以纤维素滤纸膜为载体,染料Cibacron Blue F3GA为配基,制备了一种新型亲和膜色谱介质。采用扫描电镜、红外光谱、元素分析等方法对亲和膜介质进行鉴定与表征,该膜具有良好的色谱性能。亲和膜对F3GA的键合质量摩尔浓度达93.7μmol/g。研...以纤维素滤纸膜为载体,染料Cibacron Blue F3GA为配基,制备了一种新型亲和膜色谱介质。采用扫描电镜、红外光谱、元素分析等方法对亲和膜介质进行鉴定与表征,该膜具有良好的色谱性能。亲和膜对F3GA的键合质量摩尔浓度达93.7μmol/g。研究了木瓜蛋白酶在亲和膜上的吸附行为,实验表明:在30℃下、酶质量浓度为2 mg/mL、pH=8.0时,吸附质量比可达57.9 mg/g,改变pH值及离子强度等条件对吸附质量比有明显的影响。在最适条件下吸附遵循Langmuir型吸附。可以初步推断,纤维素滤纸膜可以制成性能优良的亲和膜色谱介质,成本低廉,适合工业化分离纯化生物大分子。展开更多
Bilirubin removal from human plasma was obtained via an affinity microporous polytetrafluoroethylene（MPTFE） capillary. The new adsorbent comprised grafted glycidyl methacrylate（GMA） via radiation-induced polymeriz...Bilirubin removal from human plasma was obtained via an affinity microporous polytetrafluoroethylene（MPTFE） capillary. The new adsorbent comprised grafted glycidyl methacrylate（GMA） via radiation-induced polymerization as hydrophilic coating and reactive sites; ethylenediamine（EDA） as a spacer arm; Cibacron Blue F3GA（CB F3GA） as an affinity ligand; MPTFE capillary as the supporting matrix. The average density of CB F3GA attachment to MPTFE capillaries was found to be 136.5 μmol/g. The capacity of bilirubin adsorbed on affinity MPTFE capillaries is 76.1 mg bilirubin/g polymer（at 25℃）. This new adsorbent has advantages over both membrane and traditional micro-column, and this system is stable and easy to operate. The results of blood tests suggest the CB F3GA affinity capillary has good blood compatibility.展开更多
Rigid PSDVB microbeads have been modified with poly(vinyl alcohol) (PVA) adsorbed on their surface to produce an affinity medium. Then Cibacron Blue F3GA was covalently attached to the supports. The initial concentrat...Rigid PSDVB microbeads have been modified with poly(vinyl alcohol) (PVA) adsorbed on their surface to produce an affinity medium. Then Cibacron Blue F3GA was covalently attached to the supports. The initial concentration of PVA has effect on the adsorption of PVA. The non-specific interaction of bovine serum albumin (BSA) on the microbeads decreases with the PVA adsorbed. The pH stability test shows that the affinity medium is stable up to pH 11.0. And it has specific interaction with lysozyme, but not with pepsin.展开更多
Urine is a potential source of diagnostic biomarkers for detection of diseases,and is a very attractive means of non-invasive biospecimen collection.Nonetheless,proteomic measurement in urine is very challenging becau...Urine is a potential source of diagnostic biomarkers for detection of diseases,and is a very attractive means of non-invasive biospecimen collection.Nonetheless,proteomic measurement in urine is very challenging because diagnostic biomarkers exist in very low concentration(usually below the sensitivity of common immunoassays)and may be subject to rapid degradation.Hydrogel nanoparticles functionalized with Cibacron Blue F3G-A(CB)have been applied to address these challenges for urine biomarker measurement.We chose one of the most difficult low abundance,but medically relevant,hormones in the urine:human growth hormone(hGH).The normal range of hGH in serum is 1 to 10 ng/mL but the urine concentration is suspected to be a thousand times less,well below the detection limit(50 pg/mL)of sensitive clinical hGH immunoassays.We demonstrate that CB particles can capture,preserve and concentrate hGH in urine at physiological salt and urea concentrations,so that hGH can be measured in the linear range of a clinical immunometric assay.Recombinant and cadaveric hGH were captured from synthetic and human urine,concentrated and measured with an Immulite chemiluminescent immunoassay.Values of hGH less than 0.05 ng/mL(the Immulite detection limit)were concentrated to 2 ng/mL,with a urine volume of 1 mL.Dose response studies using 10 mL of urine demonstrated that the concentration of hGH in the particle eluate was linearly dependent on the concentration of hGH in the starting solution,and that all hGH was removed from solution.Thus if the starting urine volume is 100 mL,the detection limit will be 0.1 pg/mL.Urine from a healthy donor whose serum hGH concentration was 1.34 ng/mL was studied in order detect endogenous hGH.Starting from a volume of 33 mL,the particle eluate had an hGH concentration of 58 pg/mL,giving an estimated initial concentration of hGH in urine of 0.175 pg/mL.The nanotechnology described here appears to have the desired precision,accuracy and sensitivity to support large scale clinical studies of urine hGH levels.展开更多
