妊娠相关血浆蛋白A定量测定方法的建立

目的:建立人外周血妊娠相关血浆蛋白A(PAPP-A)定量酶联免疫测定方法.方法:以生物素化牛血清白蛋白-链霉亲和素间接包被微孔板,将标记有生物素的抗PAPP-A抗体作为捕获抗体,结合标本中的抗原成分;标记有辣根过氧化物酶的抗PAPP-A抗体作为检测抗体,经酶催化底物显色,定量测定人体外周血PAPP-A.结果:所建立的测定方法灵敏度为0.31 mg/L,回收率均在95%～100%之间;该方法线性、稳定性及特异性良好;3份低、中、高浓度的血清标本重复测定10次,批内变异系数(CV)均小于10%,批间CV均小于15%;该方法测量结果与国外同类方法测量结果具有相关性(r=0.982,P<0.05),2种检测方法高度相关.结论:本文采用间接包被技术所建立的人外周血PAPPA固相酶联免疫测定方法灵敏、可靠、稳定性好,与国外同类方法的试剂盒之间相关性较高.     

疫苗中残留卵清蛋白ELISA定量测定方法的建立

目的 建立ELISA法检测疫苗中残留卵清蛋白的含量.   方法 将进口纯化兔抗卵清蛋白抗体作为包被抗体,辣根过氧化物酶标记兔抗卵清蛋白抗体作为酶标抗体,以系列含量的卵清蛋白溶液为标准,采用ELISA双抗体夹心法,对供试品中残留卵清蛋白进行定量测定.   结果 最佳线性范围为1.25～20 ng/ml,相关系数r≥0.99;定量限度为1.25 ng/ml;准确性试验回收率为89.2%～103.6%;精密性试验内和试验间变异系数分别为5.6%～6.7%和4.2%～9.4%.与马血清、羊血清、人血清白蛋白、牛血清白蛋白、流行性感冒病毒裂解疫苗基质液及其他无残留卵清蛋白的疫苗均无交叉反应.   结论 本方法特异性强、灵敏度高,准确性、重复性和稳定性好,可用于疫苗中残留卵清蛋白的定量检测.     

苦参碱白蛋白微球的制备及性质

以乳化-化学交联法制备苦参碱白蛋白微球。采用扫描电镜、差示扫描量热法和X-射线衍射法考察微球的特性,并用动态透析法考察体外释药规律。所得微球外观圆整,包封率为(83.55±3.17)%,载药量为(6.19±0.24)%,平均粒径(0.94±0.15)μm。体外释放试验结果显示,苦参碱白蛋白微球具有明显的缓释作用。     

间接ELISA测定血清中RI含量方法的建立

目的：建立检测血清中核糖核酸酶抑制因子（Ribonuclease Inhibitor，RI）含量的问：陵酶联免疫吸附法（EUSA），并分析该方法的特异性和敏感性。方法：用血清样品包被酶标板，兔抗人RI抗体为一抗，辣根过氧化物酶标记的羊抗兔抗体为二抗，建立血清RI定量检测的间接EUSA法。结果：所建立的用于血清中RI含量检测的间接ELISA方法特异性好．灵敏度高。结论：间接ELISA法可以用于测定血清中的RI抗原含量，具有良好的灵敏度和稳定性，能够有效的反映出人血清中的RI表达情况。     

抗癌药物白蛋白微球的研究进展

为提高抗癌药物对癌细胞或组织的靶向性,增强疗效,降低其全身毒副作用,以不同材料作载体的抗癌药物微球相继研制成功:不可生物降解的乙基纤维素微球,可生物降解的白蛋白微球、淀粉微球、明胶微球、聚乳酸微球,以及近年来问世的磁性微球、毫微球等。白蛋白微球以其良好的生物相容性和可降解性被广泛用于抗癌药靶向给药系统。 白蛋白微球最初用于动物及人的肺部扫描和循环系统研究,自1974年首次作为抗癌药物载体以来,相继用作诊断试剂及靶向给药、化学栓塞治疗的载体。1 抗癌药物动脉栓塞白蛋白微球 肿瘤动脉栓塞疗法是将抗癌药物制剂选择性注入支配     

斑蝥素白蛋白微球制备工艺的优化

目的优化斑蝥素白蛋白微球的制备工艺,并对其形态学性质、载药量进行考察.方法以加热固化法制备微球,以均匀设计考察影响因素.结果微球平均粒径为(0.91±0.16)μm,载药量为(15.32±1.15)%.结论优化后工艺较合理,所制得微球形态圆整、均匀,载药量较高.     

