人和牛血清白蛋白的极谱测定

人或牛血清白蛋白溶液调至ＰＨ１０并在４℃左右放置３０小时,诱导出重复性良好的极谱峰,然后把试样溶液调至适当ＰＨ妆记录极谱电流,在浓度低于１．０×１０＾－５ｍｏｌ／Ｌ时,峰电流对浓度作图是直线,试样浓度可从图上直接读出,对影响白蛋白测定的诸因素作了详细研究和讨论,结果对富含硫的蛋白质极谱研究有参考价值。     

酪氨酸-铜体系电化学氧化行为的研究

在ｐＨ值３．９４的Ｂ－Ｒ缓冲溶液中,酪氨酸－铜络合物在玻碳电极出现氧化峰。用新极谱法测定,酪氨酸浓度在０．３μｇ／ｍｌ－３μｇ／ｍｌ范围内与氧化峰的２．５次微分值呈线性,检测下限０．２μｇ／ｍｌ。直接用于氨基酸药物样品的测定结果满意,并对酪氨酸－铜络合物的电化学氧化行为进行了研究。     

尿样中三种蛋白质的毛细管电泳分离检测方法研究

目的:建立毛细管电泳分离测定人尿样中转铁蛋白、白蛋白和血红蛋白的新方法.方法:通过选择运行缓冲溶液种类及浓度、pH、表面活性荆种类及浓度、分离电压、进样时间对蛋白质分离效果的影响,优化了毛细管电泳法分离转铁蛋白、白蛋白和血红蛋白的条件.结果:利用此方法测定三种蛋白质的含量,浓度在0.01到1.00 g L-1范围内与峰电流呈良好的线性关系,检出限均为10-4 g L-1.结论:所建立的方法用于人尿样中转铁蛋白、白蛋白和血红蛋白的测定,结果满意.     

三种因素对紫膜（PM）薄膜中菌紫质（BR）光反应类型的影响

由中性和碱性ｐＨ成膜液形成的ＰＭ薄膜在不同ＲＨ时ＣＤ语及其不同波长预光照产生的ＤＣＤ谱的研究发现,不同波长预光照产生的ＤＣＤ谱,按照在７００－４６０ｎｍ间的特征可分为全正型、长正短负型和短正长负型三大类。无论成膜液的ｐＨ是中性还是碱性,也不管ＰＭ薄膜的ＲＨ如何,≤４２８ｎｍ的近紫外光照都产生全正型ＤＣＤ谱,而≥６０ｎｍ的光照都产生短正长负型ＤＣＤ谱,但是５０６ｎｍ－５９０ｎｍ间的光照产生的ＤＣＤ谱却不仅与成膜液的ｐＨ有关,而且也与ＰＭ薄膜的ＲＨ有关,成膜液ｐＨ为１０．０的ＰＭ薄膜在ＲＨ７５％时未见有长正短负型ＤＣＤ谱。在近紫外的Ｍ峰区第一、三类ＤＣＤ谱无正峰,而在第二类中为正峰。这些特征都是ＰＭ薄膜中不同结构状态的ＢＲ分子配分浓度分布变化的反映。结果认为,成膜液的ｐＨ、膜所处的ＲＨ和预光照的波长均对ＢＲ的光循环反应有调控作用。这些信息可用于ＢＲ的调控研究,对ＢＲ分子器件的研制开发有重要的参考价值。     

示波极谱滴定法测定植物中钾含量的研究

示波极谱滴定法测定植物中的钾未见报道,本文用示波极谱滴定法对植物中钾的测定,做了多方面的研究。实验表明以ＨＡｃ－ＮａＡｃ为底液ｐＨ＝５．０的缓冲溶液中,加入过量的标准四苯研究钠（Ｎａ－ＴＰＢ）溶液,然后用标准ＡｆＮＯ３溶液滴定过量的四苯硼钠,利用四苯硼钠示波极谱曲线（ｄＥ／ｄｔ＝ｆ（Ｅ））上有切口,并和Ａｇ＋能定量反应,使切口消失为终点。测定结果满意。该法直观、简便、快速、灵敏、准确、不用加指示剂。溶液混浊、沉淀及有色,均不干扰测定,适用于植物中钾的测定。     

