Capillary electrophoresis with electrochemiluminescence detection of procyclidine in human urine pretreated by ion-exchange cartridge

This paper describe a Ru(bpy)(3)(2+) based electrochemiluminescence (ECL) method to detect procyclidine in human urine following separation by capillary electrophoresis (CE). An ECL detection cell was designed for post-column addition of Ru(bpy)(3)(2+). Parameters affecting separation and detection were optimized, leading to a detection limit of 1 x 10(-9) mol/l in an on-capillary stacking mode. For application in urine, a cartridge packed with slightly acidic cation-exchange resin was used to eliminate the matrix effects of urine and improve the detection sensitivity. Extraction recovery was nearly 90%.

Computer simulation of insoluble Pr(III) speciation in human interstitial fluid

A multi-phase model of Pr(III) speciation in human interstitial fluid was constructed and insoluble Pr(III) speciation was studied. When the total concentration of Pr(III) is below 8.401E-10 mol/L, soluble Pr(III) species are main species. With rising the total concentration of Pr(III), Pr(III) is firstly bound to phosphate to form precipitate of PrPO4, then bound to carbonate and another precipitate of Pr-2(CO3)(3) was obtained. When the total concentration is between 1.583E-9 mol/L and 4.000E-3 mol/L, the insoluble species are predominant Pr(III) species.

Computer simulation of human iodothyronine deiodinase single-chain antibody

Iodothyronine plays a major role in growth, basic metabolism and organ formation. It has an extremely limited source in the body. In this thesis, we designed iodothyronine(T4) as hapten. Then a single chain antibody displayed on phange was obtained from a human phage displaying a single chain antibody library. The specific genes of E3 was subcloned in P-5E vector. According to its amino acid sequences, we simulate its three dimention structure by computer. It has never been reported in PDB.

Amplified immunoassay of human IgG using real-time biomolecular interaction analysis (BIA) technology

An automated biomolecular interaction analysis instrument (BI-Acore) based on surface plasmon resonance (SPR) has been used to determine human immunoglobulin G (IgG) in real time. Polyclonal anti-human IgG antibody was covalently immobilized to a carboxymethyldextran modified gold film surface. The samples of human IgG prepared in HBS buffer were poured over the immobilized surface. The signal amplification antibody was applied to amplify the response signal. After each measurement, the surface was regenerated with 0.1 mol/L H3PO4. The assay was rapid, requiring only 30 min for antibody immobilization and 20 min for each subsequent process of immune binding, antibody amplification and regeneration. The antibody immobilized surface had good response to human IgG in the range of 0.12-60 nmol/L with a detection limit of 60 pmol/L. The same antibody immobilized surface could be used for more than 110 cycles of binding, amplification and regeneration. The results demonstrate that the sensitivity, specificity and reproducibility of amplified immunoassay using real-time BIA technology are satisfactory.

Detection of anti-human serum albumin antibody using surface plasmon resonance biosensor

The biosensor based on surface plasmon resonance(SPR) technology is a very useful tool to study the interaction between biomolecles. The main advantages of this technique is to "visualize" macromolecular interactions directly in real time, and in a label-free mode rather than indirect methods like enzyme-linked immunosorbent assays (ELISAs). We immobilize human serum albumin (HSA) to the carboxymethyldextran-modified sensor chip surface covalently to detect the activity of anti-HSA in serum, and regenerate the surface with .1 mol/L phosphoric acid. The results show that SPR biosensor can detect the activity of anti-HSA in real-time quickly and the sensor chip can be used over 100 cycles.

Determination of ultra-trace rare-earth elements in human plasma by inductively coupled plasma mass spectrometry

Inductively coupled plasma mass spectrometry (ICP-MS),a highly sensitive inorgnic analytic technique,fits to determine ultra-nace rare-earth elements in human plasma. Under the optimized conditions detection limits for 15 rare-earth elements are in the range of 0.7 (for Eu)-5.4 (for Gd) ng.L-1. Indium as an internal standard element is used to compensate for matrix suppression effect and sensitivity drift. Three kinds of preparation methods, diluted with 1% HNO3, digested with HNO3-H2O2 and with HNO3-HClO4, are checked and compared,and the former is the simplest way to be measured. The samples diluted with 1% HNO3, stored in 4 degrees C, are very steady for 16 days. With the method, 11 healthy plasma samples in Changchun area of China are analysed.

Studies of soluble human catalytic antibody with glutathione peroxidase activity

By screening the phage-displayed human single chain antibody library, we have got the specific single chain antibody bound to GSH-S-DNP butyl ester as the hapten. The tertiary structure of the protein was analyzed with the aid of computer, and the results showed the CDR3 region located on the surface of the antibody. The soluble antibody was expressed in E. coli. and the active site serine was converted into selenocysteine with the chemical modifying method, which resulted in the catalytic antibody with GPx activity of 80 U/mu mol. Furthermore, the same Ping-Pong mechanism as the natural GPx was observed when the kinetic behavior of the antibody was studied.

Studies on insoluble species of Gd (III) in human blood plasma by computer simulation

The insoluble species of Gd (III) in human blood plasma were investigated by computer simulation. The distribution of the Gd (I) species was obtained. It was found that most of the Gd (III) ions were bound to phosphate to form precipitate GdPO4 at the concentration of 1. 000 x 10(-7) mol/L and when the concentration of the Gd (III) increased to 3. 750 x 10(-4) mol/L, in excess of the concentration of phosphate, the Gd (III) ions were bound to carbonate to form another kind of precipitate, Gd-2 (CO3)(3).

