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采用饱和硫酸铵分步盐析法结合柱层析提取，纯化欧洲鳗血清免疫球蛋白（Ｉg) ,实验证实欧洲鳗Ｉg主要分布在硫酸铵饱和度３０％－５０％的区间内，Ｓephacry1-S200进一步提纯的Ｉg主要存在于第一个蛋白峰，而Ｓepharose-4B柱层析提纯的Ｉg则存在于第二个蛋白峰，ＤＥＡE-52脑子交换柱层析可进一步纯化Ｓepharose-4B柱层析的产物，ＥＬＩＳＡ和Western-blot分别证实上述提取物具有抗体的免疫学活性，ＳＤＳ－ＰＡＥＧ分析纯化的Ｉg，显示了洲鳗Ｉg重链约为６８kD，轻链约为２６kD. 相似文献
3.
欧洲鳗免疫球蛋白单克隆抗体的制备与特性 总被引：19，自引：2，他引：19
应用杂交瘤单克隆抗体技术制备了13个分泌抗欧洲鳗免疫球蛋白的单克隆抗体细胞株，并对这些单克隆抗体的特性进行了分析。经抗体级份和亚级份测定，其中IgM有3株，IgG1有5株，IgG2a有3株，IgG2b有2株；所有的单克隆抗体与欧鳗免疫球蛋白均有ELISA反应特性，抗体滴度10^4-10^7，有七株单抗具有WESTERN BLOT反应特性，能在变性条件下识别欧鳗免疫球蛋白的重链。进一步实验证实这些单抗能特异地识别欧洲鳗、日本鳗的免疫球蛋白，而与鲫、淡水白鲳、罗非鱼、胡子鲶、美洲斑点叉尾Hui血清免疫球蛋白以及水产动物常见病原菌如气单胞菌、爱德华氏菌、弧菌、柱状屈桡杆菌、沙门氏菌及大肠杆菌等无任何交叉反应。单克隆抗体9D7，对纯化的欧洲鳗免疫球蛋白的检测灵敏度为16ng。实验结果证明这些单抗具有高度特异、高度灵敏等特点，可用于鳗鲡免疫球蛋白的结构分析、免疫应答水平监测和病原诊断，具有广阔的应用前景。 相似文献
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采用饱和硫酸氨分步沉淀和Sephadex G200凝胶层析的方法，首次分别纯化制备了健康非免疫状态下中华鲟（Acipenser sinensis）、史氏鲟（Acipenser schrenckii）和达氏鳇（Huso dauricus）的血清免疫球蛋白（Ig）
，并采用聚丙烯酰胺凝胶电泳（PAGE）、SDS-聚丙烯酰胺凝胶电泳（SDSPAGE）、蛋白免
疫印迹（Western blotting）及免疫琼扩实验等方法对其Ig及Ig亚单位的分子量和部分特性进行了分析。PAGE及SDSPAGE的结果显示：史氏鲟，中华鲟和达氏鳇IgM的相对分子量分别为867 kD， 896 kD和924 kD；3种鲟鱼Ig的重链分子量均为88 kD，都具有29 kD的轻链，其中达氏鳇还另有一分子量约为26 kD的轻链蛋白。分子量的测定及计算结果显示鲟鱼的Ig为四聚体。Western-blotting的检测结果表明，3种鲟鱼Ig的重链与其Ig具有同样的抗原性，在硝酸纤维素膜上可被兔抗鲟Ig多克隆抗体所识别，而轻链的Western blotting检测结果则呈阴性。免疫沉淀反应的结果显示，3种鲟鱼的血清及其Ig与相互之间的兔抗Ig血清有免疫沉淀反应，但与兔抗鲤Ig血清无免疫沉淀反应，这表明3种鲟科鱼类的Ig在结构和序列上是较为相似的，而与鲤鱼等高等硬骨鱼类的Ig存在较大的差别。 相似文献
5.