岩豆凝集素的圆二色性与生物学活性关系的研究

岩豆凝集素（ＭＤＬ）的远紫外圆二色性谱（ＣＤ谱）显示２１６－２１７ｎｍ处的单一负峰。此时ＭＤＬ分子含有１６．２％的α螺旋,４６．３％的β折叠和３７．５％的无规卷曲。ｐＨ９．０时负峰红移至２２０ｎｍ,且在２１７－２２２ｎｍ处的峰值几乎相同;在２０－４０℃范围内,ＣＤ谱的变化甚微;６０℃时谱峰蓝移;在８０℃或１００℃时,２１２ｎｍ处出现一大负峰。１ｍｏｌ／Ｌ或２ｍｏｌ／Ｌ脲时,ＭＤＬ的ＣＤ谱已发生明显变化,二级结构单元也有变化,凝集兔红细胞的活性也随之减弱;随脲浓度的增加,ＭＤＬ的谱峰蓝移,最终在２１２ｎｍ处出现大负峰。当胍浓度为０．７５ｍｏｌ／Ｌ时,ＭＤＬ的ＣＤ谱即有明显变化和活性丧失;胍浓度继续增加,ＣＤ谱逐渐成为特征的无规卷曲的谱形。在ｐＨ９．０、温度超过８０℃、脲或胍浓度分别高于２ｍｏｌ／Ｌ和０．７５ｍｏｌ／Ｌ时,ＭＤＬ的ＣＤ谱发生显著变化的同时,其凝集兔红细胞的生物学活性全部丧失,分子的二级结构单元也发生很大改变。     

金属离子对盐酸多西环素与牛血清白蛋白的结合常数的影响

利用荧光光谱法、紫外光谱法研究了盐酸多西环素与牛血清白蛋白的作用。还研究了不同浓度的Ca2+、Mg2+、Zn2+、Cu2+、Fe3+存在时,对盐酸多西环素与血清白蛋白的相互作用的影响。金属离子的存在能够影响盐酸多西环素与BSA结合,使其反应的表观结合常数值降低了,并且反应常数随Ca2+、Mg2+、Zn2+浓度的增加而降低。     

脲对果菠萝蛋白酶活力与构象的影响

本文用荧光光谱,紫外差示光谱和CD谱研究果菠萝蛋白酶在不同浓度的脲溶液中的构象及酶活力的变化情况。酶的荧光强度随脲浓度增大而明显增加,8mol/L脲使荧光强度增强65％,发射峰出现红移。差示谱表明在232nm和288nm出现二个正峰,它们均随脲浓度增大而加剧,前者与主链构象变化有关,而后者与生色基团(Trp、Tyr)的微环境变化相关。CD谱表明:天然酶在208nm和225nm处有二个负峰,脲变性后,225nm的负峰基本上不随脲浓度增大而变化,但208nm峰则明显发生变化并逐渐出现红移,6mol/L以上此峰则完全消失。     

白蛋白对低粘切变流中的红细胞流变特性的影响

用一种新型激先衍射法,研究了白蛋白对低粘切交流动中红细胞取向的影响。所得结果表明,当白蛋白浓度小于０．０４ｇ／Ｌ时,测得的取向指数（ＤＩ）ｏｒ随蛋白浓度迅速增加,在白蛋白浓度等于０．０４ｇ／Ｌ时,其取向指数（ＤＩ）ｏｒ达到最大值,以后又随白蛋白浓度的增加而很快降低到一稳定值（白蛋白浓度大于０．５ｇ／Ｌ时）。同时作者从流变学观点对上述实验结果作了解释,并通过实验证明了取向指数可用于描述红细胞变形性,从而证明了在一定浓度下（０．０４ｇ／Ｌ）白蛋白有增加红细胞变形性,保持红细胞双凹圆盘形的功能。该研究提示我们,在用此新型激光衍射法时,必须在介质中加入适量的白蛋白（０．０４ｇ／Ｌ）。     

