期刊文献+
共找到770篇文章
< 1 2 39 >
每页显示 20 50 100
Expression and identification of recombinant soluble single-chain variable fragment of monoclonal antibody MC3 被引量:13
1
作者 Feng-Tian He Rong-Fen Li Yun-Sheng Kang Yan Zhang,Department of Biochemistry & Molecular Biology,Third Military Medical University,Chongqing 400038,China Yong-Zhan Nie Bao-Jun Chen Tai-Dong Qiao Dai-Ming Fan,Institute of Digestive Disease,Xijing Hospital,Fourth Military Medical University,Xi’an 710032,Shaanxi Province,China 《World Journal of Gastroenterology》 SCIE CAS CSCD 2002年第2期258-262,共5页
AIM: To generate soluble single chain variable fragments (ScFv) of monoclonal antibody MC3 recognizing colorectal and gastric carcinomas. METHODS: mRNA was isolated from the hybridoma cell line producing MC3 and the D... AIM: To generate soluble single chain variable fragments (ScFv) of monoclonal antibody MC3 recognizing colorectal and gastric carcinomas. METHODS: mRNA was isolated from the hybridoma cell line producing MC3 and the DNAs encoding variable domains of heavy and light chains (VH and VL) of the antibody were amplified separately by RT-PCR and assembled into ScFv DNA with a linker DNA. The ScFv DNA was ligated into the phagemid vector pCANTAB5E and the ligated sample was transformed into E.coli TG1.The transformed cells were infected with M13KO7 helper phage to yield recombinant phages. After two rounds of panning with gastric carcinoma cell line AGS highly expressing MC3-binding antigen, the phage clones displaying ScFv fragments of the antibody were selected by ELISA. 4 phage clones showing strong signal in ELISA were used to infect E.coli HB2151 to express soluble ScFvs. The soluble ScFvs were identified by Dot blot and Western blot, and their antigen-binding activity was assayed by ELISA. The VH and VL DNAs of the ScFv DNA derived from phage clone 19 were sequenced. RESULTS: The VH,VL and ScFv DNAs were about 340 bp, 320 bp and 750 bp respectively. After two rounds of panning to the recombinant phages, 18 antigen-positive phage clones were selected from 30 preselected phage clones by ELISA. All the soluble ScFvs derived from the 4 out of the 18 antigen-positive phage clones were about M(r)32000 and concentrated in periplasmatic space under the given culture condition. The soluble ScFvs could bind the antigen, and they shared the same binding site with MC3. The sequences of the VH and VL DNAs of the MC3 ScFv showed that the variable antibody genes belonged to the IgG1 subgroup,kappa-type. CONCLUSION: The soluble ScFv of MC3 is successfully produced, which not only provides a possible novel targeting vehicle for in vivo and in vitro study on associated cancers, but also offers the antibody a stable genetic source. 展开更多
关键词 Animals Antibodies Monoclonal Base Sequence Carcinoma Colorectal Neoplasms Enzyme-Linked Immunosorbent Assay Humans Immunoglobulin Fragments Immunoglobulin Variable Region Mice Molecular Sequence Data recombinant Proteins Stomach Neoplasms Tumor Cells Cultured
下载PDF
Recombinant Human IgG antibodies against Human Cytomegalovirus 被引量:1
2
作者 TAO DUAN XIAO-FANG WANG +2 位作者 SHU-YUAN XIAO SHU-YAN GU AND MI-FANG LIANG 《Biomedical and Environmental Sciences》 SCIE CAS CSCD 2008年第5期372-380,共9页
Objective To study the passive immunization with human monoclonal antibodies as for prophylaxis of human cytomegalovirus (HCMV) infection. Methods Fab monoclonal antibodies to HCMV were recovered by repertoire cloni... Objective To study the passive immunization with human monoclonal antibodies as for prophylaxis of human cytomegalovirus (HCMV) infection. Methods Fab monoclonal antibodies to HCMV were recovered by repertoire cloning of mRNA from a HCMV infected individual. Antigen binding specificity, CDR sequence of VH and VL and neutralizing activity on HCMV AD169 stain were analyzed in vitro. The light and heavy chain Fd fragment genes of Fab antibodies were further cloned into a recombinant baculovirus expression vector pAC-K-Fc to express intact IgG. Secreted products were purified with affinity chromatography using protein G. Results SDS-PAGE and Western blot confirmed the expression of the intact IgG. Immuno-blotting and -precipitation were used to identify HCMV proteins. One Fab monoclonal antibody recognized a conformational HCMV protein. Conclusion IgG antibodies can neutralize the HCMV AD169 strain efficiently at a titer of 2.5 μg/mL and may prove valuable for passive immunoprophylaxis against HCMV infection in humans. 展开更多
关键词 Human cytomegalovirus Human engineering antibody Phage display recombinant baculovirus expression
下载PDF
Neutralizing Antibody Titer Test of Ebola Recombinant Protein Vaccine and Gene Vector Vaccine pVR-GP-FC 被引量:1
3
作者 YANG Ren ZHU Ying +8 位作者 MA Jing HAO Yan Zhe WANG Xuan HOU Mei Ling LIU Li Peng FAN Li Yun CAO Yu Xi ZHANG Xiao Guang LI Xiao Jing 《Biomedical and Environmental Sciences》 SCIE CAS CSCD 2018年第10期721-728,共8页
