溴酚蓝测定血清白蛋白质的研究

用酸性染料为探针对蛋白质的定量已应用于临床医学及生命科学的研究中, 本文利用分光光度法研究溴酚蓝(BPB)与牛血清白蛋白(BSA)、人血清白蛋白(HSA)的反应. 在pH=2.79的Britton-Robinson缓冲溶液中, 血清白蛋白与溴酚蓝通过分子间力形成蓝色复合物使吸光度增加. λ=605 nm时, εBSA=5.20×105 L·(mol·cm)-1, εHSA=5.12×105 L·(mol·cm)-1;BSA在0～78.0 mg·L-1, HSA在0～81.6 mg·L-1范围内服从比尔定律. 检出限分别为BSA: 2.93 mg*L-1, HSA:3.12 mg·L-1. 对7个人血清蛋白总量平行6次测定, 相对标准偏差1.2%～3.1%, 回收率91.9%～105.6%.

Effect of Pore-size of Mesoporous SBA-15 on Adsorption of Bovine Serum Albumin and Lysozyme Protein

In this paper,the effects of pore-size of SBA-15 on the adsorption kinetics and equilibrium of large protein molecules Bovine serum albumin(BSA)and lysozyme(LYS)have been investigated.The mesoporous molecular sieve SBA-15 with six different pore sizes were synthesized with P123 triblock copolymer as the template agent,and 1,3,5-trimethylbenzene(TMB)and isopropyl alcohol as the pore-expanding agent.The samples were characterized by N2 adsorption/desorption,Scanning Electron Microscopy(SEM),Transmission Electron Microscopy(TEM)and X-Ray Diffraction(XRD).It is found that BSA and LYS were adsorbed rapidly on SBA-15 materials with large pores.The BSA adsorption capacity of sieve with the pore diameter of 21.4 nm reached 500 mg·g-1 within 25 minutes.However,if the pore diameter was smaller than 14 nm,the BSA adsorption capacity of the sieve was only about 220 mg·g-1.The adsorption equilibrium data fits in the Langmuir model,where the coefficient of effective use of specific area of mesoporous molecular sieve was found to be 0.03,0.18,0.37 and 0.48,corresponding to the pore diameter of 10.1 nm,13.2 nm,15.4 nm and 21.4 nm,respectively.The equilibrium loading amount of LYS on SBA-15 materials with pore size of 15.4 nm could be up to 1000 mg·g-1.The coefficient of effective use of surface area of mesoporous molecular sieve with diameter of 3.9 nm,7.4 nm,10.1 nm,13.2 nm and 15.4 nm was 0.10,0.47,0.56,0.71 and 0.79,respectively.It is also noted that greater pore size of mesoporous molecular sieve would lead to a higher coefficient of effective use of surface area.

Diagnostic application of serum proteomic patterns in early gastric cancer patients by SELDI-TOF-MS

Objective:Surface-enhanced laser desorption/ionization time-of-flight mass spectrometry(SELDI-TOF-MS) is one of the currently used techniques to identify biomarkers for cancers.This study was planned to make a proteomic analysis on the serum of stage I gastric cancer patients and establish a early diagnostic model for identifying stage I gastric cancer preliminarily.Methods:A total of 229 serum samples including 169 pathologically confirmed gastric cancer patients(stage I:n = 47;stage II:n = 39;stage III:n = 46;stage IV:n = 37) and 60 gastritis patients were analyzed by SELDI-TOF-MS Protein Chip array technology.The resulting SELDI-TOF-MS spectral data were analyzed using the Biomarker WizardTM and Biomarker Patterns software to find differential proteins and develop a classification tree for early gastric cancer.Results:Twelve proteins were significantly over expressed(P<0.05,Student t-test) in patients with gastric cancer compared to patients with gastritis.Among them,three protein masses with 2873,6121 and 7778 m/z was found to be differentially expressed solely in patients suffering from stage I gastric cancer.The three protein masses construct the decision tree which could effectively distinguish stage I gastric cancer samples from control samples,achieving a sensitivity and specificity of 89.36(42/47) and 97.32%(182/187),respectively.Conclusion:Serum Proteomic Patterns can be used for the quick diagnosis of stage I gastric cancer.SELDI-TOF-MS is a useful tool for the detection and identification of new protein markers in serum.

A highly sensitive “turn-on” fluorescent probe for bovine serum albumin protein detection and quantification based on AIE-active distyrylanthracene derivative

A sulfonated 9,10-distyrylanthracene derivative with aggregation-induced emission (AIE) property is designed and synthesized. It shows a highly sensitive and selective fluorescence enhancement property for bovine serum albumin (BSA) protein detection and quantification. Analysis on the interaction between the probe molecule and BSA reveals the essential role of the hydrophobic cavities of the protein folding structure.

A novel organic-inorganic hybrid monolith for trypsin immobilization

In proteomics, attention has focused on various immobilized enzyme reactors (IMERs) for the realization of high throughput digestion. In this report, a novel organic-inorganic hybrid monolith based IMER was prepared in a 100 μm i.d. capillary with 3-glycidoxypropyltrimethoxysilane (GLYMO) as the monomer and tetraethoxysilane (TEOS) as the crosslinker. Trypsin immobilization was achieved via the reaction between vicinal diol groups, which were obtained from hydrolysis of epoxy groups, and the amino groups of trypsin. Bovine serum albumin was digested thoroughly by this IMER in 47 s. After micro-reverse phase liquid chromatography-tandem mass spectrometry (μRPLC-MS/MS) analysis and database searching, beyond 35% sequence coverage was obtained, and the result was comparable to that of 12 h in solution digestion. The present IMER has potential for high throughput digestion.