Dissolved oxygen and O-2 flux across the water-air interface of the Changjiang Estuary in May 2003

The field observation of this study was carried out in the Changjiang Estuary from May 19 to 26,2003, just a few days before the Three Gorges Dam began to store water. A total of 29 stations, including 2 anchor stations, were distributed through almost the whole salinity gradient Based on the data gained from these stations, the biogeochemical characteristics of dissolved oxygen (DO) were examined. Spatial distribution of DO concentrations showed the pattern that it increased in a downriver direction. DO concentration generally varied within a narrow range of 733-8.10 mg l(-1) in the freshwater region and the west part of the mixed water region, and after that it increased rapidly. In vertical direction, the differences in DO concentrations between surface and 2 m above the bottom were big at the stations with water depths exceeding 20 m; DO concentration up to 14.88 mg l(-1) was recorded at the sea surface, while at 2 m above the bottom its concentration was only about 4 mg l(-1). The fluctuation in DO concentrations was small during a period of 48 h in the mixed water region and 2 m above the bottom of the seawater region; while it was large during the same period in the seawater region for surface and 5 m below the surface layer, and a maximum variation from 8.77 to 12.66 mg l(-1) in 4 h was recorded. Oxygen fluxes also showed a marked spatio-temporal variation. As a whole, the freshwater region and mixed water region were an oxygen sink while the seawater region was a source. Relationships between dissolved oxygen and some biogeochemical parameters which could markedly influence its spatio-temporal distribution were discussed in this paper. (C) 2008 Elsevier B.V. All rights reserved.

Phage M13Ko7 Detection With Biosensor Based On Imaging Ellipsometry And Afm Microscopic Confirmation

A rapid detection and identification of pathogens is important for minimizing transfer and spread of disease. A label-free and multiplex biosensor based on imaging ellipsometry (BIE) had been developed for the detection of phage M13KO7. The surface of silicon wafer is modified with aldehyde, and proteins can be patterned homogeneously and simultaneously on the surface of silicon wafer in an array format by a microfluidic system. Avidin is immobilized on the surface for biotin-anti-M13 immobilization by means of interaction between avidin and biotin, which will serve as ligand against phage M13KO7. Phages M13KO7 are specifically captured by the ligand when phage M13KO7 solution passes over the surface, resulting in a significant increase of mass surface concentration of the anti-M13 binding phage M13KO7 layer, which could be detected by imaging ellipsometry with a sensitivity of 10(9) pfu/ml. Moreover, atomic force microscopy is also used to confirm the fact that phage M13KO7 has been directly captured by ligands on the surface. It indicates that BIE is competent for direct detection of phage M13KO7 and has potential in the field of virus detection. (C) 2008 Elsevier B.V. All rights reserved.