欧洲鳗鲡血清免疫球蛋白纯化及其结构分析 总被引：8，自引：0，他引：8
应用亲和层析技术提纯欧洲鳗鲡免疫球蛋白（Ig），并对欧鳗Ig的结构和抗原性进行分析。层析结果表明：Ig蛋白呈现一个锐形曲线，蛋白峰与抗原活性峰高度重叠。SDSPAGE分析表明：欧鳗Ig重链分子量为68 kD，轻链有3条，分子量分别为21 kD、23 kD和26 kD。凝胶电泳结果显示：在非变性非还原条件下，欧鳗Ig有790 kD和350 kD 2条蛋白带，在变性非还原条件下有790 kD、593 kD和350 kD 3条蛋白带。Western-blotting试验证实：兔抗欧鳗Ig能识别欧鳗Ig的多种不同聚合体和Ig的重链，但不能识别Ig的轻链。结论：欧鳗Ig在自然条件下可能以四聚体和二聚体的形式存在，这与其它硬骨鱼的Ig形式有差异。欧鳗Ig链间二硫键不健全,在SDS作用下可解聚产生多种不同分子量的聚合体，首次揭示欧鳗Ig的轻链有3种异型。 相似文献
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Tomomasa Matsuyama Chihaya Nakayasu Takamitsu Sakai Norihisa Oseko 《Fisheries Science》2009,75(2):335-341
Two monoclonal antibodies (MAbs: JFW1 and JFW10) were produced against peripheral blood leukocytes (PBL) in Japanese flounder.
Additionally, MAbs against flounder immunoglobulin (Ig; JFW20 and JFW21) were generated for the surface marker of Ig+ leukocytes using purified serum Ig as an antigen. MAb JFW1 recognized the surface marker of granulocytes and monocytes and
MAb JFW10 specifically bound to the surface antigen of thrombocytes. Flow cytometric analysis of PBL incubated with JFW1,
JFW10, JFW20 and JFW21 revealed that 2.5–7.4, 23.7–50.1, 25.2–26.1 and 5.2–8.3% of all leukocytes were positive for these
markers. Analysis of head kidney leukocytes (HKL) showed that JFW1, JFW10, JFW20 and JFW21 bound to 30.5–36.3, 1.9–2.8, 6.4–8.3
and 1.9–3.0% of all leukocytes, respectively. Western blot analysis after SDS-PAGE showed that JFW10 recognizes a protein
of 115 kDa from lysed PBL. JFW20 recognized the 70 and 74 kDa proteins of the heavy chain of Ig from serum. No band was observed
for either JFW1 or JFW21. These antibodies will be useful for the identification and isolation of Japanese flounder leukocyte
subpopulations and will facilitate immunological studies of flounder. 相似文献
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White spot disease (WSD) is an important viral disease of penaeid shrimp caused by white spot syndrome virus (WSSV). WSSV isolated from WSD outbreaks in commercial shrimp (Penaeus monodon) farms in India were propagated in the laboratory in healthy shrimp. The virus was purified from the infected tissues by sucrose gradient centrifugation. The VP28 was electroeluted from SDS-PAGE gels and was used to immunize Balb/c mice to produce hybridomas secreting monoclonal antibodies (MAb) against WSSV. A total of five hybridoma clones secreting MAbs to VP28 were produced. The MAbs were of the isotypes IgG1, IgG2b and IgM. The MAbs reacted with VP28 of WSSV and not with any other viral or shrimp protein in western blot. The MAbs were used to develop dot immunoblot assay using an immunocomb to detect WSSV from field samples. The test developed had an analytical sensitivity of 625 pg and a diagnostic sensitivity of 100% compared to single step polymerase chain reaction (PCR). The test can be used as an alternate for first step PCR to detect WSSV from field samples. 相似文献
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选取4种不同品系罗非鱼, 分别为吉富罗非鱼(Oreochromis niloticus)、奥尼罗非鱼(O. niloticus×O. aureus)、红罗非鱼(O. nilotica♂× O. mossambica♀)和奥利亚罗非鱼(O. aureaus)。在26℃水温下饲养3周后, 选取规格基本一致的罗非鱼(体质量50.73 g±4.23 g)进行低温驯化实验。水温以3℃/d的速度从26℃降至8℃, 分别于水温为26℃、20℃、14℃和8℃时进行采样, 比较不同驯化阶段4种不同品系罗非鱼血清皮质醇和免疫相关指标的变化规律。结果表明, 与26℃时的免疫指标相比, 水温降至8℃时, 吉富与红罗非鱼血清皮质醇水平显著升高; 然而血清C3、C4、IgM以及头肾C型溶菌酶mRNA水平显著下降(P<0.05), 血清中高皮质醇水平对鱼体的免疫产生了抑制作用。水温为8℃时, 奥尼罗非鱼血清皮质醇、IgM和补体C3以及头肾抗菌肽mRNA水平显著升高; 奥利亚血清皮质醇、C4和IgM以及头肾C型溶菌酶mRNA水平与26℃时相比无显著差异, 然而血清溶菌酶与C3水平降低。驯化实验结束后, 比较了4种不同品系罗非鱼在8℃水温下48 h内的累积死亡率。吉富与红罗非鱼组累积死亡率较高, 分别达到43.3%和40.0%; 奥尼罗非鱼其次, 为23.3%; 奥利亚罗非鱼最低, 为20.0%。较高的血清IgM和头肾溶菌酶和抗菌肽mRNA水平可能有助于提高奥尼和奥利亚罗非鱼的抗低温应激能力, 增加低温时的成活率。本研究通过分析4种品系罗非鱼不同驯化阶段血清皮质醇和免疫相关指标, 探讨不同品系罗非鱼在低温驯化过程中的免疫保护机制, 旨在为下一步抗低温新品系罗非鱼的选育提供理论依据。
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Characterization and production of polyclonal antisera against pangasius (Pangasianodon hypophthalmus) serum immunoglobulin IgM derived from DEAE cellulose based ion exchange chromatography 
