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研究灌胃猪血浆IgG提取物对小白鼠免疫功能的影响。采用腹腔注射环磷酰胺制造试验小白鼠的免疫功能抑制模型,选用齐墩果酸为阳性对照组,猪血浆IgG提取物为试验组,预饲5d后,然后将猪血浆IgG提取物按每千克体重45、90和180mg3种剂量给小白鼠连续灌胃10d后测定小白鼠免疫水平的变化。小白鼠灌胃猪血浆IgG提取物后,对抗由环磷酰胺引起的小白鼠免疫器官萎缩有一定的保护作用,并对环磷酰胺所致动物DTH反应低下有一定的对抗作用;能显著增加小白鼠血清中IgG的含量和免疫球蛋白的含量(P<0.05),能显著增加小白鼠血清中球蛋白的含量(P<0.05),灌胃猪血浆IgG提取物能显著提高小白鼠的免疫功能。 相似文献
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许多免疫学试验(如ELISA、荧光抗体技术)都要应用提纯的IgG。传统的提取IgG的方法是用Na_2SO_4沉淀血清,然后再进行离子交换层析或凝胶电泳。以下方法具有简单易行、费用低,可同时测定多个样品等优点,且提取的IgG纯度不比市售的IgG低。现介绍如下: 采取鸡的血液,凝血析出血清。取此血清10ml,加入4倍量的醋酸盐缓冲液(60mM、pH4.0),用1N NaOH调整pH值为4.5。然后边搅拌边加入辛酸,至血清稀释液中辛酸的含量达25μl/ml为止。将此溶液继续搅拌30分钟(以上操作均在室温时进行,以下操作须 相似文献
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本试验旨在研究新生羔羊血清免疫球蛋白和乳铁蛋白（LF）含量随时间变化的规律，并验证白利度（Brix）法测定新生羔羊被动免疫状态的可行性。选择8只初生重（3.53±0.33）kg、性别一致（公羊）的健康随母哺乳湖羊羔羊，采集出生后0、6、12、24、36 h和72 h血清样品，利用单向免疫扩散法（RID）测定免疫球蛋白G(IgG)含量，酶联免疫吸附测定法（ELISA）测定免疫球蛋白A(IgA)、免疫球蛋白M(IgM)和LF含量，同时利用手持折光仪和数显折光仪（Brix法）检测血清IgG含量，评估白利度法评估血清IgG含量的准确性。结果显示：IgG含量随产后时间延长呈先升高后降低的二次曲线变化（P<0.05）;IgA和IgM含量整体稳定；LF在0 h时血清含量为0 mg/mL,6 h后整体含量稳定在0.05 mg/mL左右。使用手持折光仪读数预测羔羊血清IgG含量公式为：Y=16.79×X–58.55(R2=0.61,P<0.05)；使用数显折光仪读数预测羔羊血清IgG含量公式为：Y=14.50×X–49.38(R2=0.49,P<0.05)。采用血清IgG含量高于25 ... 相似文献
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为了探究重链抗体在骆驼免疫保护中的生物学作用,本研究利用Protein G和Protein A亲和层析纯化新疆双峰驼血清中总IgG和重链抗体IgG2,并免疫昆明小白鼠制备抗双峰驼总IgG和抗双峰驼重链抗体IgG2的多抗血清;通过ELISA检测双峰驼在体液免疫应答过程中针对spaA-N、溶菌酶和蒜氨酸酶3种抗原的重链抗体的滴度变化。结果从新疆双峰驼血清中亲和层析纯化出天然重链抗体IgG2,蛋白质分子质量约为46 ku;免疫昆明小白鼠后获得抗双峰驼总IgG多抗血清的效价为1∶204800,抗双峰驼重链抗体IgG2多抗血清的效价为1∶6400。在spaA-N免疫骆驼诱导的体液免疫应答过程中,重链抗体IgG2出现延迟反应。3种蛋白均能诱导抗原特异的IgG2亚类重链抗体的产生。 相似文献
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本试验采用辛酸去脂、饱和硫酸铵溶液盐析法提纯鹅卵黄抗体IgY,采用SDS-PAGE方法分析其纯度;用纯化后的IgY免疫试验兔,收集血清,测定血清抗体效价;采用辛酸-硫酸铵法结合Protein G树脂亲和层析法纯化免血清IgG,用改良的高碘酸钠法进行辣根过氧化物酶标记,直接ELISA法测定标记物效价.结果表明,提纯的鹅卵黄IgY浓度为11.5 mg/mL,纯度约为92％;血清琼脂扩散抗体效价为1∶32;Western Blotting试验证明,兔血清中含有抗鹅卵黄抗体IgY重链及轻链的特异性抗体;直接ELISA法测得标记物效价为1:1 100. 相似文献
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为了研究美洲大蠊药物残渣对小白鼠生长性能、脏器指数、血清免疫球蛋白含量的影响,试验采用单因素完全随机设计,将32只体重为18～22 g的SPF级昆明种雄性小白鼠随机分为4组,每组8只。对照组饲喂基础日粮,试验组(0.5%、1%、2%剂量组)分别饲喂在基础日粮中添加0.5%、1%、2%美洲大蠊药物残渣粉的试验日粮,试验期30 d。试验测定美洲大蠊药物残渣的基础营养成分,包括粗蛋白、粗脂肪、水分及粗灰分,试验结束后测定小白鼠的体重、胸腺指数、脾脏指数及血清免疫球蛋白(IgG、IgM、IgA)含量。结果表明：美洲大蠊药物残渣中粗蛋白和粗脂肪含量分别为64.93%和29.58%,水分含量为3.71%,粗灰分含量为3.05%。与对照组相比,各试验组小白鼠的体重、脏器指数均有提高,但差异不显著(P>0.05)。1%、2%剂量组小白鼠的血清IgG、IgM、IgA含量极显著高于对照组(P<0.01)。说明美洲大蠊药物残渣能在一定程度上提高小白鼠的免疫功能和生长性能。 相似文献
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为建立大熊猫IgG的定量双抗体夹心ELISA检测方法(DAS-ELISA),本研究纯化了大熊猫血清中IgG,并将其作为免疫原分别免疫家兔和豚鼠,制备兔抗和豚鼠抗大熊猫IgG高免血清,经优化DAS-ELISA反应条件,建立了以纯化的兔抗大熊猫IgG作为捕获抗体和豚鼠抗大熊猫IgG高免血清作为检测大熊猫IgG的定量DAS-ELISA 检测方法.该方法能检测1.5625μg/mL～100 μg/mL的大熊猫IgG,大熊猫IgG含量(Y)与DAS-ELISA检测OD值(X)之间呈良好的线性关系,Y=4.7186X+0.044,相关系数为0.9911,该方法特异性强,稳定性好,能用于大熊猫IgG的定量检测. 相似文献