A series of aminated chitosan microspheres were prepared from chitosan,and then an affinity dye-ligand,Cibacron Blue F3GA,was covalently coupled.The maximum attachment of cibacron blue F3GA was 494 μmol/g.Albumin ads...A series of aminated chitosan microspheres were prepared from chitosan,and then an affinity dye-ligand,Cibacron Blue F3GA,was covalently coupled.The maximum attachment of cibacron blue F3GA was 494 μmol/g.Albumin adsorption onto cibacron blue F3GA-attached chitosan microspheres was investigated.The human serum albumin (HSA) equilibrium adsorption capacity was 79.8 mg/g (wet beads).The adsorption equilibrium isotherms appeared to follow a typical Langmuir isotherm.The highest desorption ratio (100%) was achieved by using an eluent of 1.0 mol/L NaSCN (pH 8.0).The cibacron blue F3GA-attached chitosan microspheres could be reused without significant decreases in the adsorption capacities.展开更多
以尼龙膜为基质,键合壳聚糖的方法对尼龙膜进行改性,使膜的非特异性吸附大大降低,并偶联活性染料亲和基Cibacron Blue F3GA,得到一种新的染料亲和膜.该亲和膜具有良好的色谱性能,对木瓜蛋白酶有较高的吸附量(235.3 mg/g),吸附行为满足Fr...以尼龙膜为基质,键合壳聚糖的方法对尼龙膜进行改性,使膜的非特异性吸附大大降低,并偶联活性染料亲和基Cibacron Blue F3GA,得到一种新的染料亲和膜.该亲和膜具有良好的色谱性能,对木瓜蛋白酶有较高的吸附量(235.3 mg/g),吸附行为满足Freundlich吸附模型,pH值和离子强度对吸附量有明显影响.使用该亲和膜对木瓜粉中的木瓜蛋白酶进行分离纯化,纯化倍数可达46.5倍.展开更多
An affinity adsorbent, Cibacron Blue 3GA immobilized magnetic polyvinyl alcohol (PVA) microspheres was used for bilirubin removal taking the advantage of easy separation of magnetic sorbent from the biosystem. Fe3O4 s...An affinity adsorbent, Cibacron Blue 3GA immobilized magnetic polyvinyl alcohol (PVA) microspheres was used for bilirubin removal taking the advantage of easy separation of magnetic sorbent from the biosystem. Fe3O4 superparamagnetic particles was synthesized with hydrothermal reaction of ferrous chloride (FeCl2) and ferric chloride (FeCl3). Such magnetic particles are then encapsulated in biocompatible PVA to form magnetic polymer microspheres sized from 2 to 15 nm with hydroxyl groups on its surface. Cibacron Blue 3GA, a dye-ligand, was covalently coupled with the polyvinyl alcohol through the nucleophilic reaction between the chloride of its triazine ring and the hydroxyl groups of PVA molecules under alkaline condition. The affinity adsorbent carried 21.1μmol Cibacron Blue 3GA per gram magnetic polymer microspheres was used to remove unconjugated and conjugated bilirubin from the solution which was composed of bilirubin or bilirubin and protein. After the adsorption, the adsorbent loaded with bilirubin was removed easily in the magnetic field.展开更多
Affinity dye-ligand Cibacron Blue F3GA（CB F3GA） was covalently coupled with poly（vinyl alcohol）（PVA） coated on the inner surface of microporous poly（tetra-fluoroethylene）（MPTFE） membranous capillary. The PVA...Affinity dye-ligand Cibacron Blue F3GA（CB F3GA） was covalently coupled with poly（vinyl alcohol）（PVA） coated on the inner surface of microporous poly（tetra-fluoroethylene）（MPTFE） membranous capillary. The PVA-coated PTFE capillary surface was characterized by XPS and FESEM. The grafting degree of PVA and the amount of CB F3GA immobilized onto the membranous capillary were 23.5 mg/g and 89.6 pmol/g, respectively. These dyed membranous capillaries were chemically and mechanically stable, and could be reproducibly prepared. Human serum albumin（HSA） was selected as model protein. The saturation adsorbance of the dye attached membranous capillary was 85.3 mg HSA/g, while the capacity of non-specific adsorption for HSA was less than 0.3 mg/g.展开更多