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下载免费PDF全文 Sudhagar Arun Sudhagar Kurcheti Pani Prasad Marrappan Makesh Govindarajan Rathi Bhuvaneswari Kezhedath Jeena 《Aquaculture Research》2015,46(6):1417-1425
Pangasius (Pangasianodon hypophthalmus) is a commercially important candidate species in freshwater aquaculture and it is important to understand the immune system of pangasius against infectious disease. The present study was aimed at the purification, characterization and quantification of serum IgM in pangasius (P. hypophthalmus). Serum IgM was purified by diethylaminoethyl (DEAE) cellulose based ion exchange chromatography. The molecular weight of native immunoglobulin was found to be 798 kDa. Heavy (H) and light chains were found to possess molecular weight of 70.1 and 26 kDa respectively. A 248 bp segment of IgM H chain gene of pangasius was amplified and sequenced (partial). The antisera raised against pangasius immunoglobulin cross‐reacted with the immunoglobulin H chain of catfish such as Clarias gariepinus and Clarias batrachus, but not with their light chain indicating epitope sharing among IgM H chains in these catfish. The produced polyclonal antisera were used to develop an enzyme linked immuosorbent assay to quantify IgM levels in pangasius. In conclusion, the present study provides its future implications for epidemiology and immunology studies in pangasius. 相似文献
10.
Enhancement and confirmation of white spot syndrome virus detection using monoclonal antibody specific to VP26 下载免费PDF全文
下载免费PDF全文 Akapon Vaniksampanna Siwaporn Longyant Pradit Wangman Paisarn Sithigorngul Parin Chaivisuthangkura 《Aquaculture Research》2017,48(4):1699-1710
A portion of the VP26 gene (VP26F109) encoding a structural protein of white spot syndrome virus was expressed, purified by SDS‐PAGE and used for immunization of Swiss mice for monoclonal antibody (MAb) production. Three groups of MAbs specific to different epitopes on VP26 were selected; these MAbs can be used to detect natural WSSV infection in Penaeus vannamei using dot blotting, Western blotting or immunohistochemistry without cross‐reaction with other shrimp tissues or other common shrimp viruses. The detection sensitivity of the MAbs was ranged 7–14 fmole per spot of the rVP26F109 as determined using dot blotting. A combination of three MAbs specific to VP26 with MAbs specific to VP28, VP19 and ICP11 increased the detection sensitivity of WSSV during early infection. Therefore, the MAbs specific to VP26 could be used to confirm and to enhance the detection sensitivity for WSSV infection in shrimp with various types of antibody‐based assays. 相似文献
11.
The VP 28 gene encoding a structural envelope protein of the white spot syndrome virus (WSSV) was cloned into a pET32a(+) expression vector for the production of the recombinant VP28 protein. A purified recombinant protein of 39.9 kDa size was used for polyclonal antibody production in rabbit. Specific immunoreactivity of the rabbit anti rVP28 antiserum to the viral antigen was confirmed by a Western blot. The specificity of this polyclonal anti‐rVP28 antiserum to detect the presence of the virus in WSSV‐infected Penaeus monodon was verified using a immunodot blot assay. Immunodot blot showed a positive reaction in infected shrimp tissues with prominent colour development using 3,3′,5,5′‐tetramethylbenzidine (TMB) as a chromogenic substrate when compared with 3–3′ diaminobenzidine tetrahydrochloride (DAB). Highest signal intensities of the immunodots were observed in infected shrimp pleopod extracts and haemolymph. On comparison with polymerase chain reaction (PCR), immunodot blot could detect 76% of PCR‐positive WSSV‐infected shrimp samples. Immunodot blot was found to be equivalent to first‐step PCR sensitivity to detect WSSV particles estimated to contain 1.0 × 105 viral DNA copies. 相似文献
12.