果实成熟衰老与抗病性的调控机制研究

　　果实为开花植物所特有的发育器官，在种子的成熟和传播过程中发挥着重要作用。同时，肉质果实中含有丰富的营养物质，包括纤维素、维生素、抗氧化剂等，成为人们饮食的重要组成部分。由于果实的成熟衰老和抗病性直接影响果品的质量和市场价值，因此，研究果实成熟衰老和抗病性的调控机制具有重要的理论意义和应用前景。本文主要利用蛋白质组学的方法，探讨外源化学物质抑制果实成熟衰老和诱导抗病性的调控机制。 1. 硅对果实的抗病性诱导：用硅酸钠（1%）处理采后的甜樱桃果实，再接种褐腐病原菌（Molinilia fracticola），置于20C下，观测贮藏期间果实的发病率，并分析硅处理后诱导的主要蛋白质及调控机制。研究结果表明：硅酸钠处理可显著抑制贮藏期间褐腐病的发生，其抑病机理与硅诱导PR-蛋白的表达，提高果实的抗氧化水平，减轻由病原菌侵染造成的氧化胁迫相关。同时，硅处理还能保护细胞骨架结构，有利于增强果实对病原菌入侵的抵抗力。 2. 水杨酸对果实的抗病性诱导：用水杨酸（SA，2mM）在果园处理三种成熟度的甜樱桃果实，然后接种青霉病原菌（Penicillium expansum）观察其发病情况，并取样分析参与抗病性应答的主要蛋白质及调控机制。试验结果表明：SA处理能显著降低青霉病的发病率和抑制病斑扩展，而且SA对低成熟度甜樱桃果实的抗性诱导效果更好。在八成熟的果实中，有5个热激蛋白和4个脱氢酶蛋白被SA诱导，这些蛋白参与了糖酵解和三羧酸循环。抗氧化蛋白和PR蛋白主要参与较低成熟度果实的抗性应答，而热激蛋白和脱氢酶在较高成熟度果实的抗性应答中更明显，SA诱导的抗性与代谢途径相关。 　　3. 草酸对果实的抗性诱导：用5mM的草酸处理冬枣果实后，接种青霉菌（P. expansum），观察果实发病情况，测定果实相关的生理指标，分析参与果实抗性应答的主要蛋白质及调控机制。结果表明：草酸能明显延缓冬枣果实的衰老，提高果实对青霉菌的抗性。草酸处理能抑制果实乙烯的释放量和呼吸强度，延缓叶绿素的降解，减少乙醇积累。利用蛋白质组学的研究方法证实了在25个参与了草酸处理应答的蛋白中，胱硫醚-β-合酶结构域包含蛋白（CBB domain-containing protein）和3个与光合作用相关蛋白[二磷酸核酮糖羧化酶/加氧酶（Ribulose bisphosphate carboxylase/oxygenase activase, chloroplast precursor），二磷酸核酮糖羧化酶/加氧酶大亚基结合蛋白（RuBisCO large subunit-binding protein subunit beta, chloroplast precursor），植物光系统Ⅱ放氧复合蛋白2（PSII oxygen-evolving complex protein 2）]的表达量上调，乙醇脱氢酶的表达量出现下调。草酸处理还提高了与乙烯合成前体相关蛋白的表达，抑制了ACC合成酶的活性。草酸提高果实抗病的机制与延缓果实成熟衰老和保持果实抗性有关。 4. 果实衰老的调控机制：采用高氧（100%）和低氧（2-3%）处理苹果果实，观察果实衰老的进程，并基于蛋白质组学的研究方法，探讨苹果果实衰老与线粒体蛋白质组的关系。结果表明，在苹果衰老过程中有22个蛋白的表达量发生变化，这些蛋白主要参与了三羧酸循环，电子传递，碳代谢和胁迫应答。高氧处理能诱导氧化胁迫，加速了果实的衰老。质谱鉴定结果证明：在高氧胁迫下，超氧化物歧化酶（manganese superoxide dismutase，MnSOD）和线粒体外膜通道蛋白（porin) 的表达量降低，MnSOD的活性受到抑制，由此提高了线粒体中超氧阴离子的含量，增加了蛋白质的氧化损伤。 此外，高氧处理改变了porin的功能，导致了线粒体膜的透势发生变化，从而引起外膜损伤。由此阐明了活性氧在果实的成熟衰老调控中的重要作用。

mRNA 很可能携带三维遗传信息

The forming mechanism of the three - dimensional structures of proteins，i.e.the mechanism of protein folding，is a basic problem in molecular biology which is still unsolved unitl now. In which a core problem is whether there is the three – dimensional genetic information that decide the three - dimensional structures of proteins. However, the research on this field has mot yet been reported. Recently，we made a comparative study on the folded structures of more than 70 mature messeneger RNAs (mRNAs) and the three - dimensional structures of the proteins encoded by them，it has been found that there exist marked correspondences between their featured structures in the following aspects: 1.The number of the structural units. An RNA molecule can form a secondary structure(stem and loop structure) by the folding and the base pairing of itself. The elementary structural unit of an RNA secondary structure is hairpin(or compound hair pin).The regular structural unit in the secondary structure of a protein is # alpha # - helix or #beta# - sheet . We have found that the hairpin number in the secondary structure of each mature mRNA is equal or approximately equal to the number of the regular secondary structural unis of the encoded protein. 2 .Turning region. Turn is a main structrual element in the secondary structure of a protein, which decides the backbone orientation of a protein molecule to some extent .Our analysis shows that the nucleotide sequence segments in an mRNA which encode the turns of the corresponding protein are overall situated in the turning regions of the mRNA secondary structure such as haipin，bulge loop or multibaranch loops. 3 .The arrangement of structural elements in space. In order to understand the backbone orientation of an RNA molecule and the arangement of its structural elements in space，we have modeled the three一dimensional structure of the mRNA molecule on SGI workstation based on its secondary structure.The result shows that the spatial arrangement of most of the nucleotide sequence segments encoding the structural elements of a protein is consistent with that of these stretural exements in the protein. For instance，the nucleotide sequences corresponding to each pleated sheet of a # beta # - sheet structure are close to each other in the mRNA secondary stucture and in the three - dimensional structure，although some of the nucleotide segments are far apart from each other in the one - dimensional sequence. For another instance，the two triplet codons of cysteines which form a disulphide bridge geneal1y are very close to each other in the mRNA folded structure. In addition，we also analyzed the locations of the codons proline - coding and the distrbution of the nucleotide sequences #alpha# - helix - coding in the folded structures of mRNAs . Some distribution laws have been found. All of these results suggest that the transfer of the genetic information from mRNA to protein not only is one – dimensional but also is three - dime ns ional. That is，there exists the genetic information that decide the three - dimensional structures of proteins. To a certain extent，we could say that the mRNA folding detemines the protein folding. Based on these results，it would be possible to predict the three - dimensional structures of proteins from the primary，secondary and tertiary structures of the m RNAs at a higher accuracy.And more important is that a new clue has been provided to uncover the“spatial coding" of the genetic information.

