抗体

We optimized reaction system for labelling-antibody in which the optimal amount of the purified anti-Stx2B IgG conjugated with colloidal gold beads was 60μg/mL, the purified antibody for E.coli O157 was 57μg/mL, the optimal pH was 8.2, the size of colloidal gold partical was 20nm, the optimization of stabilizing agent was BSA, the optimization buffer was boracic acid buffer with pH 8.2, the optimization of preserving fluid and eluant was boracic acid buffer with pH 8.2 including 5mM NaCl-1%BSA, confining liquid for NC membrane was 0.01% PBST with 3%BSA, the amount of polyclonal antibody against E.coli O157 and Stx2 conjugated with colloidal gold beads for conjugate pad was 3μg respectively, the amount of anti-Stx monoclonal antibody for test line was 0.1μg and 1μg for E.coli O157, the amount of goat anti rabbit IgG for control line of both GICA were 1μg.

测定了胶体金颗粒的最优化反应体系：胶体金颗粒的大小为20nm；抗体与胶体金溶液结合的最佳pH约为8.2；最佳蛋白结合量分别为抗大肠杆菌O157 IgG为57μg/mL，抗重组Stx2B IgG为60μg/mL；最佳稳定剂为BSA；最佳缓冲液为pH8.2硼酸溶液；最佳金标保存液和洗涤液为5mM NaCl-1%BSA的pH 8.2的硼酸缓冲液；NC膜的封闭液为3%BSA的0.01mol/L PBST；Stx2试纸条和大肠杆菌O157试纸条的质控线上的羊抗兔IgG的多克隆抗体最佳点样量均为1μg，其检测线的抗Stx2的单克隆抗体的点样量为0.1μg、抗大肠杆菌O157的单克隆抗体的点样量为1μg，结合垫上的金标抗大肠杆菌O157和重组Stx2B多克隆抗体点样量均为3μg。

The results showed that typical FMD clinical signs produced in cattle inoculated intradermally with pathogenic FMDV after 48 h. The 3ABC antibody produced in these cattle were detected to be suspicious on the 3rd day, 3ABC antibody in the 1/4 cattle were positive on the 5th day, and the 4/4 were positive on the 8th day post virus inoculation; In 10 co-housed cattle (breeded with artificially infected pigs and sheep separately), no 3ABC antibody were detected on the 7th day and percent of positive 3ABC antibody was only 30%(3/10) on the 15th dpi (4/10 had clinical symptom).

实验表明，4头牛在舌面接种感染FMDV 48h后全数发病，第3天即出现可疑的3ABC抗体，第5天检测到阳性的3ABC抗体，至第8天3ABC抗体全部为阳性；10头同居牛（分别与接种感染猪和羊同居）在同居第7天时未检出3ABC抗体，在同居第15天时，3ABC抗体阳性率仅为30%；肌肉接种病毒羊3ABC抗体产生时间与接种病毒牛一致。

Citri had been selected by ribosome display in previous research. In order to express the scFvs with bio-activity efficiently, the genes of the selected scFvs were transformed into Escherichia coli HB2151 for expressing resoluble antibodies. The expression levels of scFvs were detected by dot blotting and Western blotting, and the expressed scFvs were purified by affinity chromatography, and then the activity of the purified scFvs was tested by enzyme-linked immunosorbent assay .

Citri， Xac高亲和力单链抗体(Single chain variable fragment， scFv)GX13、GX44和GX95，为高效表达具有生物活性的单链抗体，本研究将前期筛选的高亲和力单链抗体基因转入大肠杆菌HB2151中进行可溶性表达，点印迹和Western印迹检测可溶性单链抗体的表达水平，亲和层析纯化表达的单链抗体，酶联免疫吸附法检测纯化单链抗体的抗原结合活性。

ELISA showed that antibody responses induced by propolis vaccine were faster than that of oil-emulsion vaccines and aluminum hydroxide vaccines;antibody responses induced by oil-emulsion vaccines were longer in duration than that of propolis vaccines and aluminum hydroxide vaccines;the maximum antibody responses induced by oil-emulsion vaccines and propolis vaccines were higher than that of aluminum hydroxide vaccines;Antibody levels were all significantly boosted by revaccination of these vaccines and in addition to this,oil-emulsion vaccines showed longer antibody duration.