十二烷基苯磺酸钠对奥尼罗非鱼免疫毒性的研究 总被引：2，自引：0，他引：2
用5种浓度的十二烷基苯磺酸钠（sodium dodeeyl benzene sulfonic，SDBS）浸泡刺激奥尼罗非鱼（Tilapianilotica♀×raurea♂）8周，每2周采血样1次，测定免疫指标。0．05和0．10mg·L^-1组对奥尼罗非鱼的氯化硝基四氮唑兰（nitro blue tetrazolium，NBT）阳性细胞和IgM含量无影响。0．40mg·L^-1组在第8周时，IgM含量显著下降，而NBT阳性细胞突然急剧下降，与对照组相比差异极显著。0．70mg·L^-1组从第4周起，NBT阳性细胞和IgM含量开始减少，第6周时差异显著。1．00mg·L^-1组NBT阳性细胞数在第2周时量就已经有减少，但与对照组相比差异不显著，到第4周时差异极其显著；在第4周时IgM含量有显著差异，第6周后差异极显著。攻毒试验表明，0．40、0．70和1．00mg·L^-1组的死亡率均高于对照组。以上结果表明，当SDBS浓度大于0．40mg·L^-1时，奥尼罗非鱼免疫功能不同程度受到抑制，对NBT阳性细胞和IgM的影响存在着时间与剂量效应。 相似文献
13.
Chuanzhong Zhu Lijuan Yu Wei Liu Ming Jiang Shan He Ganfeng Yi Hua Wen Xufang Liang 《Aquaculture Nutrition》2019,25(6):1241-1249
A feeding trial was conducted to investigate the effect of different levels of Bacillus subtilis LT3‐1 in diets on growth, immune parameters, intestinal morphology and disease resistance in genetically improved farmed tilapia, Oreochromis niloticus. Fish (46.91 ± 0.17 g) were fed with a basal diet supplemented with B. subtilis LT3‐1 at 0 (B0), 3.8 × 1010 (B1), 7.6 × 1010 (B2), 1.14 × 1011 (B3) and 1.52 × 1011 (B4) CFU kg?1 for 6 weeks. The results showed that the weight gain of fish in B1 group was significantly enhanced compared to that in B0 group (p < 0.05). The addition of B. subtilis significantly affected serum biochemical indices (total protein, albumin, aspartate aminotransferase, alkaline phosphatase). Besides, the haematocrit, total counts of red and white blood cells, as well as the serum catalase and lysozyme activities, were increased, whereas the serum malondialdehyde, the serum immunoglobulin M and complement three contents were reduced. Parameters for intestinal morphology suggested a healthier intestine for the fish fed B. subtilis‐supplemented diets than fish fed the control diet. The survival rate after Streptococcus agalactiae challenge increased in tilapia fed with B. subtilis. The present study demonstrated B. subtilis can effectively improve growth, immunological status and resistance against S. agalactiae infection in tilapia farming. 相似文献
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分离纯化了花鲈(Lateolabrax japonicus)血清免疫球蛋白(Ig),并对其特性进行初步研究与分析.采用山羊-IgG(Goat-IgG)免疫花鲈并制备血清,分别采用硫酸铵分级沉淀法、A蛋白亲和层析法及Goat-IgG偶联亲和层析法提取花鲈Ig,对提取组份进行了蛋白浓度测定、间接ELISA效价测定、SDS-PAGE和Western-blotting分析.结果表明,采用硫酸铵分级沉淀法提取的花鲈Ig主要集中在硫酸铵饱和度为30%～50%的区间,其中45%饱和度硫酸铵富集的的抗体比活最高,ELISA效价为2×104/mg蛋白,比血清ELISA效价提高2.2倍;A蛋白柱亲和层析可得单一蛋白峰,洗脱峰集中在洗脱体积0.8～1.6 mL,处,占洗脱蛋白总量的76.2%,峰值的ELISA效价为1.37×105/mg蛋白,抗体得率为2.57%;Goat-IgG偶联Sephrose 4B亲和柱层析也可得单一蛋白峰,洗脱峰集中在洗脱体积1.4～2.8 mL处,占洗脱蛋白总量的72.4%,峰值的ELISA效价为1.39×105/mg蛋白,抗体得率为2.63%.以上3种提取方法均可得到79 kD的重链和29 kD轻链,表明可得到纯化的免疫球蛋白;采用鼠抗花鲈Ig血清和单抗AA5经Western-blotting证实提取成份为花鲈Ig.本研究表明,A蛋白柱和Goat-IgG偶联柱亲和层析均可用于花鲈Ig的提取,其中A蛋白柱法获得的洗脱峰值更为集中,抗体效价高,且提取过程不需要Goat-IgG免疫鱼体,是一种更为有效的免疫球蛋白提取方法. 相似文献
15.