Identification of differentially expressed genes in Tetrahmena thermophila in response to dichlorodiphenyltrichloroethane (DDT) by suppression subtractive hybridization

The insecticide dichlorodiphenyltrichloroethane (DDT) is persistent in the environment, and continues to cause health problems. Tetrahymena has potential as a model organism for assaying low levels of DDT and for analysing the mechanisms of its toxicity. We constructed the suppression subtractive hybridization library of T thermophila exposed to DDT, and screened out 90 Expressed Sequence Tags whose expressions were significantly up- or downregulated with DDT treatment. From this, a series of important genes related to the DDT metabolism and detoxification were discovered, such as P450 gene, glutathione S-transferase gene and sterol carrier protein 2 gene. Furthermore, their expressions under different concentrations of DDT treatment were detected by real-time fluorescent quantitative PCR. The results show that Tetrahymena is a relevant and useful model organism for detecting DDT in the environment and for discovering biomarkers that can be used to develop specific bio-reporters at the molecular and genomic levels.

Shape-gradient composite surfaces: Water droplets move uphill

The approach of water droplets self-running horizontally and uphill without any other forces was proposed by patterning the shape-gradient hydrophilic material (i.e., mica) to the hydrophobic matrix (i.e., wax or low-density polyethylene (LDPE)). The shape-gradient composite surface is the best one to drive water droplet self-running both at the high velocity and the maximal distance among four different geometrical mica/wax composite surfaces. The driving force for the water droplets self-running includes: (1) the great difference in wettability of surface materials, (2) the low contact angle hysteresis of surface materials, and (3) the space limitation of the shape-gradient transportation area. Furthermore, the average velocity and the maximal distance of the self-running were mainly determined by the gradient angle (alpha), the droplet volume, and the difference of the contact angle hysteresis. Theoretical analysis is in agreement with the experimental results.

Is inorganic nutrient enrichment a driving force for the formation of red tides? A case study of the dinoflagellate Scrippsiella trochoidea in an embayment

Red tides (high biomass phytoplankton blooms) have frequently occurred in Hong Kong waters, but most red tides occurred in waters which are not very eutrophic. For example, Port Shelter, a semi-enclosed bay in the northeast of Hong Kong, is one of hot spots for red tides. Concentrations of ambient inorganic nutrients (e.g. N, P), are not high enough to form the high biomass of chlorophyll a (chl a) in a red tide when chl a is converted to its particulate organic nutrient (N) (which should equal the inorganic nutrient, N). When a red tide of the dinoflagellate Scrippsiella trochoidea occurred in the bay, we found that the red tide patch along the shore had a high cell density of 15,000 cells ml(-1), and high chl a (56 mu g l(-1)), and pH reached 8.6 at the surface (8.2 at the bottom), indicating active photosynthesis in situ. Ambient inorganic nutrients (NO3, PO4, SiO4, and NH4) were all low in the waters and deep waters surrounding the red tide patch, suggesting that the nutrients were not high enough to support the high chl a >50 mu g l(-1) in the red tide. Nutrient addition experiments showed that the addition of all of the inorganic nutrients to a non-red-tide water sample containing low concentrations of Scrippsiella trochoidea did not produce cell density of Scrippsiella trochoidea as high as in the red tide patch, suggesting that nutrients were not an initializing factor for this red tide. During the incubation of the red tide water sample without any nutrient addition, the phytoplankton biomass decreased gradually over 9 days. However, with a N addition, the phytoplankton biomass increased steadily until day 7, which suggested that nitrogen addition was able to sustain the high biomass of the red tide for a week with and without nutrients. In contrast, the red tide in the bay disappeared on the sampling day when the wind direction changed. These results indicated that initiation, maintenance and disappearance of the dinoflagellate Scrippsiella trochoidea red tide in the bay were not directly driven by changes in nutrients. Therefore, how nutrients are linked to the formation of red tides in coastal waters need to be further examined, particularly in relation to dissolved organic nutrients. (C) 2008 Elsevier B.V. All rights reserved.

