigating their adsorption affinity, capacity and selectivity to oligopeptide. The peptide NH(2)-VVRGCTWW-COOH (VW-8) was chosen as the target adsorbate, while histidine (His), aspartic acid (Asp), and leucine (Leu) were selected as the ligands, respectively. For each kind of ligand, both monomeric (M) and polymeric (P) forms were introduced onto the Sepharose matrix respectively to obtain the corresponding adsorbents. Both affinity tests using isothermal titration calorimetry (ITC) and adsorption capacities using static adsorption experiments indicated that the adsorbents with polymeric ligands (MX-P) exhibited better adsorption ability for VW-8 than the adsorbents with monomeric ligands (MX-M). In particular, the MX-PHis exhibited its affinity constant of 2.39 × 10(6) M(-1) and its adsorption capacity of 77.4 mg/g for VW-8, which was approximately 8-10 times higher than that of MX-MHis. Such distinct adsorption abilities between polymeric and monomeric ligands were interpreted based on nuclear magnetic resonance (NMR) and ITC data, and the results indicated that such better characters of polymeric ligands were ascribed to their good flexibility which facilitated the cooperative effects as well as the accessibility of ligands to the peptide. Additionally, the selective adsorption experiments indicated that all the adsorbents with polymeric ligands exhibited good selectivity to the peptide VW-8.
Journal of Colloid and Interface Science 03/2011; 355(2):478-85. · 3.07 Impact Factor
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ABSTRACT: A major challenge in the development of affinity adsorbents is the design of specific adsorbents for target molecules. In this paper, a two-step strategy was used to design a specific adsorbent for oligopeptides. Based on the structural characteristics of target peptide DFLAE (DE5), the affinity ligand CDenHis bearing hydrophobic inclusion and electrostatic interaction sites was prepared by grafting histidine onto β-cyclodextrin (CD) using ethylenediamine; ligands with single hydrophobic inclusion or electrostatic interaction sites (CDen and HisOMe) were used as reference ligands. Results indicated that the binding affinity (K(a)) of CDenHis with DE5 was 6.23×10(4)M(-1), 23- and 61-fold higher than that of CDen and HisOMe, respectively. Computer simulations were used to further optimize the steric configuration of CDenHis. It was found that the optimized ligand CDdnHis exhibited a much improved binding affinity for DE5 (K(a)=1.02×10(5)M(-1)). Moreover, the corresponding adsorbent A-CDdnHis not only showed much better adsorption ability compared with A-CDenHis, but also excellent adsorption specificity for DE5-containing peptides. Kinetic analysis and adsorption mechanism studies suggested that the configuration matching of CDdnHis with DE5 and the cooperation of multiple interactions led to the fast and selective adsorption of DE5-containing peptides to A-CDdnHis.
Journal of chromatography. A 10/2010; 1217(48):7539-46. · 4.19 Impact Factor
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ABSTRACT: Asp-Phe-Leu-Ala-Glu (DE5) is a frequent sequence of many toxic middle molecular peptides that accumulate in uremic patients. To eliminate these peptides by hemoperfusion, three adsorbents (CP1-Zn(2+), CP2-Zn(2+), and CP3-Zn(2+)) were designed on the basis of coordination and hydrophobic interactions. Adsorption experiments indicated that CP2-Zn(2+) had the highest affinity for DE5 among these three adsorbents. Also, the adsorption capacity of CP2-Zn(2+) in DE5 and DE5-containing peptides was about 2-6 times higher than that of peptides without the DE5 sequence. Linear polymers bearing the same functional groups of the adsorbents were used as models to study the adsorption mechanism via isothermal titration calorimetry (ITC) and computer-aided analyses. The results indicated that coordination and hydrophobic interactions played the most important roles in their affinity. When two carboxyl moieties on Asp and Glu residues coordinated to CP2-Zn(2+), the hydrophobic interaction took place by the aggregation of the hydrophobic amino acid residues with phenyl group on CP2-Zn(2+). The optimal collaboration of these interactions led to the tight binding and selective adsorption of DE5-containing peptides onto CP2-Zn(2+). These results may provide new insight into the design of affinity adsorbents for peptides containing DE5-like sequences.