文摘以纤维素滤纸膜为载体,染料Cibacron Blue F3GA为配基,制备了一种新型亲和膜色谱介质。采用扫描电镜、红外光谱、元素分析等方法对亲和膜介质进行鉴定与表征,该膜具有良好的色谱性能。亲和膜对F3GA的键合质量摩尔浓度达93.7μmol/g。研究了木瓜蛋白酶在亲和膜上的吸附行为,实验表明:在30℃下、酶质量浓度为2 mg/mL、pH=8.0时,吸附质量比可达57.9 mg/g,改变pH值及离子强度等条件对吸附质量比有明显的影响。在最适条件下吸附遵循Langmuir型吸附。可以初步推断,纤维素滤纸膜可以制成性能优良的亲和膜色谱介质,成本低廉,适合工业化分离纯化生物大分子。
基金Supported by the National Natural Science Foundation of China(No.29405038)
文摘Bilirubin removal from human plasma was obtained via an affinity microporous polytetrafluoroethylene（MPTFE） capillary. The new adsorbent comprised grafted glycidyl methacrylate（GMA） via radiation-induced polymerization as hydrophilic coating and reactive sites; ethylenediamine（EDA） as a spacer arm; Cibacron Blue F3GA（CB F3GA） as an affinity ligand; MPTFE capillary as the supporting matrix. The average density of CB F3GA attachment to MPTFE capillaries was found to be 136.5 μmol/g. The capacity of bilirubin adsorbed on affinity MPTFE capillaries is 76.1 mg bilirubin/g polymer（at 25℃）. This new adsorbent has advantages over both membrane and traditional micro-column, and this system is stable and easy to operate. The results of blood tests suggest the CB F3GA affinity capillary has good blood compatibility.
文摘Rigid PSDVB microbeads have been modified with poly(vinyl alcohol) (PVA) adsorbed on their surface to produce an affinity medium. Then Cibacron Blue F3GA was covalently attached to the supports. The initial concentration of PVA has effect on the adsorption of PVA. The non-specific interaction of bovine serum albumin (BSA) on the microbeads decreases with the PVA adsorbed. The pH stability test shows that the affinity medium is stable up to pH 11.0. And it has specific interaction with lysozyme, but not with pepsin.
基金This work was partly supported by the Italian Istituto Superiore di Sanita’in the framework of the Italy/USA cooperation agreement between the U.S.Department of Health and Human Services,George Mason University and the Italian Ministry of Public HealthThis work was partially supported by the U.S.Department of Energy grant number DE-FC52-04NA25455。
文摘Urine is a potential source of diagnostic biomarkers for detection of diseases,and is a very attractive means of non-invasive biospecimen collection.Nonetheless,proteomic measurement in urine is very challenging because diagnostic biomarkers exist in very low concentration(usually below the sensitivity of common immunoassays)and may be subject to rapid degradation.Hydrogel nanoparticles functionalized with Cibacron Blue F3G-A(CB)have been applied to address these challenges for urine biomarker measurement.We chose one of the most difficult low abundance,but medically relevant,hormones in the urine:human growth hormone(hGH).The normal range of hGH in serum is 1 to 10 ng/mL but the urine concentration is suspected to be a thousand times less,well below the detection limit(50 pg/mL)of sensitive clinical hGH immunoassays.We demonstrate that CB particles can capture,preserve and concentrate hGH in urine at physiological salt and urea concentrations,so that hGH can be measured in the linear range of a clinical immunometric assay.Recombinant and cadaveric hGH were captured from synthetic and human urine,concentrated and measured with an Immulite chemiluminescent immunoassay.Values of hGH less than 0.05 ng/mL(the Immulite detection limit)were concentrated to 2 ng/mL,with a urine volume of 1 mL.Dose response studies using 10 mL of urine demonstrated that the concentration of hGH in the particle eluate was linearly dependent on the concentration of hGH in the starting solution,and that all hGH was removed from solution.Thus if the starting urine volume is 100 mL,the detection limit will be 0.1 pg/mL.Urine from a healthy donor whose serum hGH concentration was 1.34 ng/mL was studied in order detect endogenous hGH.Starting from a volume of 33 mL,the particle eluate had an hGH concentration of 58 pg/mL,giving an estimated initial concentration of hGH in urine of 0.175 pg/mL.The nanotechnology described here appears to have the desired precision,accuracy and sensitivity to support large scale clinical studies of urine hGH levels.