这七种疫苗在抗体产生速度方面以蜂胶苗较快，而油剂苗和铝胶苗免疫后抗体上升速度较慢；在抗体水平的维持能力方面，油剂苗较蜂胶苗和铝胶苗更强；在产生最高抗体水平方面，油剂苗和蜂胶苗能力相当且较铝胶苗为高；在加强免疫的效果方面，这些疫苗二次免疫所产生的抗体水平均较一次免疫更高，蜂胶苗和铝胶苗在第93天时的二免抗体水平已降至同一免相当，但此时油剂苗有着较一免更高的二免抗体水平。

Of the patients tested using CDC technology, 17 (3.8%) had a panel reactive antibody higher than 10% and 8 patients (1.83%) had a PRA higher than 25% for class I antibodies. Class I sensitized patients, defined as patients with a PRA higher than 10%, had significantly lower survival rates than class I nonsensitized patients ( P =.0035). Class I nonsensitized patients had survival rates of 79.4% at 1 year posttransplant, 63.2% at 3 years, and 45.4% at 5 years. Sensitized patients had 1, 3, and 5 year survival rates of 64.7%, 28.8%, and 9.6%, respectively.

使用CDC检测的病患之中，有17位(3.8%)群体反应抗体第一类抗体高於10%、有8位(1.83%) PRA超过25%，第一类抗体敏感病患定义为PRA高於10%者，相较於第一类抗体不敏感病患、存活率显著降低(P =。0035)；第一类抗体不敏感病患移植后一年之存活率为79.4%、3年存活率为63.2%、5年存活率为45.4%；第一类抗体敏感病患移植后一年、3年、5年之存活率分别为64.7%、28.8%、9.6%。

The blots were probed with anti-Cdx-2, anti-JNK (Santa Cruz Biotechnology, Santa Cruz, California), anti-ERK, anti-phospho -ERK, antip38MAPK,anti-p-p38MAPK, anti-p-JNK (Cell Signaling, Beverly, Massachusetts) and anti-actin (Amersham Life Sciences,Piscataway, New Jersey) antibodies, and then incubated with appropriate species specific secondary antibodies labeled with horseradish peroxidase.

Blots分别与抗CDX-2抗体，抗JNK抗体（圣克鲁斯生物技术，圣克鲁斯，加州），抗ERK抗体，抗phospho -ERK抗体，抗p38MAPK抗体，抗p-p38MAPK抗体，抗p-JNK抗体（Cell Signaling， Beverly，美国马萨诸塞州）和抗actin抗体（ Amersham生命科学，皮斯卡塔韦，新泽西州）作用，然后在与辣根过氧化物酶标记的第二抗体孵育。

In addition, the term antibody as used herein also encompasses various types of antibodies, including human, chimeric, humanized, primatized, veneered or single chain antibodies.

而且，本文所用的术语抗体也包括多种抗体，其包括人抗体、嵌合抗体、人化抗体、灵掌源抗体、veneered 或单链抗体。

objective to establish immunological methods specific for detecting antigens in different groups of monoclonal antibodies.methods indirect immnofluorescence assay was applied to identify specificity of the two groups of monoclonal antibodies prepared with crude antigen and recombinant antigen of aspergillus fumigatus,respectively.two different double monoclonal antibody sandwich elisa assays established with the two groups of antibodies were performed to detect antigents in the cell culture supermatants of 19 common species of aspergillus,penicillium marneffei,and 5 species of candidas.results the results of indirect immnofluorescence assay indicated that the monoclonal antibodies prepared with crude antigen of aspergillus fumigatus were specific for antigens in both clinical isolates and environmental isolates of aspergillus, whereas the other group of monoclonal antibodies was proved to be specific for aspergillus fumigatus of both clinical and environmental isolates.the elisa assay established with the crude antigen-specfic monoclonal antibodies could detect both of the clinical and environmental isolates of aspergllius, while the other assay could only detect aspergillus fumigatus of both clinical and environmental isolates.and no cross reaction with the cell culture of penialllium marneffei and candidas was observed with the two methods.conclusion the elisa assays can detect both of the clinical and environmental isolates of aspergillus,and differentiate aspergillus fumigatus from other species of aspergillus.

