血清4型禽腺病毒Fiber-1蛋白截短表达及多克隆抗体制备

目的:制备血清4型禽腺病毒(FAdV-4)Fiber-1基因截短蛋白多克隆抗体,为FAdV-4病的诊断、检测及致病机制研究奠定基础。方法:对FAdV-4的CH/AHMC/2015分离株Fiber-1基因序列(MG148335.1:30459-31754)进行信号肽、疏水性和抗原决定簇分析,截取具有较高免疫原性的片段,设计合成特异性引物,以CH/AHMC/2015分离株基因组DNA为模板进行PCR扩增,获得截短Fiber-1(sFiber-1)基因片段,将其连接至原核表达载体pET-32a,验证后转化至大肠杆菌BL21感受态中,诱导表达sFiber-1蛋白。纯化后的蛋白与佐剂乳化后免疫SD大鼠,制备多克隆抗体,并测定多克隆抗体效价。用Western-blot检测抗体免疫原性。结果:成功构建了sFiber-1的原核表达载体,获得FAdV-4的sFiber-1重组蛋白,经SDS-PAGE鉴定,重组sFiber-1蛋白大小约为52 kDa,主要以包涵体形式表达;间接ELISA法测得sFiber-1多克隆抗体效价为1∶2^(13);Western blot结果显示制备的多克隆抗体能特异性识别出重组sFiber-1蛋白。结论:本研究成功表达了FAdV-4的重组sFiber-1蛋白,制备了具有较高免疫活性的sFiber-1多克隆抗体,可为FAdV-4的检测及诊断奠定基础。

Giving Food a Fiber Boost: Adding High Beta-Glucan Ingredient from Barley to Everyday Foods

Lack of dietary fiber contributes to many health issues, particularly chronic vascular diseases. Mixed linkage β-1.3 - 1.4 beta-glucan (beta-glucan, in this paper) is a confirmed beneficial ingredient for the human diet through reduction of cholesterol and the glycemic index. Barley contains the highest beta-glucan content of all the grains, and in this study, a percentage of flour from two high beta glucan lines was, each, added to an array of wheat-based food products to measure how it impacted total dietary fiber. Results showed that beta-glucan content was higher in all the products containing the added high beta-glucan flour, along with increased total dietary fiber content. Protein content in the food products is also increased with the higher protein in the barley flours added. Beta-glucan content in the barley-added products increased to 1.2% - 4.0% versus 0.2% - 0.5% in the pure wheat products, while the dietary fibers increased to 3.5% - 24.4% versus 2.1% - 9.1% in pure wheat product controls. This research provided experimental evidence that using a high beta-glucan barley ingredient in food can increase dietary fiber to benefit health.

Research on the Development of Fibroin and Nano-Fiber from Silk Cocoons for Regenerated Tissue Engineering Applications by Electro-Spinning

In this paper, the main goal is to prepare silk fibroin nano-fiber, which is used for regenerated tissue applications. Silk scaffold nano-fibers made by electro-spinning technology can be used in regenerated tissue applications. The purpose of the research is to prepare a silk-fibroin nano-fiber solution for potential applications in tissue engineering. Using a degumming process, pure silk fibroin protein is extracted from silk cocoons. The protein solution for fibroin is purified, and the protein content is determined. The precise chemical composition, exact temperature, time, voltage, distance, ratio, and humidity all have a huge impact on degumming, solubility, and electro-spinning nano-fibers. The SEM investigates the morphology of silk fibroin nano-fibres at different magnifications. It also reveals the surface condition, fiber orientation, and fiber thickness of the silk fibroin nano-fiber. The results show that regenerated silk fibroin and nano-fiber can be used in silk fibroin scaffolds for various tissue engineering applications.