Tb(III) speciation in human blood plasma by computer simulation

A multi-phase model was developed and Tb(III) speciation in human blood plasma was studied. At a concentration below 3.744x 10(-4) mol/L (or at the concentration), Tb(III) is mostly bound to phosphate to form precipitate of TbPO4. As the concentration of Tb(III) increases, phosphate is exceeded and another kind of precipitate of Tb-2(CO3)(3) appears. Among soluble Tb(III) species, Tb(III) mainly distribute in [Tb (Tf)] at low concentration and in [Tb (HSAA, [Tb-2 (Tf)], [Th (IgG)], [Tb (Lactate)](2+), [Tb (CitArgH)] and free Tb(III) at high concentration.

Human catalytic antibodies with glutathione peroxidase activity

In order to generate catalytic antibodies with glutathione peroxidase (GPx) activity, we prepared GSH-S-DNP butyl ester and GSH-S-DNP benzyl ester as the haptens. Two ScFvs that bound specifically to the haptens were selected from the human phage-displayed antibody library. The two ScFv genes were highly homologous, consisting of 786 bps and belonging to the same VH family-DP25. In the premise of maintaining the amino acid sequence, mutated plasmids were constructed by use of the mutated primers in PCR, and they were over-expressed in E. coli. After the active site serine was converted into selenocysteine with the chemical modifying method, we obtained two human catalytic antibodies with GPx activity of 72.2U/mu mol and 28.8U/mu mol, respectively. With the aid of computer mimicking, it can be assumed that the antibodies can form dimers and the mutated selenocysteine residue is located in the binding site. Furthermore, the same Ping-Pong mechanism as the natural GPx was observed when the kinetic behavior of the antibody with the higher activity was studied. (C) 2001 Elsevier Science BY. All rights reserved.

Studies on the transport of lanthanides ions through the membrane of human erythrocyte in vitro by ICP-MS

A method was developed for the determination of lanthanum in the cytoplasm of human erythrocytes after they were incubated in lanthanum nitrate or citrate solutions. The lanthanum concentration in the cytoplasm of incubated erythrocytes is much higher than that in normal erythrocytes. It is suggested lanthanum can transport through the membrane of erythrocyte in vitro. Solutions containing chelator are unsuitable to be washing buffer in the investigation.

Biological effect of rare earth (I) - Content and distribution of rare earth in normal human plasma

The content and distribution of rare earth(RE) in normal human plasma have been investigated by ultrafiltration, FPLC and ICP-MS methods, The results showed that there are trace RE in normal human plasma, and their contents are in accordance with their abundance, The RE can bond with immunoglobulin G(IgG), transferrin(Tf) and albumin(Alb) species, but mostly bond with Tf.

NMR studies of Gd(DTPA-BIN) in human serum albumin solution

Stable gadolinium complexes, such as Gd(DTPA) and Gd(DOTA), are usually used as the contrast agents for magnetic resonance imaging(MRI). Reported here are the enhanced relaxation properties of a novel gadolinium complex, diethylene-triaminopentaacetate Lis (isoniazid) [Gd(DTPA-BIN)], in aqueous and in human serum albumin(HSA) solution, which indicates that (1) two Gd(DTPA-BIN) can integrate non-covalently with one HSA with an equilibrium constant of 0. 02 mmol(-2) . L-2 ; (2) the relaxivities are 3. 28 and 4. 92 mmol(-1) . L . s(-1) for the free Gd(DTPA-BIN) and the [Gd(DTPA-BIN)](2), HSA conjugator, respectively; (3) the rotational correlation time of protein conjugator is notably higher than that of the free complex, The above results may imply that Gd(DTPA-BIN) has a higher tissue selectivity than that of its parent Gd(DTPA).

INTERACTION OF LANTHANUM AND CALCIUM WITH HUMAN ERYTHROCYTE-MEMBRANES

The effect of lanthanum and calcium on the structure and function of human erythrocyte membranes was investigated by fluorescence polarization, spin- labeled electron spin resonance (ESR) and laser Raman spectroscopy. The results showed that low concentration of La3+ (0.5 mu mol/L) activated a Little (Na++K+)-ATPase and Mg2+-ATPase activities, and it inhibited obvi ously the ATPase activities with increasing its concentrations. La3+ lowered the lipid fluidity of human erythrocyte membranes and decreased the vibration intensity of alpha-helix of the protein in the Amide I '. The effect of Ca2+ on the lipid fluidity and alpha-helix of the protein in the Amide I ' was smaller than that of La3+.

MULTIVARIATE CLASSIFICATION BASED ON METAL CONTENTS IN HUMAN SENILE CATARACT LENSES

Multivariate classification methods were used to evaluate data on the concentrations of eight metals in human senile lenses measured by atomic absorption spectrometry. Principal components analysis and hierarchical clustering separated senile cataract lenses, nuclei from cataract lenses, and normal lenses into three classes on the basis of the eight elements. Stepwise discriminant analysis was applied to give discriminant functions with five selected variables. Results provided by the linear learning machine method were also satisfactory; the k-nearest neighbour method was less useful.