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共轭亚油酸对小白鼠体脂沉积及免疫功能的影响 总被引：1，自引：0，他引：1
试验研究了日粮中添加共轭亚油酸(CLA)对小白鼠脂肪沉积与免疫的影响。试验选择体重相近的4周龄的雄性昆明种小白鼠40只,随机分成4组,每组10只,在日粮中分别添加0%、0.1%、0.5%、1.0%的CLA,喂养4周后,测定小白鼠腹脂重、体脂含量、脂肪细胞大小与数目、免疫器官重量与指数、T淋巴细胞转化率、血清IgG、IgM与IgA。结果显示,当日粮中CLA的添加量为0.5%时,可以显著提高小白鼠脾脏重量与指数、T淋巴细胞转化率及IgM含量,降低小白鼠体重、腹脂重、体脂含量与脂肪细胞直径(P<0.05)。CLA的添加量为1.0%时,可以显著提高小白鼠IgG含量(P<0.05)。而添加CLA对脂肪细胞数目与IgA影响不显著(P>0.05)。说明饲喂一定量的共轭亚油酸可减少小白鼠体脂沉积,提高免疫力。 相似文献
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分别采用饱和硫酸铵法和辛酸—硫酸铵法纯化兔抗酸性红73(acid red 73,AR73)免疫球蛋白G(IgG)。通过二喹啉甲酸(BCA)法、十二烷基硫酸钠—聚丙烯酰胺凝胶电泳(SDS-PAGE)试验、间接ELISA法分别测定IgG的浓度、纯度及效价。结果显示,饱和硫酸铵法所提IgG浓度为36.44 mg/mL,最高效价达1.6×105;辛酸—硫酸铵法所提IgG浓度为15.79 mg/mL,最高效价达3.2×105。提示,辛酸—硫酸铵法IgG纯度优于饱和硫酸铵法。 相似文献
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G E Rumbaugh A A Ardans D Ginno A Trommershausen-Smith 《Journal of the American Veterinary Medical Association》1978,172(3):321-325
Four procedures for assessment of adequacy of colostral immunoglobulin (Ig) transfer in foals were evaluated. Results of zinc sulfate turbidity test, serum electrophoresis, total serum protein refractometry, and sodium sulfite precipitation test were compared with immunoglobulin G content determined by single radial immunodiffusion. The zinc sulfate turbidity test gave acceptable results for IgG, except that hemolyzed serum samples gave higher than expected values. A correction factor for hemolyzed serum was found to be useful. Serum electrophoresis was a satisfactory method of estimating IgG content. Total serum protein values may not be a valid basis for estimating IgG content, inasmuch as postsuckling total protein values were found to decrease in some foals in which passive transfer of IgG had been adequate. Sodium sulfite precipitation reactions were too unpredictable to be of value for determination of neonatal IgG concentration. 相似文献
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J.G. Mohanty Y. Elazhary 《Comparative immunology, microbiology and infectious diseases》1989,12(4):153-160
Immunoglobulin G (IgG) from bovine serum raised against Aeromonas Salmonicida was purified by ammonium sulphate precipitation (ASP) or caprylic acid treatment followed by ammonium sulphate precipitation (CAAS). Purity of IgG samples prepared by both methods were examined by High Performance Gel Permeation Chromatography, electrophoresis and antibody activity assay. Results suggest that IgG prepared by ASP is better than that obtained by CAAS method in terms of the yield of the IgG monomers and the recovery of the antibody activity. 相似文献