内分泌激素的磁分离酶联免疫技术检测

磁分离酶联免疫技术(MAIA)是20世纪80年代中期由瑞士Serono诊断中心发明的一种非同位素免疫检测的先进技术(美国专利号4659678)，亦称为磁性抗体免疫技术。这种技术是与免疫酶系统(磁性颗粒)结合，使用两种高亲和力单克隆抗体的一种测定法，它使双抗体夹心酶联免疫技术在对游离型标记抗体和抗原即抗体复合物分离中完全快速地分离。     

心脏靶向超声造影剂空气白蛋白微球的研究(1)

按正交设计筛选了用发射超声能量合成白蛋白空气微球的较佳制备工艺,并对微球的形态、粒径与粒径分布、稳定性、安全性和显影效果进行了系统研究。结果表明：微球的平均直径为 ４．２±０． ２ μｍ,平均浓度为（３．５±１．２）×１０８／ｍｌ。９２．５％的微球直径小于１０μｍ,微球在２℃～５℃条件下能稳定储存１２个月,经兔外用静脉注射后能顺利通过肺微循环到达左心室致使左心室靶向超声显影。     

核素标记叶酸偶联白蛋白纳米微球对人卵巢癌细胞生长的影响

目的观察核素标记叶酸靶向白蛋白纳米微球对人卵巢癌细胞生长的影响。方法将体外培养的SKOV3人卵巢癌细胞分为八组:阴性对照组(只加RPMI-1640培养液),单纯化疗组(叶酸偶联载药白蛋白磁性纳米微球,不加磁场),单纯放疗组[188铼(188 Re)标记的叶酸偶联白蛋白磁性纳米微球,不加磁场],单纯热疗组(叶酸偶联白蛋白磁性纳米微球,加磁场),化疗联合放疗组(188 Re标记的叶酸偶联载药白蛋白磁性纳米微球,不加磁场),化疗联合热疗组(叶酸偶联载药白蛋白磁性纳米微球,加磁场),放疗联合热疗组(188 Re标记的叶酸偶联白蛋白磁性纳米微球,加磁场)和热疗、化疗、放疗联合治疗组(联合治疗组,188 Re标记的叶酸偶联载药白蛋白磁性纳米微球,加磁场)。48h后MTT方法测定各组细胞增殖率,流式细胞术测定细胞凋亡率。结果阴性对照组对卵巢癌细胞增殖的抑制作用弱于其他各组,联合治疗组强于其他各组(P<0.05)。单纯热疗组、化疗联合放疗组、化疗联合热疗组、放疗联合热疗组和联合治疗组中细胞周期G1期前出现明显的亚二倍体凋亡峰;阴性对照组、单纯热疗组、化疗联合放疗组、化疗联合热疗组、放疗联合热疗组和联合治疗组的细胞凋亡率分别为0.08%、7.56%、17.14%、21.64%、33.94%和57.16%。结论磁感应热疗、化疗、核素靶向放疗的联合作用能有效抑制人卵巢癌细胞的生长。     

肿瘤动脉栓塞化疗用白蛋白微球的研究

用白蛋白制备了动脉栓塞用微球,该微球平均粒径为45±10μm,药物含量10.2%。通过渗透膜的体外释放试验中,7d 的总释药量为27%,而溶液剂6 h 的释药量达80%。动物实验表明,白蛋白微球可以栓塞末稍动脉。     

Differences in the serum protein binding of prazosin in man and rat

The serum protein binding of prazosin in man and rat has been studied in vitro by equilibrium dialysis. Prazosin was more extensively bound in human serum than in rat serum with binding ratios (B/F) of 14.3 +/- 3.4 and 4.4 +/- 0.2 (corresponding to 93.4 and 81.4% bound), respectively. This difference in binding between the species was partly due to qualitative differences between human and rat serum albumin, but also to the lower concentration of albumin in rat serum. Rat serum albumin (RSA) apparently showed two different classes of binding sites for prazosin, one with high (KD = 5.78 X 10(-6) M) and one with low (KD = 1.1 X 10(-4) M) affinity; the former is suggested as representing alpha 1-acid glycoprotein (alpha 1-AGP) with one binding site for prazosin per molecule, the latter as representing RSA with 0.28 binding sites per molecule. Human serum albumin (HSA) and human alpha 1-AGP both showed one class of binding sites with KD values of 2.7 X 10(-5) and 1.95 X 10(-6) M, respectively. HSA possessed 0.5 and human alpha 1-AGP 1 binding site for prazosin per molecule. The binding parameters obtained for the isolated serum proteins overestimated to some degree the total serum protein binding of prazosin in man. This was explained by a specific deviation from the law of mass action. HSA was the major binding protein in human serum at therapeutic concentrations, with ca. 60% of the total binding, the remaining 40% being bound to alpha 1-AGP. Anticipating that the high affinity binding site on the RSA preparation represents the binding of prazosin to alpha 1-AGP, then this protein accounts for 70% of the binding in rat serum, while rat serum albumin accounts for approximately 23%. The binding of prazosin to lipoproteins was insignificant in both species. The observed differences between man and rat in the serum protein binding of prazosin implicate differences in the two species with respect to prazosin pharmacokinetics and the pharmacological effect.     