Optimization of the Pretreatment of the Mixture of Cassava Peelings and Pineapple Fibers Using Response Surface Methodology and a Process Simulator for the Bioethanol Production

The increase in oil prices and greenhouse gas emissions has led to the search for substitutes for fossil fuels. In Cameroon, the abundance of lignocellulosic resources is inherent to agricultural activity. Production of bioethanol remains a challenge given the crystallinity of cellulose and the presence of the complex. The pretreatment aimed to solubilize the lignin fraction and to make cellulose more accessible to the hydrolytic enzymes, was done using the organosolv process. A mathematical modeling was performed to point out the effect of the temperature on the kinetics of the release of the reducing sugars during the pretreatment. Two mathematical model was used, SAEMAN’s model and Response surface methodology. The first show that the kinetic parameters of the hydrolysis of the cellulose and reducing sugar are: 0.05089 min-1, 5358.1461 J·mol-1, 1383.03691 min-1, 51577.6100 J·mol-1 respectively. The second model was used. Temperature is the factor having the most positive influence whereas, ethanol concentration is not an essential factor. To release the maximum, an organosolv pre-treatment of this sub-strate should be carried out at 209.08°C for 47.60 min with an ethanol-water ratio of 24.02%. Organosolv pre-treatment is an effective process for delignification of the lignocellulosic structure.

Demineralized Bone Matrix Fibers plus Allograft Bone for Multilevel Posterolateral Spine Fusion: A Game Changer?

Introduction: While autograft bone is the gold standard for multilevel posterolateral lumbar fusion, bone substitutes and graft extenders such as allograft bone, ceramics and demineralized bone matrix (DBM) have been used to avoid the morbidity and insufficient quantity associated with harvesting autologous bone. The primary objective of this retrospective study was to determine whether, in patients with increased risk of operative nonunion related to multilevel fusion, adding DBM fibers to mineralized bone allograft resulted in better fusion than using allograft alone. The secondary objectives were to evaluate how adding DBM fibers affects functional disability, low back pain, intraoperative blood loss and the nonunion rate. Methods: This retrospective study involved a chart review of consecutive patients who underwent multilevel lumbar spinal fusion and were operated on by a single surgeon. The patients were divided into two groups: 14 patients received mineralized bone allograft (control group) and 14 patients received a combination of mineralized bone allograft and DBM (experimental group). Patients were reviewed at a mean of 16.4 ± 2.2 months after surgery at which point CT scans were analyzed to determine whether fusion had occurred;Oswestry disability index (ODI) and pain were also evaluated. Results: A mean of 5 levels [min 2, max 13] were fused in these patients. Posterolateral fusion as defined by the Lenke classification was not significantly different between groups. The experimental DBM group had a significantly better composite fusion score than the control group (P Discussion: Adding DBM fibers to allograft bone during multilevel posterolateral spinal fusion was safe and produced better composite fusion than using allograft only as an autograft extender.

Characterization and Modeling of Mechanical Properties of Additively Manufactured Coconut Fiber-Reinforced Polypropylene Composites

In the face of the increased global campaign to minimize the emission of greenhouse gases and the need for sustainability in manufacturing, there is a great deal of research focusing on environmentally benign and renewable materials as a substitute for synthetic and petroleum-based products. Natural fiber-reinforced polymeric composites have recently been proposed as a viable alternative to synthetic materials. The current work investigates the suitability of coconut fiber-reinforced polypropylene as a structural material. The coconut fiber-reinforced polypropylene composites were developed. Samples of coconut fiber/polypropylene (PP) composites were prepared using Fused Filament Fabrication (FFF). Tests were then conducted on the mechanical properties of the composites for different proportions of coconut fibers. The results obtained indicate that the composites loaded with 2 wt% exhibited the highest tensile and flexural strength, while the ones loaded with 3 wt% had the highest compression strength. The ultimate tensile and flexural strength at 2 wt% were determined to be 34.13 MPa and 70.47 MPa respectively. The compression strength at 3 wt% was found to be 37.88 MPa. Compared to pure polypropylene, the addition of coconut fibers increased the tensile, flexural, and compression strength of the composite. In the study, an artificial neural network model was proposed to predict the mechanical properties of polymeric composites based on the proportion of fibers. The model was found to predict data with high accuracy.