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Immunoturbidimetric determination of serum IgG concentration in foals was compared with the reference methods of single radial immunodiffusion and serum protein electrophoresis. High positive correlations were discovered when the technique was compared with either of these reference methods. The zinc sulfate turbidity test for serum IgG estimation was also evaluated. Although a positive correlation was discovered when the latter method was compared with reference methods, it was not as strong as the correlation between reference methods and the immunoturbidimetric method. The immunoturbidimetric method used in this study is specific and precise for equine serum IgG determination. It is rapid and, thus, is advantageous when timely evaluation of critically ill foals is necessary. The technique should be adaptable to various spectrophotometers and microcomputers for widespread application in veterinary medicine. 相似文献
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Harada E Araki Y Furumura E Takeuchi T Sitizyo K Yajima T Kuwata T 《Journal of veterinary medicine. A, Physiology, pathology, clinical medicine》2002,49(7):358-364
The characteristic transfer of colostral components into cerebrospinal fluid (CSF) via serum after natural suckling has been studied by sodium dodecyl sulphate (SDS) electrophoresis, two-dimensional electrophoresis, immunoblot and enzyme linked immunosorbent assay methods in non-suckling pigs. Total protein concentrations in the serum increased immediately after first suckling, reached a peak value at 12 h, corresponding to a 2.3-fold compared with pre-suckling level. The protein concentration in CSF also increased and reached peak value at 6 h corresponding to 1.6-fold compared with presuckling level. IgG in serum not detected before suckling, increased steeply after suckling, IgG, IgM and IgA transported into the serum were observed in completely intact form by immunoblot method. The IgG transported into serum was quickly transferred into CSF after natural suckling in contrast to the case of bovine IgG. Serum concentration of transferrin was maintained at high level before suckling and was not changed by suckling. Transferrin also detected in CSF was not changed by suckling. Bovine lactoferrin (Lf) administered into the intestinal lumen was transported into serum (0.01%) and also detected in CSF after 6 h as undegraded form (3.1%). Thus, homologous IgG and bovine Lf are transported into CSF, suggesting that the transport of macromolecules into CSF is selective in neonatal pigs. 相似文献