东喜马拉雅构造结新生代碰撞变形与岩石圈演化

The Eastern Himalayan Syntaxis (EHS) is one of the strongest deformation area along the Himalayan belt resulted from the collision between Indian plate and the Eurasian Plate since the 50～60Ma, and has sensitivity tracked and preserved the whole collisional processes. It should depend on the detail geological investigations to establish the deformational accommodate mode, and the uplift history, to elucidate the deep structure and the crust-mantle interaction of the Tibet Plateau of the EHS. The deep-seated (Main Mantle Thrusts) structures were exhumed in the EHS. The MMT juxtapose the Gangdese metamorphic basement and some relic of Gangdese mantle on the high Himalayan crystalline series. The Namjagbawa group which is 1200～1500Ma dated by U/Pb age of zircon and the Namla group which is 550Ma dated by U/Pb age of zircon is belong to High Himalayan crystalline series and Gangdese basement respectively. There is some ophiolitic relic along the MMT, such as metamorphic ocean mantle peridotite and metamorphic tholeiite of the upper part of ocean-crust. The metamorphic ocean mantle peridotites (spinel-orthopyroxene peridotite) show U type REE patterns. The ~(87)Sr/~(86)Sr ratios were, 0.709314～0.720788, and the ~(143)Nd/~(144)Nd ratios were 0.512073～0.512395, plotting in the forth quadrant on the ~(87)Sr/~(86)Sr-~(143)Nd/~(144)Nd isotope diagram. Some metamorphic basalt (garnet amphibolite) enclosures have been found in the HP garnet-kynite granulite. The garnet amphibolites can be divided two groups, the first group is deplete of LREE, and the second group is flat or rich LREE, and their ~(87)Sr/~(86)Sr, ~(143)Nd/~(144)Nd ratios were 0.70563～0.705381 and 0.512468～0.51263 respectively. Trace element and isotopic characteristics of the garnet amphibolites display that they formed in the E-MORB environment. Some phlogolite amphibole harzburgites, which exhibit extensive replacement by Phl, Amp, Tc and Dol etc, were exhumed along the MMT. The Phl-Amp harzburgites are rich in LREE and LILE, such as Rb, K etc, and depletes Eu (Eu~* = 0.36 ～ 0.68) and HFSE, such as Nb, Ta, Zr, Hf, P, Ti etc. The trace element indicate that the Phl-Amp harzburgites have island arc signature. Their ~(87)Sr/~(86)Sr are varied from 0.708912 to 0.879839, ~(143)Nd/~(144)Nd from 0.511993 to 0.512164, ε Nd from- 9.2 to - 12.6. Rb/Sr isochrone age of the phlogolite amphibole harzburgite shows the metasomatism took place at 41Ma, and the Amp ~(40)Ar/~(39)Ar cooling age indcate the Phl-Amp harzburgite raising at 16Ma. There is an intense crust shortening resulted from the thrust faults and folds in the Cayu block which is shortened more 120km than that of the Lasha block in 35～90Ma. With the NE corner of the India plate squash into the Gangdese arc, the sinistral Pai shear fault and the dextral Aniqiao shear fault on the both sides of the Great bent of Yalun Zangbu river come into active in 21～26Ma. On the other hand, the right-lateral Gongrigabu strike-slip faults come into activity at the same period, a lower age bound for the Gongrigabu strike-slip fault is estimated to be 23～24Ma from zircon of ion-probe U/Pb thermochronology. The Gongrigabu strike-slip faults connect with the Lhari strike-slip fault in the northwestern direction and with the Saganing strike-slip at the southeastern direction. Another important structure in the EHS is the Gangdese detachment fault system (GDS) which occurs between the sedimental cover and the metamorphic basement. The lower age of the GDS is to be 16Ma from the preliminary 40Ar/39Ar thermochronology of white mica. The GDS is thought to be related to the reverse of the subducted Indian crust and the fast uplift of the EHS. Structural and thermochronology investigation of the EHS suggest that the eastern Tibet and the western Yunnan