Hydrolysis of carbaryl by human serum albumin

Human serum (HS) and human serum albumin (HSA) were able to hydrolyse the carbamate carbaryl. Carbarylase activity found in HSA was slightly activated by 1 mM Zn²⁺, Mn²⁺, Cd²⁺, Ni²⁺ and Na⁺ and by 0.01 mM Pb²⁺. The organophosphorus compounds paraoxon and O-hexyl O-2,5-dichlorophenyl phosphoramidate, caprylic acid, palmitic acid and the carboxyl ester p-nitrophenyl butyrate inhibited the hydrolysis of carbaryl by HSA, being in the last case a competitive inhibition. Using selective amino acid reagents, we concluded that Cys, Trp, Arg and Tyr seem to play important roles in the carbarylase activity of HSA. In addition, Tyr and Arg seem to be located in the active centre of the enzyme since carbaryl protected the activity from the inhibition. It was concluded that HSA hydrolyses carbaryl by a mechanism similar to that described for rabbit serum albumin based in transient carbamylation of a Tyr residue. The extrapolation of the hydrolysis rate to physiological albumin concentrations suggests that albumin might be playing a critical role in the detoxication of carbaryl.Abbreviations carbarylase carbaryl hydrolysing activity; - DCC N,N-Dicyclohexylcarbodiimide; - DEPC Diethyl pyrocarbonate; - DFP diisopropyl fluorophosphates; - DTNB 5,5-dithio-bis (2-nitrobenzoic acid); - HDCP O-hexyl O-2,5-dichlorophenyl phosphoramidate; - HS human serum;m - HSA human serum albumin; - NAI N-acetylimidazol; - NBS N-bromosuccinimide; - PNG phenylglyoxal; - p-NPB p-nitrophenyl butyrate     

Release of Human Serum Albumin from Poly(lactide-co-glycolide) Microspheres

Human serum albumin (HSA) was encapsulated in a 50:50 copolymer of DL-lactide/glycolide in the form of microspheres. These microspheres were used as a model formulation to study the feasibility of controlling the release of large proteins over a 20- to 30-day period. We show that HSA can be successfully incorporated into microspheres and released intact from these microspheres into various buffer systems at 37°C. A continuous release of the protein could be achieved in physiological buffers at 37°C over a 20- to 30-day period from microspheres with high protein loadings (11.6%). These results demonstrate the potential of poly(DL-lactide-co-glycolide) microspheres for continuous delivery of large proteins.     

重组人生长激素与人血清白蛋白治疗肝硬化低蛋白血症的成本-效果分析

目的：比较应用重组人生长激素和人血清白蛋白治疗肝硬化低蛋白血症的疗效、不良反应及成本一效果比。方法：运用药物经济学的成本一效果分析方法,对重组人生长激素和人血清白蛋白治疗肝硬化低蛋白血症的疗效及成本进行评价。结果：重组人生长激素和人血清白蛋白治疗肝硬化低蛋白血症,治疗后第４周有效率分别为６９．７０％、８４．８５％,成本一效果比分别为２７．３６、３８．８９;治疗后第８周有效率分别为７８．７９％、５４．５７％,成本一效果比分别为２４．２０、６０．４７。结论：重组人生长激素能有效治疗肝硬化低蛋白血症,能明显提高血清白蛋白含量并有较长的疗效;从远期看,成本－效果比优于人血清白蛋白。     

人血白蛋白临床应用调查及合理性分析

目的：调查某院人血白蛋白使用情况,为建立人血白蛋白合理使用管控体系提供依据和参考。方法：回顾性调查2013年1~6月份某院使用人血白蛋白的5257份病例并统计分析,随机抽取每月30份共180份病例进行合理性评价。结果：该院白蛋白使用率为11.8%,白蛋白费用占药品总费用9.4%。180份点评的病例中合理率46.1%。结论：该院人血白蛋白使用存在一定的不合理性,建议制定人血白蛋白合理使用规范,促进临床合理用药。     

阿司匹林与人血清白蛋白的相互作用研究

目的以光谱技术研究阿司匹林分子与人血清白蛋白(HSA)间结合作用机制。方法通过荧光光谱法确定阿司匹林对HSA的荧光猝灭机制。由Lineweaver-Burk双倒数作图法确定反应的解离常数。根据热力学方程讨论两者间主要的作用力类型。结合同步荧光技术考察阿司匹林对人血清白蛋白构象的影响。结果阿司匹林对HSA的荧光猝灭机制为静态猝灭。在37℃和25℃时阿司匹林与HSA的解离常数分别为KD37=1.44×10-3mol.L-1,KD25=1.96×10-3mol.L-1。结合反应热力学参数为ΔH=-19.73kJ.mol-1,△G=-16.21kJ.mol-1,ΔS=-11.77kJ.mol-1。结论两者结合的主要作用力类型是范德华力。阿司匹林与白蛋白结合后使蛋白质构象发生变化。     