荧光法研究血清白蛋白与药物的结合作用

本文应用荧光光谱法,观测了药物分子头孢菌素Ⅳ、异烟肼、维生素B_6和氟哌酸对白蛋白荧光的猝灭。由Lineweaver-Burk双倒数作图法,确定了药物与白蛋白作用的离解常数。并通过Forster偶极-偶极无辐射能量转移机理确定了药物分子氟哌酸在人血清白蛋白中与色氨酸残基之间的距离R为2.55nm,由这一距离确定了药物分子可能进入的区域和位置。     

Electrochemistry of 2-thio- and 2-oxo-pyrimidines in dimethyl formamide in the presence of dioxygen

Electrocatalysis of dioxygen reduction to superoxide ion in dimethylformamide is afforded by a reversible interaction with thiol or hydroxo forms of pyrimidine which takes place only on the mercury surface electrode even in the absence of transition metal ions. The interaction is revealed by a polarographic wave and by a voltammetric peak which can be recorded only in the presence of dioxygen in the pyrimidines solution. The polarographic wave and voltammetric peak are attributed to the reduction of the oxygenated pyrimidine. In the range of concentration where the polarographic limiting currents due to the reduction of the oxygenated and unoxygenated pyrimidine are of the same order of magnitude, the stoichiometric ratio dioxygen/pyrimidine is determined to be 1. The electrode process probably involves the formation of a complex of mercury with the thiol or the hydroxo form of the pyrimidine on the electrode surface. Adsorption or coadsorption of the reactants seems to be involved.     

Single-sweep polarographic investigation of concanavalin A and its interaction with polysaccharide.

A simple, rapid, and sensitive single-sweep polarographic method has been developed for investigation of concanavalin A (con A) and its interaction with selected polysaccharides. In a solution containing 0.001 M 2,2'-bipyridine, 0.015 M hexamethylenetetramine, and 0.1 M sodium chloride, con A exhibits a single-sweep polarographic wave, and the cathodic peak potential is -1.50 V (vs SCE). The peak current varies linearly with con A concentration over a range of 1.0 x 10(-8) to 1.2 x 10(-7) M by derivative single sweep polarography. A preliminary discussion on properties of the con A polarographic wave has been made. In addition, it has been demonstrated that single sweep polarography can be a useful method for studies on interactions of con A with its complementary polysaccharides in solution.     

Immobilized antibody-based flow type enzyme immunosensor for determination of human serum albumin

A flow-type enzyme immunosensor was prepared for the electrochemical determination of human serum albumin (HSA). The immunosensor was constructed from the immobilized antibody (anti-HSA IgG) reactor and an oxygen electrode. The immunochemical reaction of catalase-labelled antibody with HSA was completed with 30 min. After the immunochemical reaction, hydrogen peroxide solution was injected into the system and a peak current was obtained within 2 min. A linear relationship was observed between the current increase and the logarithm of HSA concentration in the range 10⁻⁸-10⁻⁶ g ml⁻¹. The minimum measurable concentration was 10⁻⁸ g ml⁻¹. The current increase was reproducible with 10% of the relative errors when a sample solution containing 10⁻⁷ g ml⁻¹ of HSA was used. The minimum measurable concentration increased to 10⁻⁹ g ml⁻¹ when hydrogen peroxide was recycled for 5 min in the reactor system. The immobilized antibody reactor could be reused. HSA in human serum was determined by the system proposed.     