温度对牙鲆皮肤黏液抗体产生的影响 总被引：1，自引：0，他引：1
在9℃、15℃、21℃和26℃4种不同水温下,用淋巴囊肿病毒(LCDV)灭活疫苗腹腔注射牙鲆,应用间接酶联免疫吸附试验(ELISA)研究了其皮肤黏液中特异性抗体水平的变化,以分析温度对牙鲆皮肤黏液抗体产生的影响.ELISA结果表明,9℃和15℃水温下牙鲆黏液OD值分别在注射LCDV后第9周和第7周达到峰值(9℃:OD=0.179; 15℃:OD=0.233); 21℃水温下OD值上升最快,5周达到峰值(0.316); 26℃水温下OD值较21℃无显著差异,也于5周达到峰值(0.295).采用硫酸铵分步盐析等技术粗提不同温度下OD值最高时的牙鲆皮肤黏液中的免疫球蛋白(Ig),SDS-PAGE检测发现,各温度组黏液蛋白中均含有72 kD和26 kD蛋白条带.Western blotting结果显示,抗牙鲆血清Ig重链的单克隆抗体只与黏液蛋白中72 kD条带发生反应,确定为牙鲆皮肤黏液Ig重链.综上结果表明,牙鲆在最适生活温度(21℃)下,抗体应答强度最大.牙鲆粗提黏液蛋白中Ig的初步确定,为探索牙鲆黏液免疫机制提供了材料. 相似文献
16.
Effect of Bacillus cereus as a water or feed additive on the gut microbiota and immunological parameters of Nile tilapia 下载免费PDF全文
下载免费PDF全文 Miao Wang Guanbin Liu Maixin Lu Xiaoli Ke Zhigang Liu Fengying Gao Jianmeng Cao Huaping Zhu Mengmeng Yi Deguang Yu 《Aquaculture Research》2017,48(6):3163-3173
We evaluated the effects of Bacillus cereus, as an additive in water and feed, on the gut microbiota and immunological parameters of Nile tilapia (Oreochromis niloticus) fingerlings. Experiments were performed in tanks and net cages respectively. Experiment 1: Tilapia were housed in tanks for 42 days, and B. cereus was added to the water at 1.0 × 104 cfu mL?1 (Treatment 1) and 1.0 ×105 cfu mL?1 (Treatment 2) weekly. For the control, no probiotic was added. Experiment 2: Tilapia were housed in cages for 42 days, and the feed was supplemented with B. cereus at 1.0 × 107 cfu g?1 (Treatment 1) and 1.0 × 108 cfu g?1 (Treatment 2) weekly. For the control, no probiotic was added. Each treatment contained three replicates, with 50 male tilapias per replicate. The fish from the probiotic treatments in both tank and cage experiments had significantly higher serum lysozyme and peroxidase activities than the control. In the cage experiment, alkaline phosphatase and total superoxide dismutase activities in tilapia were significantly higher in probiotic treatments compared with the control. The results of polymerase chain reaction‐denaturing gradient gel electrophoresis showed that B. cereus supplementation in the feed and water affected the autochthonous gut bacteria community of tilapia and stimulated various potentially beneficial bacteria. Therefore, B. cereus, as a water or feed additive, could enhance the immune status and affect the gut microbiota of tilapia. Bacillus cereus was more effective as a feed supplement rather than a water additive for enhancing the immune status of tilapia. 相似文献
17.
