应用明矾——活性炭吸附法去除静脉注射用人血免疫球蛋白制 …

用活性炭、硅藻土、氢氧化铝、明矾－活性炭四种不同的吸附剂处理静脉注射用人血免疫球蛋白制剂。结果表明,明矾－活动性吸附法去除热原质最有效,同时进一步改善了制品外观,提高了制品纯度,蛋白回收率达到７５％以上,而且毒性试验和安全试验也符合标准。铝离子残余量偏低。     

抗狂犬病血清热原处理工艺的试验研究

作者对抗狂犬病血清精制工艺中明矾吸附步骤进行了试验研究。结果表明,采用合理的吸附剂浓度、适当的吸附时间,对制品中热原质的去除效果明显。改进工艺应用于大批量生产,取得满意效果。     

菜籽降压肽双酶水解液脱色工艺的研究

用活性炭对菜籽降压肽双酶水解液进行脱色处理,采用单因素试验以及单因素试验基础上的正交试验法。考察活性炭用量、pH、温度、时间对水解液脱色率、肽损失率、活性及ACE抑制率的影响。正交试验优化结果表明,菜籽降压肽双酶水解液脱色最佳工艺为活性炭的用量为1%、脱色pH值为3.0、脱色温度为80℃、脱色时间为50 min,在此条件下脱色率为85.52%、ACE抑制率为57.83%、活性损失率为5.69%。     

中国小学生乳类及制品消费情况分析

目的：分析2016—2017年我国小学生乳类及制品的消费状况。方法：分析数据来自“2016—2017年中国儿童与乳母营养健康监测”，抽样方法采用多阶段分层随机抽样，研究对象为全国275个监测点4 2930名小学生。采用食物频率法的膳食调查问卷，收集小学生的乳类及制品消费情况。结果：2016—2017年我国小学生乳类及制品的消费率为80%，男生为79.5%、女生为80.5%，城市为89%、农村为71.9%，大城市、中小城市、普通农村和贫困农村、乳类及制品的消费率分别为93.2%、87.9%、73%、69.2%。液态奶（53.7%）和酸奶（59.5%）的消费率相对较高。2016—2017年我国小学生消费乳类及制品的频次≥1次/d、4~6次/w、1~3次/w、<1次/w、不消费的百分比分别为41.7%、13.3%、21.9%、3.2%、19.8%，≥1次/d百分比最高。消费乳类及制品的小学生，液态奶、奶粉、酸奶和其他乳制品的日均消费量分别为154.7、46.6、92.4、23.7 g。将所有乳类及制品的消费量折合成液态奶的消费量，2016—2017年我国消费乳类及制品的小学生折合液态奶的消费量为198 g/d，城市（230.9 g/d）高于农村（161.2 g/d）；大城市、中小城市、普通农村、贫困农村分别为264.4、220.2、164.1、153.4 g/d，呈现下降趋势。2016—2017年我国小学生乳类及制品消费量达到《中国居民膳食指南》（2016）推荐摄入量的比例为13.3%，在消费人群中为16.6%。结论：2016—2017年我国小学生乳类及制品消费水平整体偏低，尤其是农村地区，需进一步加强对小学生及其家长相关的健康教育，增加小学生乳类及制品的消费水平。     

熟地多糖活性炭脱色工艺的研究

本研究旨在探讨活性炭对熟地多糖提取液脱色的工艺条件。在单因素试验基础上,采用正交试验,以多糖脱色率(%)和多糖剩余率(%)为指标,用紫外-可见分光光度法测定,确定了熟地多糖活性炭脱色的最优工艺的参数。结果发现:活性炭添加量对脱色效果的影响最大,其次为温度,再次为时间。最佳脱色条件为脱色温度60℃,活性碳添加量5%,吸附时间30 min,在此条件下,脱色率为98.20%,多糖剩余率为84.89%。从而表明活性炭对熟地多糖脱色工艺可行,简便,有效。     

免疫球蛋白的裂解

淀粉凝胶电泳是检查裂解的最好工具。裂解和聚集作用一样都可以用交联葡聚糖凝胶G—200层析测定。这些实验可以检查制品在效期内不同时间的聚集和裂解。产品最终的PH值最好保持在允许的下限6.4,这样r—球蛋白不易发生裂解。加入抑制蛋白分解的抑制剂,例如EACA和TrasyLoL需进一步试验,最后关于确定产品的聚集和裂解的最大允许程度,尚需进一步试验。     

丙种球蛋白的抗体测定和裂解情况

众所熟知,利用人胎盘血或血浆提制的丙种球蛋白是含有特异性抗体能够防治多种传染病的蛋白制剂。这种制剂所含的抗体种类和抗体水平,取决于不同地区或同一地区不同时期提供胎盘或血液来源的人群免疫状况;而抗体的稳定性则与制品在保存中的裂解情况有密切关系。我所出品的丙种球蛋白,系收集兰州、西安、银川、宝鸡等地区的胎盘,按硫酸铵盐析——明矾吸附法进行生产的。为了考查本所丙球制品有关效力方面     