Improvement of Mechanical Qualities of Clay Material through Coconut Fiber Stabilization

The criticisms regularly formulated towards clay or soil, in general, are its weak mechanical qualities and low water quality. Therefore, it is necessary to find techniques to improve the properties of this material, which is widely used worldwide. Here, we propose stabilizing clay with coconut fiber as a solution to enhance its mechanical properties. To do this, we used an experimental method, first determining the geotechnical properties of the clay and then its mechanical properties. The geotechnical study using the Proctor Test revealed that the dry density of the clay is γb = 1.42 g/cm3, and its water content is W = 22.3%. By applying the rolling method, the Atterberg limits were determined: liquid limit Wl = 63.6, plastic limit Wp = 27.9, plasticity index Ip = 35.7, and consistency index Ic = 1.46. With 25 P = 35.7 1.3, according to the water classification, it falls into class A3ts. The mechanical part focused on compression and flexural strengths obtained using a PROETI hydraulic press. We obtained a flexural strength of 0.63 MPa for simple clay (BA);0.89 MPa for clay + 0.25% fiber (BAF1/4);1.68 MPa for clay + 0.5% fiber (BAF1/2);1.87 MPa for clay + 0.75% fiber (BAF3/4);and 3.91 MPa for clay + 1% fiber (BAF1). As for the compression strength, BA = 5.90 MPa, BAF1/4 = 6.395 MPa, BAF1/2 = 6.292 MPa, BAF3/4 = 6.065 MPa, and BAF1 = 5.423 MPa. The addition of fiber has thus improved the mechanical qualities of the simple clay. These stabilized bricks can be used for sustainable and bioclimatic construction, providing higher durability and good comfort.

Development of Kaolin and Glass Fiber Reinforced Composites for Thermal Insulating Panels

In our modern world, where conserving energy is highly valued, thermal insulation panels play a crucial role in reducing heat transfer between two spaces, surfaces, or materials. They are used to enhance the energy efficiency of various industrial applications by minimizing heat loss and temperature control. These panels function as silent protectors, aiding in reducing energy consumption and making things more sustainable and better for the environment. This is where composite materials come in;they are known for their lightweight nature, high strength-to-weight ratio, and excellent thermal insulation properties and have gained significant attention. Researchers are actively engaged in various studies aimed at enhancing these materials further. This research project focuses on the development of kaolin and glass fiber-reinforced composites for thermally insulating panels, to which natural strengthening materials like corn husk and bamboo fibers are added. The aim is to create cost-effective and efficient composite materials for thermal insulation applications by incorporating these components with a binder consisting of potassium silicate, hydroxide, and distilled water. This project involves conducting compression tests, bending tests, impact tests, thermal conductivity measurements, and microscopic analysis to evaluate the mechanical and thermal properties of the developed composites. The profound impact of these engineered composites on thermal insulation panels stands to revolutionize energy conservation efforts, offering a potent avenue to minimize heat loss and enhance overall energy efficiency across an array of industrial sectors.

Coconut Fiber Pyrolysis: Bio-Oil Characterization for Potential Application as an Alternative Energy Source and Production of Bio-Degradable Plastics

The current energy crisis could be alleviated by enhancing energy generation using the abundant biomass waste resources. Agricultural and forest wastes are the leading organic waste streams that can be transformed into useful alternative energy resources. Pyrolysis is one of the technologies for converting biomass into more valuable products, such as bio-oil, bio-char, and syngas. This work investigated the production of bio-oil through batch pyrolysis technology. A fixed bed pyrolyzer was designed and fabricated for bio-oil production. The major components of the system include a fixed bed reactor, a condenser, and a bio-oil collector. The reactor was heated using a cylindrical biomass external heater. The pyrolysis process was carried out in a reactor at a pressure of 1atm and a varying operating temperature of 150˚C, 250˚C, 350˚C to 450˚C for 120 minutes. The mass of 1kg of coconut fiber was used with particle sizes between 2.36 mm - 4.75 mm. The results show that the higher the temperature, the more volume of bio-oil produced, with the highest yield being 39.2%, at 450˚C with a heating rate of 10˚C/min. The Fourier transformation Infrared (FTIR) Spectroscopy analysis was used to analyze the bio-oil components. The obtained bio-oil has a pH of 2.4, a density of 1019.385 kg/m3, and a calorific value of 17.5 MJ/kg. The analysis also showed the presence of high-oxygenated compounds;carboxylic acids, phenols, alcohols, and branched oxygenated hydrocarbons as the main compounds present in the bio-oil. The results inferred that the liquid product could be bestowed as an alternative resource for polycarbonate material production.