A simple differential pulse polarographic method for the determination of thymoquinone in black seed oil

A reliable and simple differential pulse polarographic method is described for the determination of thymoquinone in black seed oil. The polarographic behaviour of thymoquinone was examined in various buffer systems over the pH range 5.0-10.0. Thymoquinone is reduced in a single, reversible peak at the dropping mercury electrode. The differential pulse polarogram showed a distinct peak in S?rensen buffer:methanol (3:7, v/v; pH 8.5) at a peak potential of -0.095 V (vs. silver/silver chloride electrode), and a plot of peak height against concentration was found to be linear over the range 0.2-15.0 microg/mL (R = 0.9998). The limit of detection was calculated to be 0.054 microg/mL. The polarographic method has been applied to determine thymoquinone in two black seed oil preparations available on the Austrian pharmaceutical market.     

Polarographic direct determination and homogeneous immunoassay of anti-human serum albumin (HSA) by parallel catalytic hydrogen wave of anti-HSA or HSA

Human serum albumin (HSA) or anti-human serum albumin (anti-HSA) yields a catalytic hydrogen wave at about -1.85V (vs Ag/AgCl) in 0.25M NH(3).H(2)O-NH(4)Cl (pH 8.58) buffer. When 1.0 x 10(-2)M K(2)S(2)O(8) is present, the catalytic hydrogen wave is further catalyzed, producing a parallel catalytic wave of hydrogen as catalyst in nature, termed the parallel catalytic hydrogen wave. The sensitivity of the parallel catalytic hydrogen wave is higher by two orders of magnitude than that of the catalytic hydrogen wave. Using the parallel catalytic hydrogen wave of anti-HSA or HSA in the presence of K(2)S(2)O(8), two sensitive methods for the determination of anti-HSA were developed. One is a direct determination based on the parallel catalytic hydrogen wave of anti-HAS itself, and the other is a homogeneous immunoassay based on measuring the decrease of the peak current of the parallel catalytic hydrogen wave of HSA after homogeneous immunoreaction of HSA with anti-HSA. In the direct determination, the second-order derivative peak current of the parallel catalytic hydrogen wave of anti-HSA itself is rectilinear to its titer in the range from 1:1.0 x 10(7) to 1:8.4 x 10(6). In the homogeneous immunoassay, the decrease in the second-order derivative peak current of the parallel catalytic hydrogen wave of HSA is linearly related to the added anti-HSA in the titer range from 1:3.0 x 10(7) to 1:6.0 x 10(6). These assays are highly sensitive and rapid in operation and can be used to evaluate such antigens and their antibodies as those that could yield the parallel catalytic hydrogen wave.     

从反胶束溶液中反萃牛血清白蛋白的研究

由近红外光谱证实,十六烷基三甲基溴化铵(CTAB)/正己醇-正辛烷反胶束溶液是牛血清白蛋白(BSA)增溶于非极性有机溶剂的较理想中介。研究了反萃液酸度、离子强度和种类等参数对BSA反萃率的影响,过低的溶液酸度导致BSA变性。适宜的反萃液为:1.0～2.0mol/L KBr,pH4.3～4.9。蛋白质的紫外光谱表明,BSA分子在料液,反胶束溶液和反萃液中的构象大致相同。此外,探讨了反胶束溶液循环使用的效率问题。最后,通过采用适当的相比,成功地实现了蛋白质的回收和浓缩。     