评价肺炎支原体疫苗效力的仓鼠攻击试验

<正> 在1964-1976年间,许多实验室研制了一系列的福尔马林灭活的肺炎支原体疫苗并进行了效力试验。有些疫苗是在众多的部队新兵中进行了现场试验,有些则给少数志愿受试者接种,然后用强毒株进行攻击试验。利用接种过疫苗者中支原体肺炎自然发病率的降低和感染过支原体人群中病情的减轻来进行评价,结果表明,这些疫苗的保护效力是很低的。当将疫苗加入明矾佐剂增强其抗原性或简单地增加疫苗剂量时,能提高用     

固氮菌菌剂的应用

我们从1972年开始进行花生、大豆根瘤菌剂的应用试验,取得了较好的效果,现分别介绍如下: (一) 花生根瘤菌剂的试验 花生根瘤菌菌剂是用湖北省油料研究所提供的007、009两个菌株,试制了草炭制品13袋,应用15亩。因受旱欠收,但从验收的春胜大队九小队一块试验田的实收产量看,拌菌较对照增长10.39％,通过重复三点取样,每点3—5株考查了经济性状,拌菌花生较     

一种纯化细菌有机磷水解酶的简便方法

Cibacron blue T_3GA与溴化氰活化的Sepharose 4B偶联后,产生一种能有效地分离有机磷水解酶的吸附剂。用0.15mol/L MgCl_2溶液从黄杆菌P3—2细胞抽提出的粗酶液通过柱层析分离,即可得到纯化8倍、酶活性回收率为269.4％的纯酶制品。该酶制品用凝胶电泳测是均一的。     

采用多种抗原测定静注人免疫球蛋白(pH4)中抗体Fc段生物学活性的初探

目的初步探讨采用多种抗原测定静注人免疫球蛋白(IVIG)(pH4)中抗体的Fc段生物学活性,了解IVIG中抗体的Fc段生物学活性。方法采用补体活化的经典途径中免疫复合物激活补体的方法,将不同浓度的麻疹病毒、风疹病毒、乙肝表面抗原(HBsAg)、破伤风类毒素、脑膜炎球菌P64k外膜蛋白和白喉类毒素6种抗原分别致敏人O型血红细胞形成红细胞-抗原结合物;然后,6种致敏红细胞分别与IVIG孵育,与特异性抗体形成红细胞-抗原-抗体复合物;最后,此复合物与补体反应,在541 nm波长处读取吸光值,并绘制溶血反应动力学曲线,分别计算IVIG中针对上述6种抗原IgG的Fc段生物学活性。采用此方法,用6种抗原致敏的红细胞测定IVIG的Fc段生物学活性10次,验证此方法的重复性。结果麻疹病毒、风疹病毒、HBsAg、破伤风类毒素和脑膜炎球菌P64k外膜蛋白致敏的红细胞分别与供试品和补体反应后,测定的溶血反应动力学曲线较平缓,而白喉类毒素致敏的红细胞与供试品和补体反应后,测定的溶血反应动力学曲线下降明显,呈典型的"S"型曲线。计算结果显示,IVIG中针对此六种抗原的抗体Fc段生物学活性相对于参考品均大于80%。Fc段生物学检测方法重复性较好。结论采用多种抗原分别致敏红细胞,可以用来检测IVIG中多种抗体的Fc段生物学活性,为深入了解IVIG制品中的多种抗体的Fc段生物学活性奠定了基础。     

Managing ammonia emissions from dairy cows by amending slurry with alum or zeolite or by diet modification

Animal agriculture is a significant source of atmospheric ammonia. Ammonia (NH3) volatilization represents a loss of plant available N to the farmer and a potential contributor to eutrophication in low-nitrogen input ecosystems. This research evaluated on-farm slurry treatments of alum or zeolite and compared three diets for lactating dairy cows in their effectiveness to reduce NH3 emissions. NH3 emissions were compared using a group of mobile wind tunnels. The addition of 2.5% alum or 6.25% zeolite to barn-stored dairy slurry reduced NH3 volatilization by 60% and 55%, respectively, compared to untreated slurry. The alum conserved NH3 by acidifying the slurry to below pH 5, while the zeolite conserved ammonia by lowering the solution-phase nitrogen through cation exchange. The use of alum or zeolite also reduced soluble phosphorus in the slurry. NH3 loss from fresh manure collected from lactating dairy cows was not affected by three diets containing the same level of crude protein but differing in forage source (orchardgrass silage vs. alfalfa silage) or neutral detergent fiber (NDF) content (30% vs. 35% NDF). NH3 losses from the freshly excreted manures occurred very rapidly and included the urea component plus some unidentified labile organic nitrogen sources. NH3 conservation strategies for fresh manures will have to be active within the first few hours after excretion in order to be most effective. The use of alum or zeolites as an on-farm amendment to dairy slurry offers the potential for significantly reducing NH3 emissions.     