Physical and Thermo-Mechanical Properties of Composite Materials Based on Raw Earth and Crushed Palm Leaf Fibers (Borassus aethiopum)

The objective of this study is to seek solutions to reduce the impact of buildings on climate change and to promote the use of local bio-sourced or geo-sourced materials for sustainable construction. Different samples of raw earth from 3 sites were taken in the commune of Mlomp. Geotechnical tests showed that the raw earth samples from sites 2 and 3 have more clay fraction while site 1 contains more sand. The fact of integrating fibers from crushed palm leaves (Borassus aethiopum) (2%, 4% and 6%) into the 3 raw earth samples reduced the mechanical resistance to compression and traction of the 3 raw earths. The experimental results of thermal tests on samples of earth mixtures with crushed Palma leaf fibers show a decrease in thermal conductivity as well as thermal effusivity as the percentages increase (2%, 4% and 6%) of fibers in raw earth for the 3 sites. This shows that this renewable composite material can help improve the thermal insulation of building envelopes.

Indirect Electroanalysis of 3-Methyl-4-Nitrophenol in Water Using Carbon Fiber Microelectrode Modified with Nickel Tetrasulfonated Phthalocyanine Complex

Electrochemical detection of 3-methyl-4-nitrophenol (MNP) in direct phenol oxidation occurs at high potentials and generally leads to progressive passivation of the electrochemical sensor. This study describes the use of a carbon fiber microelectrode modified with a tetrasulfonated nickel phthalocyanine complex for the detection of MNP at a lower potential than that of direct phenol oxidation. The MNP voltammogram showed the presence of an anodic peak at -0.11 V vs SCE, corresponding to the oxidation of the hydroxylamine group generated after the reduction of the nitro group. The effect of buffer pH on the peak current and SWV parameters such as frequency, scan increment, and pulse amplitude were studied and optimized to have better electrochemical response of the proposed sensor. With these optimal parameters, the calibration curve shows that the peak current varied linearly as a function of MNP concentration, leading to a limit of detection (LoD) of 1.1 μg/L. These results show an appreciable sensitivity of the sensor for detecting the MNP at relatively low potentials, making it possible to avoid passivation phenomena.

Exploring the Synergy: Experimental and Theoretical Investigation of Steel and Glass Fiber Reinforced Polymer (GFRP) Reinforced Slab Incorporating Alccofine and M-Sand

Introduction: This study investigates the Experimental and Theoretical Investigation of Steel and Glass Fiber Reinforced Polymer (GFRP) Reinforced Slab Incorporating Alccofine and M-sand. Objective: Specific objectives include evaluating the mechanical properties and structural behaviour of steel and GFRP-reinforced one-way slabs and comparing experimental and theoretical predictions. Methods: Four different mix proportions were arrived at, comprising both conventional concrete and Alccofine-based concrete. In each formulation, a combination of normal river sand and M-sand was utilized. Results: Concrete with Alccofine exhibits superior mechanical properties, while M-sand incorporation minimally affects strength but reduces reliance on natural sand. GFRP-reinforced slabs display distinct brittle behaviour with significant deflections post-cracking, contrasting steel-reinforced slabs’ gradual, ductile failure. Discrepancies between experimental data and design recommendations underscore the need for guideline refinement. Conclusion: Alccofine and M-sand enhance concrete properties, but reinforcement type significantly influences slab behaviour. GFRP-reinforced slabs, though exhibiting lower values than steel, offer advantages in harsh environments, warranting further optimization.