rotated clockwise around the EHS in the period of 35～60Ma. Later, the large-scale strike-slip faults (RRD, Gaoligong and Saganing fault) prolongate into the EHS, and connect with the Guyu fault and Gongrigabu fault, which suggest that the Indianchia block escape along these faults. Two kind of magmatic rocks in the EHS have been investigated, one is the mantle-derived amphibole gabbro, dioposide diorite and amphibole diorite, another is crust origin biotit-garnet adamellite, biotit-garnet granodiorite and garnet-amphibole-biotite granite. The amphibole gabbro dioposite diorite and amphibole diorite are rich in LREE, and LILE, such as Ba, Rb, Th, K, Sr etc, depleted in HFSE, such as Nb, Ta, Zr, Hf, Ti etc. The ratio of ~(87)Sr/~(86)Sr are from 0.7044 to 0.7048, ~(143)Nd/~(144)Nd are from 0.5126 to 0.5127. The age of the mantle origin magamatic rocks, which result from the partial melt of the raising and decompression anthenosphere, is 8Ma by ~(40)Ar/~(39)Ar dating of amphibole from the diorite. The later crust origin biotite-garnet adamellite, biotite-garnet granodiorite and garnet-amphibole-biotite granite are characterized by aboudance in LREE, and strong depletion of Eu. The ratios of ~(87)Sr-~(86)Sr are from 0.795035 to 0.812028, ~(143)Nd/~(144)Nd from 0.51187 to 0.511901. The ~(40)Ar/~(39)Ar plateau age of the amphibole from the garnet-amphibole-biotite granite is 17.5±0.3Ma, and the isochrone age is 16.8±0.6Ma. Their geochemical characteristics show that the crust-derived magmatic rocks formed from partial melting of the lower curst in the post-collisional environment. A group of high-pressure kaynite-garnet granulites and enclave of high-pressure garnet-clinopyroxene grnulites and calc-silicate grnulites are outcroped along the MMT. The peak metamorphic condition of the high-pressure granulites yields T=800～960 ℃, P=1.4～1.8Gpa, corresponding the condition of 60km depth. The retrograde assemblages of the high-pressure grnulites occur at the condition of T=772.3～803.3 ℃, P=0.63～0.64Gpa. The age of the peak metamorphic assemblages are 45 ～ 69Ma indicated by the zircon U/Pb ion-plobe thermochronology, and the retrograde assemblage ages are 13～26Ma by U/Pb, ~(40)Ar/~(39)Ar thermochronology. The ITD paths of the high-pressure granulites show that they were generated during the tectonic thickening and more rapid tectonic exhumation caused by the subducting of the Indian plate and subsequent break-off of the subducted slab. A great deal of apatite, zircon and sphene fission-track ages, isotopic thermochronology of the rocks in the EHS show that its rapid raising processes of the EHS can be divided into three main periods. There are 35～60Ma, 13～25Ma, 0～3Ma. 3Ma is a turn in the course of raising in the EHS which is characterized by abruptly acceleration of uplifting. The uplift ratios are lower than 1mm .a~(-1) before 3Ma, and higher than 1mm .a~(-1) with a maximum ratio of 30mm .a~(-1) since 3Ma. The bottom (knick point) of the partial anneal belt is 3.8km above sea level in the EHS, and correspond to age of 3Ma determined by fission-track age of apatite. The average uplift ratio is about 1.4 mm .a~(-1) below the knick point. The EHS has raised 4.3km from the surface of 2.36km above sea level since 3Ma estimated by the fossil partial anneal belt of the EHS. We propose a two-stage subduction model (B+A model) basing on Structural, thermochronological, magmatical, metamorphic and geophysical investigations of the EHS. The first stage is the subduction of the Indian continental margin following after the subduction of the Tethys Ocean crust and subsequent collision with the Gangdese arc, and the second stage is the Indian crust injecting into the lower crust and upper mantle of the Tibet plateau. Slab break-off seems to be occurred between these two stages.