Evidence for the denaturation of recombinant hepatitis B surface antigen on aluminium hydroxide gel

Despite the complexity of the subject of protein–alum interactions, a valuable information can be obtained by analyzing the adsorbed and desorbed protein by common physico–chemical methods. In the present work, to approach the adsorption of hepatitis B surface antigen (HBsAg) on alum, the experimental data were supported by complementary analyses of the adsorbed protein by immunoelectron microscopy and the desorbed protein by denaturing size-exclusion chromatography and sodium dodecyl sulfate-polyacrylamide gel electrophoresis under reducing conditions. First, the depletion of HBsAg was investigated. The aspects assessed were the conditions, recovery and chromatographic performance of the desorbed protein. The results obtained strongly suggested the loss of particulate structure of HBsAg after adsorption on alum. This conclusion was further reinforced by direct immunoelectron microscopic visualization of HBsAg in the adsorbed state.     

Priming of immune responses to hepatitis B surface antigen in young mice immunized in the presence of maternally derived antibodies

Early vaccination is necessary to protect infants from various infectious diseases. However, this is often unsuccessful largely due to the immaturity of the neonatal immune system. Furthermore, maternally derived antibodies can interfere with active immunization. We have previously shown in young mice that immune responses against several different antigens can be improved by the addition of oligodeoxynucleotides containing immunostimulatory CpG motifs (CpG ODN). In this study we have evaluated immunization of newborn (1-7-day-old) BALB/c mice against hepatitis B surface antigen (HBsAg), with alum and/or CpG ODN, in the presence of high levels of maternal antibody against HBsAg (anti-HBs). Seroconversion rates and anti-HBs titers were compared to those induced by a HBsAg-expressing plasmid, since other studies had suggested DNA vaccines to be superior to protein vaccines in young mice with maternal antibody. HBsAg/alum/CpG ODN was superior to DNA vaccine in inducing HBsAg-specific CTL responses in young mice in the presence of maternally transferred anti-HBs antibodies. However, B cell responses to both HBsAg/alum/CpG ODN and DNA vaccines remained weak in the presence of maternally transferred anti-HBs antibodies.     

Monitoring of the serum proteome in Kawasaki disease patients before and after immunoglobulin therapy

Kawasaki disease (KD) is a systemic vasculitis that mainly affects children younger than 5 years. The causal pathogen is unknown, therefore specific diagnostic biomarkers and therapy are unavailable. High-dose intravenous immunoglobulin (IVIG) is considered as the most effective therapy to reduce the prevalence of coronary artery lesion (CAL) in KD; however, it has side effects. This study aimed to (1) determine whether IVIG therapy is effective at the molecular level; (2) provide the first serum proteomic profile of KD under IVIG therapy; and (3) screen for monitoring biomarker candidates. We extracted serum proteins from samples of healthy individuals and from KD patients before and after IVIG therapy, and employed two-dimensional electrophoresis and MALDI-TOF/TOF mass spectrometry to identify differentially expressed proteins. The identifications were validated by Western blotting. We identified 29 differentially expressed proteins in KD patients and found that IVIG therapy restored most of these proteins to near-normal levels. Tracing the protein levels of single patients before and after IVIG therapy showed that the proteins, especially Transthyretin (TTR), are potential markers for therapeutic monitoring. Functional analyses of these proteins by PANTHER and String suggested that the key influence of KD lay in the immune system, which was targeted by IVIG.     

Biothermodynamic analysis of BSA adsorption to alum gel using isothermal titration calorimetry

In this study, we used ITC (isothermal titration calorimetry) to quantitatively investigate the impacts of temperature and protein concentration on adsorption behavior on a solid surface, using BSA (bovine serum albumin) as a model protein, and alum (aluminum hydroxide) gel as an adsorbent. The zeta potential measurement for alum gel (0.25 mV at pH 9.3) revealed that its surface charge was not strong enough for electrostatic interaction. ITC analysis showed that the BSA-alum gel interaction was entropy-driven, suggesting that during adsorption, water molecules were expelled from the hydration layers of the alum gel and BSA. Therefore, the major mechanism for the BSA-alum gel interaction was hydrophobic interaction rather than electrostatic interaction. This biothermodynamic approach can be helpful not only to identify interaction mechanisms, but also to explore the optimum conditions for protein-adsorbent interactions.     