荧光光谱法研究Ca2+存在下左氧氟沙星与人血清白蛋白的结合作用

目的：研究左氧氟沙星对人血清白蛋白以及在ca^2＋存在下左氧氟沙星与人血清白蛋白的结合作用。方法：通过荧光光谱法分析了左氧氟沙星对人血清白蛋白以及ca^2＋存在条件下左氧氟沙星对人血清白蛋白荧光淬灭光谱、同步荧光光谱,根据热力学方程讨论两者间主要的作用力类型。结果：在生理条件（pH=7．4,37℃）下,根据Stem—Volmer方程和荧光淬灭双倒数图,确定了左氧氟沙星对人血清白蛋白淬灭类型为静态淬灭,左氧氟沙星对人血清白蛋白的结合常数K=1．46×10^5L·mol^-1,结合位点n=1．1,根据热力学方法确定作用力类型为疏水作用力;在ca^2＋存在条件下,淬灭类型和作用力类型不变,结合常数K=2．38×10^4L·mol^-1,结合位点n：1．02。结论：在ca^2＋存在条件下,左氧氟沙星对人血清白蛋白的荧光淬灭减弱,结合常数和结合位点均变小。为研究左氧氟沙星的生物学效应,以及左氧氟沙星和Ca^2＋对蛋白质构象的影响等提供了重要信息。     

Effect of human serum albumin on transplacental transfer of glyburide

Glyburide is a second-generation sulfonylurea hypoglycemic drug used for the treatment of select women with pregestational and gestational diabetes mellitus (GDM). In vitro and in vivo investigations demonstrated its very low transplacental transfer to the fetal circulation. However, the factors influencing its low transfer across the human placenta remain unclear. Therefore, the goal of the current investigation was to determine the effect of human serum albumin (HSA) on the transfer and distribution of glyburide across the human placenta. To achieve this goal, the technique of dual perfusion of the placental lobule was utilized. The effect of HSA on the transfer of glyburide was determined at the range of glyburide to HSA molar ratios of 1:2-1:100. The transfer rate of free/unbound glyburide to the fetal circuit was 73+/-10% of the freely diffusible marker compound antipyrine (AP). Data obtained indicates the dependence of glyburide transfer and its retention by the placental tissue on the concentration of HSA.     

异硫氰酸荧光素标记人血清白蛋白

目的：用异硫氰酸荧光素（FTTC）对人血清白（HSA）进行荧光标记。方法：采用直接标记法,并考查不同料比时的荧光素分子与蛋白分子的结合情况,荧光标记物（FTTC－HSA）的荧光光谱及荧光寿命情况,结果：FTTC－HSA的激发波长为500nm,荧光波长为530nm;其固态和液态低温保存时荧光寿命分别为一年和37天。     

Interaction Between Two Dicarboxylate Endogenous Substances,Bilirubin and an Uremic Toxin, 3-Carboxy-4-Methyl-5-Propyl-2-Furanpropanoic Acid,on Human Serum Albumin

Purpose. Two dicarboxylate endogenous substances, bilirubin (BR) and 3-carboxy-4-methyl-5-propyl-2-furanpropanoic acid (CMPF), have a very high affinity to human serum albumin (HSA). This study was undertaken to clarify the existence of a dicarboxylate binding site on HSA. Methods. Chemical modification, pH dependent binding and X-ray crystallographic analysis were performed to characterize these dicarboxylate binding sites. Results. It was found the binding behavior for dicarboxylates was different from typical site I ligands such as warfarin (WF) and phenyl-butazone (PB) and that electrostatic interaction was an important factor for their binding to HSA. Moreover, His residues were considered to play an important role in pH dependent binding of dicarboxylic acids but in a different manner from the site I ligands. X-ray crystallography of CMPF and BR revealed the distances between the two carboxyl groups in their chemical structures were 5.854 Å and 9.979 Å, respectively. This difference may be reflected in pH dependent binding. Using fluorescent probe displacement, we attempted to identify the binding site for monocarboxylate derivatives of CMPF and investigated the role of individual carboxyl group in the recognition of the binding site. The results suggested two carboxyl groups were important for the specific binding of CMPF to site I. Conclusions. The binding site for dicarboxylic acids is located in subdomain IIA, which includes site I, on the HSA molecule. Electrostatic interaction is an important driving force for binding to HSA.