目的 用2组曲霉单克隆抗体建立特异性识别不同种类曲霉抗原的检测方法。方法采用天然烟曲霉抗原免疫，获得广谱针对曲霉抗原的单克隆抗体；采用重组烟曲霉抗原获得特异性针对烟曲霉抗原的单克隆抗体，用间接免疫荧光鉴定，并分别建立2种双抗体夹心elisa法，对19种常见的环境和临床分离曲霉株、马尔尼菲氏青霉菌及念珠菌培养液进行检测。结果间接免疫荧光显示，用天然烟曲霉抗原免疫获得的单克隆抗体(mabs-1)可广谱识别多种曲霉分离株，而重组烟曲霉抗原获得的单克降抗体(mabs-2)仅能特异性结合临床和环境分离的烟曲霉抗原。用mabs-1建立的双抗体夹心elisa法可检测19种常见曲霉株培养液；用特异性针对烟曲霉抗原单克降抗体(mabs-2)建立的双抗体夹心elisa法可特异性检测临床和环境分离株烟曲霉培养液；与其他曲霉株无交叉反应；2种双抗体夹心elisa法与马尔尼菲氏青霉菌及念珠菌培养液均无交叉反应。

Pneumophila serogroup 5 antibody to develop two immuo-colloidal gold tests respectively. Main works listed below as:1、Preparation and identification of polysaccharide antigen of L. pneumophila Bacteria of LP1 ~ 7,9 and 10 were cultured on yeast agar buffer activated carbon for three to five days at 37℃, 5%CO2. Protein-free polysaccharide antigens were obtained after harvest in cells, extraction, deproteinization, dialysis and other steps. Their immunogenicities were verified by ultraviolet spectrophotometer full-wavelength scanning and Western blotting.2、Preparation and identification of rabbit anti-LP1 antibodies and rabbit anti-LP5 antibody Rabbit anti-LP1 and anti-LP5 antibodies were purified after rabbits were immuned with antigens isolated as described above. The purities of both antibodies were above 80% and the titer of blood serum 1:32 tested by double antibody sandwich assay.3、Development of colloidal gold immunochromatographic assay kit The size of colloidal gold particles in the kit was 25nm. The optimal concentrations for antibodies were 30μg/ml and the sensitized concentrations of NC membrane were 5 mg/ml.

主要研究工作从以下几个方面进行：1、LP1~7、9和10型多糖抗原的制备与鉴定将LP1~7、9和10型菌株分别接种在缓冲活性炭酵母琼脂培养基上，37℃、5%CO2的条件下培养，3~5天后洗下菌苔，经抽提、除蛋白、透析等步骤后得到基本无蛋白的LP多糖抗原，经紫外分光光度仪全波长扫描及Western blotting验证其抗原良好。2、兔抗LP1抗体和兔抗LP5抗体的制备与鉴定分别用LP1、LP5型多糖抗原免疫家兔，采用琼脂糖双向扩散试验检测，两种抗体血清效价均为1：32；饱和硫酸铵法提取抗体，SDS-PAGE检定其抗体纯度均达到80%以上。3、胶体金免疫层析检测试剂的初步研制采用柠檬酸三钠还原法制备约25nm大小的胶体金颗粒；分别制备兔抗LP1抗体、兔抗LP5抗体的金标探针，两种抗体的最适标记量均为30μg/ml；选择适当孔径的微孔滤膜为载体包被两种抗体，NC膜包被浓度均为5mg/ml。

The invention provides a human McAb to tetanus toxin, being a human antibody, which consists of an antibody variable region and an antibody constant region or not, having the ability of neutralizing the tetanus toxin, with the variable region gene and protein sequence indicated as the sequence table 1. The invention is characterized in that the antibody can not induce obvious allergic reaction, has higher titer and longer effect without any animal virus pollution, and can be produced in an unlimited quantity, which also provides the relating biological functions, the testing methods, the manufacturing method and the application.

A 本发明提供一种人源抗破伤风毒素单克隆抗体，其为人源抗体，包括抗体可变区，该抗体包括或不包括抗体的恒定区，该抗体具有中和破伤风毒素的能力，抗体可变区的基因和蛋白质序列如序列表1所示；其具有不会诱发明显的过敏反应，有较高的效价且长效，产品无动物病毒污染，可以无限量地生产；本发明还提供了其生物功能、测定方法、生产制造方法及其应用。

This one mode pays close attention to network credence foundation of the businessman very much.

这一模式非常关注商人的网络信用基础。

Cell morphology of bacterial ghost of Pasteurella multocida was observed by scanning electron microscopy and inactivation ratio was estimated by CFU analysi.

扫描电镜观察多杀性巴氏杆菌细菌幽灵和菌落形成单位评价遗传灭活率。