Pretreatment with alum or powdered activated carbon reduces bacterial predation-associated irreversible fouling of membranes

This study evaluated the co-application of bacterial predation by Bdellovibrio bacteriovorus and either alum coagulation or powdered activated carbon adsorption to reduce fouling caused by Escherichia coli rich feed solutions in dead-end microfiltration tests. The flux increased when the samples were predated upon or treated with 100 ppm alum or PAC, but co-treatment with alum and predation gave the best flux results. The total membrane resistance caused by the predated sample was reduced six-fold when treated with 100 ppm PAC, from 11.8 to 1.98 × 10¹¹ m^?1, while irreversible fouling (R_p) was 2.7-fold lower. Treatment with 100 ppm alum reduced the total resistance 14.9-fold (11.8 to 0.79 × 10¹¹ m^?1) while the R_p decreased 4.25-fold. SEM imaging confirmed this, with less obvious fouling of the membrane after the combined process. This study illustrates that the combination of bacterial predation and the subsequent removal of debris using coagulation or adsorption mitigates membrane biofouling and improves membrane performance.     

Immunotherapy approach with recombinant survivin adjuvanted with alum and MIP suppresses tumor growth in murine model of breast cancer

Survivin has received attention as a potential target for cancer immunotherapy because of its crucial role in oncogenesis. We undertook this study to evaluate the immunotherapeutic potential of combination of recombinant survivin along with adjuvant alum and immune modulator Mycobacterium indicus pranii (MIP). In vivo efficacy of the combination was studied in an invasive murine breast cancer model. Recombinant survivin protein was purified from Escherichia coli based expression system and characterized by western blotting. Purified survivin protein was combined with alum and MIP and was used for immunization of Balb/c mice. Antigen-primed animals were then challenged with syngeneic mammary tumor cells known as 4T-1. Balb/c mice spontaneously develop tumor when inoculated with 4T-1 cells. Antigen and adjuvant combination was immunogenic and significantly suppressed tumor growth in mice immunized with combination of recombinant survivin (10?µg), alum, and MIP. This is the first report that describes a combination immunotherapy approach using recombinant survivin, alum, and MIP in highly metastatic murine breast cancer model and holds promise for development of new biotherapeutics for cancer.     

l-Proline reduces IgG dimer content and enhances the stability of intravenous immunoglobulin (IVIG) solutions

Liquid intravenous immunoglobulin (IVIG) products offer improved convenience in preparation but often lack sufficient stability to allow room temperature storage. Furthermore, clinical tolerability may be affected due to formation of idiotype/anti-idiotype IgG dimers and/or aggregates. Here we report on the development of a 10% IVIG formulation with optimized stability achieved by the use of l-proline. The stability of concentrated liquid IVIG was strongly pH dependent. Aggregate formation, yellowish discoloration of the solution and loss of anti-hepatitis B surface antigen (HBs) antibody activity was minimal at intermediate pH (pH 4.8–5.3). Fragmentation of IgG was highest at low pH (pH 4.1). Idiotype/anti-idiotype IgG dimer formation was highest at neutral pH and was reduced with decreasing pH. The presence of l-proline further improved stability by inhibiting protein aggregation, reducing loss of anti-HBs antibody activity and decreasing coloring, particularly compared with glycine formulations. The IgG dimer content was up to 30% lower in solutions containing l-proline compared with those containing glycine or other stabilizers. In conclusion, a weakly acidic pH of approximately 5 and l-proline as stabilizer are optimal conditions for long-term stability of a liquid IVIG. l-proline, an amphiphilic, naturally occurring amino acid, is superior to glycine in restricting IgG dimer formation.     

A Novel HIV Vaccine Adjuvanted by IC31 Induces Robust and Persistent Humoral and Cellular Immunity

The HIV vaccine strategy that, to date, generated immune protection consisted of a prime-boost regimen using a canarypox vector and an HIV envelope protein with alum, as shown in the RV144 trial. Since the efficacy was weak, and previous HIV vaccine trials designed to generate antibody responses failed, we hypothesized that generation of T cell responses would result in improved protection. Thus, we tested the immunogenicity of a similar envelope-based vaccine using a mouse model, with two modifications: a clade C CN54gp140 HIV envelope protein was adjuvanted by the TLR9 agonist IC31®, and the viral vector was the vaccinia strain NYVAC-CN54 expressing HIV envelope gp120. The use of IC31® facilitated immunoglobulin isotype switching, leading to the production of Env-specific IgG2a, as compared to protein with alum alone. Boosting with NYVAC-CN54 resulted in the generation of more robust Th1 T cell responses. Moreover, gp140 prime with IC31® and alum followed by NYVAC-CN54 boost resulted in the formation and persistence of central and effector memory populations in the spleen and an effector memory population in the gut. Our data suggest that this regimen is promising and could improve the protection rate by eliciting strong and long-lasting humoral and cellular immune responses.