Adsorption of BSA on QAE-dextran: equilibria

Equilibrium isotherms for adsorption of bovine serum albumin (BSA) on a strong-base (QAE) dextran-type ion exchanger have been determined experimentally. They were not affected by the initial concentration of BSA but were affected by pH considerably. They were correlated by the Langmuir equation when pH >/= 5.05 and by the Freundlich equation of pH 4.8, which is close to pl approximately 4.8 of BSA. The contribution of ion exchange to adsorption of BSA on the ion exchanger was determined experimentally. The maximum amounts of inorganic anion exchanged for BSA were 1% and 0.4% of the exchange capacity of the ion exchanger at pH 6.9, respectively. Since the effect of the ion exchange on the adsorption appeared small, BSA may be adsorbed mainly by electrostatic attraction when pH >/= 5.05 and by hydrophobic interaction or hydrogen bonding at pH 4.8. When NaCl coexisted in the solution, the shape of the isotherm was similar to the Langmuir isotherm, but it is shifted to the right. When the concentration of NaCl was 0.2 mol/dm(3), BsA was not adsorbed on the resin. When BSA was dissolved in pure water, the saturation capacity of BSA on HPO(4) (2-),-orm resin was about 2 times larger than that for adsorption from the solution with buffer (pH 6.9 and 8.79). The saturation capacity for adsorption of BSA in pure water on HPO(4) (2-) + H(2)O(4) (-)-from resin was much smaller than that from the solution with buffer. The isotherms for univalent Cl(-)-and H(2)PO(4) (-)-form resin was peculiar; that is, the amount of BSA adsorbed decreased with increasing the liquid-phase equilibrium concentration of BSA. (c) 1993 John Wiley & Sons, Inc.     

Mesostructure of fibrillar bovine serum albumin gels

The mesostructure of bovine serum albumin (BSA) at low pH was investigated. Rheological measurements were performed to determine the critical percolation concentration (c_p). A decreasing c_p with increasing ionic strength was found. Fibrils with a contour length of about 100–300 nm were found using transmission electron microscopy. The measured conversion of monomers into fibrils was independent of ionic strength (0.20–0.30 M). Dilution of BSA samples showed that the aggregation process is reversible and that there exists a critical concentration for the self-assembly of BSA. We explain the decreasing c_p with increasing ionic strength in terms of an adjusted random contact model.     

A novel in vitro system for gamete fusion in maize

Various systems by using electric pulse, calcium, or polyethylene glycol have been developed in the past decade for the in vitro fusion of plant gametes. These in vitro systems provide a new way to study the fertilization mechanisms of plants. In this study, we developed a bovine serum albumin （BSA）-mediated fusion system for the in vitro fusion of maize gametes. The in vitro fusion of the isolated single egg cell and sperm cell of maize was observed microscopically in the BSA solution and the fertilized egg cell showed normal cell wall regeneration and nuclear division. The effects of the BSA concentration, pH value and calcium level on the efficiency of the maize gamete fusion were also assessed. BSA concentration and pH value did significantly affect the efficiency of the gamete fusion. Calcium was not necessary for the gamete fusion when BSA was present. The optimal solution for the gamete fusion contained 0.1% BSA, pH 6.0. The fusion frequency was as high as 96.7% in that optimal solution. This new in vitro fertilization system offers an alternative tool for the in vitro study of fertilization mechanisms with much simpler manipulating procedure than PEG system, and it will be especially useful for the in vitro study of the calcium dynamics during plant fertilization.     

The subdomain structure of human serum albumin in solution under different pH conditions studied by small angle X-ray scattering

Small-angle X-ray scattering (SAXS) was used to study structural characteristics of human serum albumin (HSA) in solution under different pH conditions. Guinier analysis of SAXS results yielded values of the molecular radius of gyration ranging from 26.7 Å to 34.5 Å for pH varying from 2.5 to 7.0. This suggests the existence of significant differences in the overall shape of the molecule at different pH. Molecular models based on subdomains with different spatial configurations were proposed. The distance distribution functions associated with these models were calculated and compared with those determined from the experimental SAXS intensity functions. The conclusion of this SAXS study is that the arrangement of molecular subdomains is clearly pH dependent; the molecule adopting more or less compact configuration for different pH conditions. The conclusions of this systematic study on the modification in molecular shape of HSA as a response to pH changes is consistent with those of previous investigations performed for particular pH conditions. Correspondence to: J. R. Olivieri