文摘A series of aminated chitosan microspheres were prepared from chitosan,and then an affinity dye-ligand,Cibacron Blue F3GA,was covalently coupled.The maximum attachment of cibacron blue F3GA was 494 μmol/g.Albumin adsorption onto cibacron blue F3GA-attached chitosan microspheres was investigated.The human serum albumin (HSA) equilibrium adsorption capacity was 79.8 mg/g (wet beads).The adsorption equilibrium isotherms appeared to follow a typical Langmuir isotherm.The highest desorption ratio (100%) was achieved by using an eluent of 1.0 mol/L NaSCN (pH 8.0).The cibacron blue F3GA-attached chitosan microspheres could be reused without significant decreases in the adsorption capacities.
文摘以尼龙膜为基质,键合壳聚糖的方法对尼龙膜进行改性,使膜的非特异性吸附大大降低,并偶联活性染料亲和基Cibacron Blue F3GA,得到一种新的染料亲和膜.该亲和膜具有良好的色谱性能,对木瓜蛋白酶有较高的吸附量(235.3 mg/g),吸附行为满足Freundlich吸附模型,pH值和离子强度对吸附量有明显影响.使用该亲和膜对木瓜粉中的木瓜蛋白酶进行分离纯化,纯化倍数可达46.5倍.
基金Supported by the National Natural Science Foundation of China (No. 29776036).
文摘An affinity adsorbent, Cibacron Blue 3GA immobilized magnetic polyvinyl alcohol (PVA) microspheres was used for bilirubin removal taking the advantage of easy separation of magnetic sorbent from the biosystem. Fe3O4 superparamagnetic particles was synthesized with hydrothermal reaction of ferrous chloride (FeCl2) and ferric chloride (FeCl3). Such magnetic particles are then encapsulated in biocompatible PVA to form magnetic polymer microspheres sized from 2 to 15 nm with hydroxyl groups on its surface. Cibacron Blue 3GA, a dye-ligand, was covalently coupled with the polyvinyl alcohol through the nucleophilic reaction between the chloride of its triazine ring and the hydroxyl groups of PVA molecules under alkaline condition. The affinity adsorbent carried 21.1μmol Cibacron Blue 3GA per gram magnetic polymer microspheres was used to remove unconjugated and conjugated bilirubin from the solution which was composed of bilirubin or bilirubin and protein. After the adsorption, the adsorbent loaded with bilirubin was removed easily in the magnetic field.
基金Supported by the National Natural Science Foundation of China(No29405038)
文摘Affinity dye-ligand Cibacron Blue F3GA（CB F3GA） was covalently coupled with poly（vinyl alcohol）（PVA） coated on the inner surface of microporous poly（tetra-fluoroethylene）（MPTFE） membranous capillary. The PVA-coated PTFE capillary surface was characterized by XPS and FESEM. The grafting degree of PVA and the amount of CB F3GA immobilized onto the membranous capillary were 23.5 mg/g and 89.6 pmol/g, respectively. These dyed membranous capillaries were chemically and mechanically stable, and could be reproducibly prepared. Human serum albumin（HSA） was selected as model protein. The saturation adsorbance of the dye attached membranous capillary was 85.3 mg HSA/g, while the capacity of non-specific adsorption for HSA was less than 0.3 mg/g.