Study of the Diffusion Behavior of Seawater Absorption in Multi-Walled Carbon Nanotubes/Halloysite Nanotubes Hybrid Nanofillers Modified Epoxy-Based Glass/Carbon Fiber Composites

In the maritime industry, cost-effective and lightweight Fiber Reinforced Polymer (FRP) composites offer excellent mechanical properties, design flexibility, and corrosion resistance. However, their reliability in harsh seawater conditions is a concern. Researchers address this by exploring three approaches: coating fiber surfaces, hybridizing fibers and matrices with or without nanofillers, and interply rearrangement. This study focuses on evaluating the synergistic effects of interply rearrangement of glass/carbon fibers and hybrid nanofillers, specifically Multi-walled carbon nanotubes (MWCNT) and Halloysite nanotubes (HNT). The aim is to enhance impact properties by minimizing moisture absorption. Hybrid nanocomposites with equal-weight proportions of two nanofillers: 0 wt.%, 1 wt.%, and 2 wt.% were exposed to seawater for 90 days. Experimental data was subjected to modelling through the application of Predictive Fick’s Law. The study found that the hybrid composite containing 2 wt.% hybrid nanofillers exhibited a 22.10% increase in impact performance compared to non-modified counterparts. After 90 days of seawater aging, the material exhibited enhanced resistance to moisture absorption (15.74%) and minimal reduction in impact strength (8.52%) compared to its dry strength, with lower diffusion coefficients.

Influence of Waveguide Properties on Wave Prototypes Likely to Accompany the Dynamics of Four-Wave Mixing in Optical Fibers

In this article, we study the impacts of nonlinearity and dispersion on signals likely to propagate in the context of the dynamics of four-wave mixing. Thus, we use an indirect resolution technique based on the use of the iB-function to first decouple the nonlinear partial differential equations that govern the propagation dynamics in this case, and subsequently solve them to propose some prototype solutions. These analytical solutions have been obtained;we check the impact of nonlinearity and dispersion. The interest of this work lies not only in the resolution of the partial differential equations that govern the dynamics of wave propagation in this case since these equations not at all easy to integrate analytically and their analytical solutions are very rare, in other words, we propose analytically the solutions of the nonlinear coupled partial differential equations which govern the dynamics of four-wave mixing in optical fibers. Beyond the physical interest of this work, there is also an appreciable mathematical interest.

鸭腺病毒3型Fiber-2蛋白单克隆抗体的制备及初步应用

为进一步研究鸭腺病毒3型(DAdV-3)Fiber-2蛋白,研究通过PCR扩增fiber-2基因,构建p ET32a-DAdV3-fiber2原核质粒,表达并纯化蛋白,以纯化的蛋白为免疫原,免疫6周龄雌性BALB/c小鼠制备抗Fiber-2单克隆抗体杂交瘤细胞,采用间接免疫荧光试验和免疫印迹试验进行鉴定,并用于DAdV-3 Fiber-2杆状病毒表达产物的鉴定。结果显示:原核表达获得1.77 mg/mL Fiber-2蛋白,筛选获得2株阳性单克隆细胞株2E10E4和4F7F10,两株单克隆抗体均为IgG 1型,轻链类型为κ;DAdV-3感染的LMH细胞中存在特异性荧光,免疫印迹试验确认在约59 ku处出现特异性条带;采用制备的单克隆抗体确认了表达Fiber-2蛋白的重组杆状病毒。研究获得了针对DAdV-3 Fiber-2蛋白的单克隆抗体,为深入探究Fiber-2在DAdV-3感染机制中的作用以及为建立DAdV-3抗原免疫测定技术奠定了基础。