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A method for screening of immunoglobulin levels in porcine fetuses is presented. The method is based on rocket electrophoresis. This method is sensitive, but when used for assaying immunoglobulins it is only semiquantitative. It was shown that sera from 11 hysterectomy-derived colostrum-deprived neonatal pigs from a normal gilt contained no IgM or IgA, but small amounts of IgG. Sera from 9 piglets isolated in the same way from a gilt infected with porcine parvovirus contained all 3 antibody classes. IgM was detected by an interspecies reaction using anti human μ-chain immunoglobulin. 相似文献
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Christine S. Olver Teckla L. Webb Lindsey J. Long Hataichanok Scherman Jessica E. Prenni 《Veterinary clinical pathology / American Society for Veterinary Clinical Pathology》2010,39(3):337-345
Background: Disease‐specific biomarkers hold diagnostic promise in both human and veterinary medicine, but serum biomarkers in low concentrations may be masked by the presence of abundant proteins, mostly albumin and IgG. Methods to deplete albumin and IgG exist, but efficacy of these methods for depleting equine serum of these proteins has not been established. Objective: The aim of this study was to determine if albumin and IgG could be depleted from equine serum using several commercially available kits and procedures. Methods: One‐dimensional gel electrophoresis followed by densitometry was used to determine percent of albumin, IgG, and both in pooled serum from 3 horses before and after application of 7 depletion methods. Repeatability was determined by applying the 2 best methods to serum samples from 6 grade horses. Results: For pooled serum, depletion rates varied from 35–90% for albumin and 0–94% for IgG. In the repeatability study, the ProteoExtract method combined with protein G Sepharose beads to remove additional IgG provided the best overall performance with 66% albumin depletion and 100% IgG depletion. A protocol using protein G Sepharose beads to remove IgG followed by ethanol precipitation of nonalbumin proteins with albumin remaining in the supernatant was the second most effective, with 85% albumin depletion and 55% IgG depletion. Although a multiprotein immunodepletion column effectively removed 90% of the albumin, the method was ineffective at removing IgG. Conclusion: Albumin and IgG removal kits optimized for human use have variable efficacy for equine serum. Combined use of the ProteoExtract kit and manual incubation with protein G Sepharose beads provided the most effective depletion. 相似文献
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两种方法纯化免抗绵羊肺炎支原体IgG的比较 总被引：2，自引：0，他引：2
目的:比较两种方法纯化兔抗绵羊肺炎支原体IgG的效果。方法:采用饱和硫酸铵和辛酸-硫酸铵两种方法分离纯化兔抗绵羊肺炎支原体抗体,并用十二烷基硫酸钠聚丙烯酰胺凝胶电泳(SDS-PAGE)、分光光度计和琼脂扩散(AGP)试验对纯化产物进行相对分子质量、蛋白质含量及免疫活性进行鉴定。结果:纯化后抗体的蛋白含量基本相同;产物纯度和效价以辛酸-硫酸铵法较高。结论:辛酸硫酸铵法是一种简便、快速、高效的抗体纯化方法。 相似文献