Langmuir 03/2010; 26(10):7181-7. · 4.19 Impact Factor
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ABSTRACT: Endothelialization is an ideal approach to improve the blood compatibility of synthetic polymers. However, cell detachment is inevitable under shear flow conditions. Therefore, the issue of blood compatibility needs to be addressed for both the bare and the endothelialized polymer. RGD-containing polymer P-GS5 was synthesized by modification of poly(D,L-lactide-co-beta-malic acid) (PLMA) with the peptide GRGDS. The compositions, molecular weights and hydrophilicities of poly(D,L-lactide) (PDLLA), PLMA, and P-GS5 were characterized by nuclear magnetic resonance (NMR), X-ray photoelectron spectroscopy (XPS), gel-permeation chromatography (GPC) and water contact angle measurements, respectively. The blood compatibilities of the bare and the endothelialized polymers were evaluated by clotting time and platelet adhesion tests. The results showed that the coagulation pathways were not influenced before and after cell culture; the bare P-GS5 attracted less platelet adhesion and induced lower pseudopodia extension compared with PDLLA and PLMA, and the platelet adhesion on P-GS5 was almost completely eliminated after cell seeding. The results suggest that P-GS5 could be a potentially useful material in vascular tissue engineering.
Colloids and surfaces. B, Biointerfaces 09/2009; 75(1):370-6. · 2.60 Impact Factor
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ABSTRACT: Despite their increasing uses for cardiovascular and cerebrovascular tissue engineering, synthetic polymeric conduits still have their limitations in clinical applications, particularly in small vessels, mainly due to the thrombus formation. Seeding the synthetic scaffolds with endothelial cells (ECs) will potentially solve this problem, but this endothelialization approach demands synthetic materials with better hemacompatibility and cell affinity. To improve the currently used materials and screen for better surface properties, we synthesized copolymer of poly(lactide-co-beta-malic acid) (PLMA), and its derivatives with pendant hydroxyl arms (PLMAHE) or extended carboxyl arms (PLMA-ECA). We analyzed their physical and chemical properties, their hydrophilicity, and their degradation in physiological conditions. More importantly, their blood compatibility was investigated by the measurements of prothrombin time, activated partial thromboplastin time, and interaction with platelets; their cell affinity and cell growth potentials were observed using the human umbilical vein EC cultures. Results from these experiments showed that the copolymer with the carboxyl arms attracted little platelets, and exhibited better cell affinity and supported the cell proliferation, thus demonstrating the potential usefulness of PLMA-ECA for tissue engineering. We speculate that this novel material will offer new opportunities for the design of better vascular-engineered scaffolds owing to its improved biological and chemical properties.
Tissue Engineering Part A 01/2009; 15(1):65-73. · 4.64 Impact Factor
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ABSTRACT: Protein phosphorylation plays an important role in intracellular signaling. Defects in the phosphorylation process, which are associated with cancers and other human diseases, can be reflected by the presence of abnormally phosphorylated peptides in serum. Therefore, phosphorylated peptides are important resources for biomarker discovery. However, few studies have been reported on developing sensors for phosphorylated peptides. In this study, zinc(ii) dipicolylamine-appended gold nanoparticles were synthesized and used for the colorimetric detection of bis-phosphorylated peptides. The addition of the bis-phosphorylated peptides could induce the aggregation of the functionalized AuNPs, and thus the color change of the gold colloid, while non-phosphorylated and mono-phosphorylated controls could not induce the color change.
Sensors and Actuators B: Chemical.
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ABSTRACT: In order to raise the yield of hydrocortisone (HC) from cortexolone-21-acetate (RSA), 11 β-hydroxylase-producing Absidia orchidis was entrapped in calcium alginate gel and the bioconversion was conducted in cosolvent-containing media. The optimal immobilization procedure for Absidia orchidis was determined. The factors of influence on the conversion were studied in detail. The optimal concentrations of the cosolvent, detergent, and substrate in the reaction system were investigated. The optimal pH and temperature of the biotransformation were also determined. It was found that the ratio of HC(11 β-isomer) and 11 α-hydroxyl hydrocortisone (α-HC) in the products was 2.2–2.5 and the yield of HC reached 59–61% in a cosolvent-containing system. This can be compared with the ratio of HC and α-HC(HC/α-HC) of 0.9 and HC yield of 40% in an aqueous system. Stability studies showed that the immobilized cells can be used for six times without any significant decrease in activity. The results reported in this paper show a high potential in industrial production of HC.
Enzyme and Microbial Technology 22(5):368-373. · 2.37 Impact Factor