Soluble Angiopoietin Receptor Tie-2 In Patients With Acute Myocardial Infarction And Its Detection By Optical Protein-Chip

The Tie-2 receptor has been shown to play a role in angiogenesis in atherosclerosis. The conventional method assaying the level of soluble Tie-2 (sTie-2) was ELISA. However, this method has some disadvantages. The aims of this research are to establish a more simple detection method, the optical protein-chip based on imaging ellipsomtry (OPC-IE) applying to Tie-2 assay. The sTie-2 biosensor surface on silicon wafer was prepared first, and then serum levels of sTie-2 in 38 patients with AMI were measured on admission (day 1), day 2, day 3 and day 7 after onset of chest pain and 41 healthy controls by ELISA and OPC-IE in parallel. Median level of sTie-2 increased significantly in the AMI patients when compared with the controls. Statistics showed there was a significant correlation in sTie-2 results between the two methods (r=0.923, P0.01). The result of this study showed that the level of sTie-2 increased in AMI, and OPC-IE assay was a fast, reliable, and convenient technique to measure sTie-2 in serum.

Nanoparticle-amplified Surface Plasmon Resonance Study of Protein Conformational Change at Interface

This paper reports the study of protein conformational change by Au nanoparticles (AUNPs)-amplified surface plasmon resonance (SPR) spectroscopy. Taking cytochrome c (Cyt c) as an example, this paper gives a detailed description of the construction of metal-protein-metal sandwich nanostructure consisting of an Au film underlayer, a cytochrome c intermediate layer and an AuNPs upper layer. The incorporation of AuNPs into SPR biosensing results in increased SPR sensitivity to protein conformational change as demonstrated by acid denaturation of Cyt c. It suggests the conformational change of surface-confined Cyt c leads to the distance and electromagnetic coupling variations of Au film-AuNPs.

Feasibility of Protein a for the Oriented Immobilization of Immunoglobulin on Silicon Surface for a Biosensor with Imaging Ellipsometry

The feasibility of using protein A to immobilize antibody on silicon surface for a biosensor with imaging ellipsometry was presented in this study. The amount of human IgG bound with anti-IgG immobilized by the protein A on silicon surface was much more than that bound with anti-IgG immobilized by physical adsorption. The result indicated that the protein A could be used to immobilize antibody molecules in a highly oriented manner and maintain antibody molecular functional configuration on the silicon surface. High reproducibility of the amount of antibody immobilization and homogenous antibody adsorption layer on surfaces could be obtained by this immobilization method. Imaging ellipsometry has been proven to be a fast and reliable detection method and sensitive enough to detect small changes in a molecular monolayer level. The combination of imaging ellipsometry and surface modification with protein A has the potential to be further developed into an efficient immunoassay protein chip.

Application of Optical Protein-chip in Detecting Phage M13KO7

Abstract: Avidin layer was bound on the substrate surface of Silicon wafer modified with aldehyde. The interaction between avidin and biotin was adopted for the immobilization of mouse monoclonal biotin-anti-M13 (antibody GP3)-labeled biotin. The surface was incubated in a solution containing phage M13KO7, which was trapped by the antibody GP3 with the interaction between phage M13KO7 and antibody GP3, resulting in a variation of layer thickness that was detected by imaging ellipsometry. The results showed a saturated layer of antibody GP3 with a thickness about 6.9 nm on the surface of the silicon wafer. The specific interaction between phage M13KO7 and antibody GP3 resulted in a variation of layer thickness. The layer of phage M13KO7 bound with antibody GP3 was 17.5 nm in the concentration of 1.1×1010 pfu/mL. Each variation of the layer thickness corresponded to a concentration of phage M13KO7 in the range of 0.1×1010–2.5×1010 pfu/mL, with the sensitivity of 109 pfu/mL. Compared with other methods, the optical protein-chip, requiring only short measurement time, label free, is a quantitative test, and can be visualized. This study could be significant on the interactions between the antibody and the virus, showing potential in the early diagnosis of virosis.

Measuring Binding Kinetics Of Surface-Bound Molecules Using The Surface Plasmon Resonance Technique

Surface plasmon resonance (SPR) technology and the Biacore biosensor have been widely used to measure the kinetics of biomolecular interactions in the fluid phase. In the past decade, the assay was further extended to measure reaction kinetics when two counterpart molecules are anchored on apposed surfaces. However, the cell binding kinetics has not been well quantified. Here we report development of a cellular kinetic model, combined with experimental procedures for cell binding kinetic measurements, to predict kinetic rates per cell. Human red blood cells coated with bovine serum albumin and anti-BSA monoclonal antibodies (mAbs) immobilized on the chip were used to conduct the measurements. Sensor-grams for BSA-coated RBC binding onto and debinding from the anti-BSA mAb-immobilized chip were obtained using a commercial Biacore 3000 biosensor, and analyzed with the cellular kinetic model developed. Not only did the model fit the data well, but it also predicted cellular on and off-rates as well as binding affinities from curve fitting. The dependence of flow duration, flow rate, and site density of BSA on binding kinetics was tested systematically, which further validated the feasibility and reliability of the new approach. Crown copyright (c) 2008 Published by Elsevier Inc. All rights reserved.