M A Farook N Madan G Taju S Abdul Majeed K S N Nambi N Sundar Raj S Vimal A S Sahul Hameed 《Journal of fish diseases》2014,37(8):703-710
White tail disease (WTD) caused by Macrobrachium rosenbergii nodavirus (MrNV) and extra small virus (XSV) is a serious problem in prawn hatcheries. The gene for capsid protein of MrNV (MCP43) was cloned into pRSET B expression vector. The MCP43 protein was expressed as a protein with a 6‐histidine tag in Escherichia coli GJ1158 with NaCl induction. This recombinant protein, which was used to raise the antiserum in rabbits, recognized capsid protein in different WTD‐infected post‐larvae and adult prawn. Various immunological methods such as Western blot, dot blot and ELISA techniques were employed to detect MrNV in infected samples using the antiserum raised against recombinant MCP43 of MrNV. The dot blot assay using anti‐rMCP43 was found to be capable of detecting MrNV in WTD‐infected post‐larvae as early as at 24 h post‐infection. The antiserum raised against r‐MCP43 could detect the MrNV in the infected samples at the level of 100 pg of total protein. The capsid protein of MrNV estimated by ELISA using anti‐rMCP43 and pure r‐MCP43 as a standard was found to increase gradually during the course of infection from 24 h p.i. to moribund stage. The results of immunological diagnostic methods employed in this study were compared with that of RT‐PCR to test the efficiency of antiserum raised against r‐MCP43 for the detection of MrNV. The Western blot, dot blot and ELISA detected all MrNV‐positive coded samples as detected by RT‐PCR. 相似文献
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The expression of immune‐related genes and immune responses to Aeromonas hydrophila were investigated on Oreochromis niloticus (6.07 ± 0.07 g), by feeding them six different diets for 8 weeks to apparent satiation. Diets contained fish oil (60g/kg FO), virgin coconut oil (60g/kg VCO) and corn oil (60g/kg CO) as sole lipids or blends of 30g/kg FO + 30g/kg VCO (3FVCO), 30g/kg FO + 30g/kg CO (3FCO) and 30g/kg VCO + 30g/kg CO (VO). Fish fed 3FCO recorded higher final weight, percentage weight gain (%WG) and specific growth rate (%SGR) but not significantly higher than all other groups. Triglyceride was higher in fish fed 3FCO than 3FVCO and CO (p ? 0.05), whereas total immunoglobulin (TIg) was not significant (p ? 0.05) between groups. Lysozyme activity was significantly higher in fish fed diet CO while groups FO, 3FCO and VCO recorded the least activities (p ? 0.05). Although alternative complement activity (ACH50), complement proteins (C3 and C4), was not influenced, antibody titre production was significantly higher in fish fed diet 3FCO and lower in group CO. mRNA expression of IL‐1β was significantly upregulated in fish fed VO while the expression of C‐type lysozyme and TGF‐β was not significantly influenced across treatments, although group fed FO recorded higher expression levels, respectively. Lower mortalities of fish were recorded in groups fed 3FCO and VO after 14 days postchallenge with A. hydrophila disease indicating the enhancing effects of vegetable oils to boost immune response and resistance to disease. The study concludes that alternative lipid sources with high polyunsaturated fatty acids (PUFAs‐ALA and LA) (CO) and blend of saturated fatty acids (SFA)(VCO) can partially and or exclusively replace FO with an improved effect on tilapia and resistance to A. hydrophila in tilapia. 相似文献
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鳜皮肤黏液IgM样蛋白的纯化 总被引：3，自引：0，他引：3
对经嗜水气单胞菌全菌疫苗浸泡免疫后的鳜皮肤黏液中的免疫球蛋白采用盐析法、重组蛋白A(HiTrap rProteinA Sepharose)亲和层析法及鼠抗鳜血清IgM单克隆抗体偶联的Sepharose 4B亲和层析法分离纯化,并通过SDS-PAGE及Western-blot技术对纯化蛋白的部分特性进行分析比较。结果表明:50%硫酸铵溶液可以沉淀黏液中大部分蛋白,条带仍较多,约十几条,其中含有72 ku和29 ku的条带,因此仅可作为免疫球蛋白粗提的方法;Sepharose 4B亲和层析法所提取鳜黏液Ig经SDS-PAGE检测,只含有72 ku和29 ku 2个条带(初步认为鳜黏液Ig的重链和轻链),与鳜血清IgM的重、轻链分子量相同;rProteinA亲和层析法所提蛋白除具有上述重链(72 ku)和轻链(29 ku)外,还含有43 ku的蛋白带(可能为鳜黏液Ig另外一种形式的重链)。Western-blot显示,兔抗鳜Ig多克隆抗体可与72 ku及43 ku条带发生发应。两种亲和法所提蛋白纯度较高,但含量较低,条带较淡,仅可作为实验室小量提纯鳜黏液Ig的有效方法。 相似文献