3D-SiO_(2)-fiber中间层对SiC与Nb真空钎焊的影响

缓解异种材料钎焊接头残余应力过程中,针对颗粒增强相添加量少、团聚等问题,引入一种新型三维SiO_(2)短纤维编织,呈疏松、多孔结构的复合材料中间层(3D-SiO_(2)-fiber)辅助SiC陶瓷与Nb钎焊连接.采用SEM,XRD和电子万能试验机等对活性钎料在3D-SiO_(2)-fiber中间层表面润湿性、接头微观组织和力学性能进行表征与分析.结果表明,当Ti元素含量从4.5%增加到6.0%,AgCu-Ti活性钎料在3D-SiO_(2)-fiber中间层表面润湿角从90°降低到3°,润湿性得到显著改善,且AgCu-Ti活性钎料浸入3D-SiO_(2)-fiber中间层深度不断增加;随着钎焊温度的提高(950~980℃)及保温时间(10~25 min)的延长,SiO_(2)短纤维与AgCu-6.0Ti活性钎料反应逐渐充分.该方法不仅保证SiO_(2)短纤维能够大量添加到合金钎料中,而且形成Cu_(3)Si,TiSi,α-Ti及Ti_(2)Cu颗粒相弥散分布于焊缝中,从而有效缓解钎焊接头残余应力,提高接头力学性能,实现陶瓷与金属复合构件的可靠连接.

牛腺病毒3型感染状况及其fiber轴区基因检测

本研究旨在通过TB Green Real-time PCR的方法对国内6个省(四川省、山西省、河南省、河北省、江苏省和内蒙古自治区)的9个规模化养牛场140份以流鼻涕、发热、咳嗽、呼吸困难为特征的病牛呼吸道鼻拭子样本开展牛腺病毒3型(bovine adenovirus type 3,BAdV-3)感染状况调查,并对其中的阳性样本进行fiber轴区基因检测。结果显示,140份样品中BAdV-3阳性率为36.4%,除河北省以外,其余各省均有BAdV-3检出,其中,四川及河南省检出率最高,分别达到82.9%(29/35)和83.3%(10/12)。其中,传统型占总阳性比的80.4%,fiber轴区缺失毒株占总阳性样本的19.6%。缺失位置与数量一致,轴区连续缺失237个碱基对(79个氨基酸),且缺失型仅在四川和河南两省检出,检出率分别为24.1%(7/29)和30%(3/10),场群阳性率分别为100%(2/2)和100%(1/1)。结果表明,BAdV-3已在我国牛场中广泛分布,目前流行的主要以传统型毒株为主,fiber轴区缺失毒株在我国具有独特的地域分布,主要在四川和河南省流行。

禽腺病毒4型fiber-2与8b型fiber截短融合蛋白的原核表达及其免疫原性分析

利用原核表达系统表达Ⅰ群禽腺病毒血清4型(FAdV-4)fiber2和8b型(FAdV-8b)fiber全长蛋白、截短蛋白及二价融合蛋白,并评价其免疫原性。基于FAdV-4 fiber2和FAdV-8b fiber抗原性好的截短片段(FP和BP),利用SOE-PCR构建二价融合片段(BP-FP),分别对截短蛋白、融合蛋白及全长(FL和BL)在大肠杆菌中进行表达,采用SDS-PAGE和Western blot对重组蛋白进行鉴定。纯化后的重组蛋白免疫SPF鸡并攻毒,计算各组存活率、间接ELISA检测免疫鸡血清抗体水平、qPCR检测泄殖腔拭子排毒量和肝脏中细胞因子IL-4、IFN-γ和TNF-αmRNA表达水平以及进行组织病理观察。结果显示:构建的重组质粒均在大肠杆菌中成功可溶表达;BP-FP4组存活率为58.3%,其他组存活率均为100%;全长组(FL组和BL组)抗体水平均高于各自片段组(FP组和BP组),但差异不显著(P>0.05);攻毒后3和7 d C8b组排毒量均显著高于免疫组(P<0.05),攻毒后7 d BP-FP8b组排毒量显著高于BP和BL组(P<0.05),BL组和BP组差异不显著(P>0.05);免疫组IL-4、IFN-γ和TNF-αmRNA表达均显著高于阴性对照组(P<0.05),FL组IFN-γ和TNF-αmRNA表达均显著低于BP-FP4组(P<0.05),FL组和FP组差异不显著(P>0.05),BL组IL-4和IFN-γmRNA表达均显著低于BP-FP8b组(P<0.05)。组织病理观察发现,攻毒组肝脏病变最严重,FP组与BP组次之,FL组与BL组无明显病变。本研究表明FAdV-4 fiber2和FAdV-8b fiber的全长蛋白和截短蛋白免疫保护效果优于融合蛋白,截短蛋白可替代全长蛋白用于制备禽腺病毒亚单位疫苗。

禽腺病毒血清4型和8b型串联截短Fiber2-Fiber蛋白在毕赤酵母中的优化表达

【目的】在毕赤酵母中构建禽腺病毒(FAdV)血清4型和8b型串联截短Fiber2-Fiber蛋白表达系统,并探索适宜的表达条件。【方法】以前期构建的重组质粒pCold-Fiber2-Fiber为模板,用PCR法扩增Fiber2-Fiber片段,连接pPICZαA载体。经双酶切和测序鉴定后的阳性重组质粒pPICZαA-Fiber2-Fiber电转化毕赤酵母GS115。经MD平板和不同浓度Zeocin抗性筛选后进行PCR扩增和测序鉴定得到阳性菌株;用甲醇诱导表达阳性菌株,72 h后收集蛋白上清进行SDS-PAGE和液相色谱串联质谱(LC-MS/MS)鉴定。采用单因素变量法对不同pH(5.0、6.0、7.0和8.0)、甲醇浓度(0.5%、1.0%、1.5%和2.0%)、时间(24、48、72和96 h)进行表达条件优化,用SDS-PAGE鉴定,并用间接ELISA和Western blotting验证重组蛋白的反应原性。【结果】成功构建了重组质粒pPICZαA-Fiber2-Fiber,对表达72 h后的上清进行SDS-PAGE发现1条约70 ku的特异条带,经LC-MS/MS鉴定为目的蛋白。串联截短Fiber2-Fiber蛋白的最优诱导条件为pH 6.0、1%甲醇、诱导时间72 h。间接ELISA和Western blotting结果显示该重组蛋白能与FAdV-4和FAdV-8b阳性血清特异性结合。【结论】Fiber2-Fiber串联截短蛋白可在毕赤酵母中稳定表达并具有良好的反应原性,为禽腺病毒双价亚单位疫苗的研发奠定基础。

Ⅰ群禽腺病毒(4型)Fiber-2蛋白亚单位疫苗与全病毒灭活疫苗的免疫效果比较

为了比较不同抗原含量的Ⅰ群禽腺病毒(4型)Fiber-2蛋白亚单位疫苗与全病毒灭活疫苗的免疫效力差异,本研究制备不同抗原含量的亚单位疫苗和全病毒灭活疫苗,采用超低免疫剂量20μL/只免疫SPF鸡,免疫21 d后攻毒,通过死亡率、临床症状、大体剖检病变、免疫组化和泄殖腔拭子排毒等进行免疫效力比较。试验结果表明,亚单位疫苗抗原含量AGP效价不低于1∶8,全病毒灭活疫苗抗原含量为10^(7.0)TCID_(50)/0.1 mL时,可抵抗Ⅰ群禽腺病毒(4型)强毒株的攻击,提供100%的免疫攻毒保护,攻毒后3、5、7 d泄殖腔拭子均未见排毒,且免疫组化均为阴性。亚单位疫苗抗原含量AGP效价为1∶4,全病毒灭活疫苗抗原含量为106.5 TCID_(50)/0.1 mL时,在超低免疫剂量情况下,仍可提供至少80%的免疫攻毒保护,且攻毒后3、5、7 d泄殖腔拭子排毒率仅为2/10~3/10,免疫组化阳性率仅为1/10。但全病毒灭活疫苗抗原含量为106.0 TCID_(50)/0.1 mL时,免疫攻毒保护效果不理想。综合以上结果表明,Fiber-2蛋白具有良好的免疫原性,抗原含量AGP效价不低于1∶4,与全病毒灭活疫苗抗原含量为106.5 TCID_(50)/0.1 mL免疫效力相当,可用于疫苗研制开发。