Objective In previous studies, we immunized mice with Ebola recombinant protein vaccine and gene vector vaccine. Both stimulated high levels of humoral immunity. In this work, we constructed a pseudovirus containing E... Objective In previous studies, we immunized mice with Ebola recombinant protein vaccine and gene vector vaccine. Both stimulated high levels of humoral immunity. In this work, we constructed a pseudovirus containing Ebola membrane proteins to verify whether the two immunization strategies can induce neutralizing antibodies in mice. Methods A pseudovirus containing an Ebola virus membrane protein based on the HIV-1 viral gene sequence was constructed and evaluated using a known neutralizing antibody. The titer of the neutralizing antibody in the sera of mice immunized with the recombinant protein and the gene vector vaccine was examined using a neutralization test. Results Ebola pseudovirus was successfully prepared and applied for neutralizing antibody detection. Immunological experiments showed that recombinant protein GP-Fc and gene vaccine pVR-modGP-Fc had good immunogenicity. The titer of the bound antibody in the serum after 8 weeks of immunization in mice was more than 1:105, and the recombinant protein induced greater humoral immunity. The results of the neutralization test based on the Ebola pseudovirus system demonstrated that both vaccines induced production of protective antibodies, while the gene vaccine induced a higher titer of neutralizing antibodies. Conclusion An Ebola pseudovirus detection system was successfully established and used to evaluate two Ebola vaccines. Both produced good immunogenicity. The findings lay the foundation for the development of new Ebola vaccines and screening for neutralizing monoclonal antibodies. 展开更多
关键词 Ebola virus recombinant subunit vaccine DNA vaccine Neutralizing antibody
下载PDF
Rabies Virus Neutralizing Activity,Safety,and Immunogenicity of Recombinant Human Rabies Antibody Compared with Human Rabies Immunoglobulin in Healthy Adults 被引量:2
4
作者 ZHANG Jun Nan MENG Ya Juan +16 位作者 BAI Yun Hua LI Yu Feng YANG Li Qing SHI Nian Min HAN Hui Xia GAO Jian ZHU Li Juan LI Shu Ping ZHANG Jing ZHAO Qin Hua WANG Xiu Qin WEI Jing Shuang REN Le Min CAO Chen Hua CHEN Chen ZHAO Wei LI Li 《Biomedical and Environmental Sciences》 SCIE CAS CSCD 2022年第9期782-791,共10页
Objective Preliminary assessment of rabies virus neutralizing activity,safety and immunogenicity of a recombinant human rabies antibody(NM57)compared with human rabies immunoglobulin(HRIG)in Chinese healthy adults.Met... Objective Preliminary assessment of rabies virus neutralizing activity,safety and immunogenicity of a recombinant human rabies antibody(NM57)compared with human rabies immunoglobulin(HRIG)in Chinese healthy adults.Methods Subjects were randomly(1:1:1)allocated to Groups A(20 IU/kg NM57),B(40 IU/kg NM57),or C(20 IU/kg HRIG).One injection was given on the day of enrollment.Blood samples were collected on days-7 to 0(pre-injection),3,7,14,28,and 42.Adverse events(AEs)and serious AEs(SAEs)were recorded over a period of 42 days after injection.Results All 60 subjects developed detectable rabies virus neutralizing antibodies(RVNAs)(>0.05 IU/mL)on days 3,7,14,28,and 42.The RVNA levels peaked on day 3 in all three groups,with a geometric mean concentration(GMC)of 0.2139 IU/mL in Group A,0.3660 IU/mL in Group B,and0.1994 IU/mL in Group C.At each follow-up point,the GMC in Group B was significantly higher than that in Groups A and C.The areas under the antibody concentration curve over 0-14 days and 0-42 days in Group B were significantly larger than those in Groups A and C.Fifteen AEs were reported.Except for one grade 2 myalgia in Group C,the other 14 were all grade 1.No SAEs were observed.Conclusion The rabies virus neutralizing activity of 40 IU/kg NM57 was superior to that of 20 IU/kg NM57 and 20 IU/kg HRIG,and the rabies virus neutralizing activity of 20 IU/kg NM57 and 20 IU/kg HRIG were similar.Safety was comparable between NM57 and HRIG. 展开更多
关键词 recombinant human rabies antibody NM57 Human rabies immunoglobulin Rabies virus neutralizing activity SAFETY IMMUNOGENICITY
下载PDF
Isolation and Characterization of Recombinant Variable Domain of Heavy Chain Anti-idiotypic Antibodies Specific to Aflatoxin B_1 被引量:2
5
作者 WANG Dan XU Yang +5 位作者 TU Zhui FU Jin Heng XIONG Yong Hua FENG Fan TAO Yong LEI Da 《Biomedical and Environmental Sciences》 SCIE CAS CSCD 2014年第2期118-121,共4页
Some unique subclasses of Camelidae antibodies are devoid of the light chain, and the antigen binding site is comprised exclusively of the variable domain of the heavy chain (VHH). The recombinant VHHs have a high p... Some unique subclasses of Camelidae antibodies are devoid of the light chain, and the antigen binding site is comprised exclusively of the variable domain of the heavy chain (VHH). The recombinant VHHs have a high potential as alternative reagents for the next generation of immunoassay. In particular, they might be very useful for molecular mimicry. The present study demonstrated an alpaca immunized with the F(ab')z fragment of anti-aflatoxin B1 mAb and developed an important anti-idiotypic (anti-ld) responses. Antigen-specific elution method was used for panning private anti-ld VHHs from the constructed alpaca VHH library. The selected VHHs were expressed, renatured, purified, and then identified by a competitive enzyme-linked immunosorbent assay (ELISA). Our findings indicated that the VHH would be an alternative tool for haptens mimicry studies. 展开更多
关键词 ab VHH Isolation and Characterization of recombinant Variable Domain of Heavy Chain Anti-idiotypic Antibodies Specific to Aflatoxin B1
下载PDF
PURIFICATION OF RECOMBINANT HUMAN INTERFERON-γ BY IMMUNOAFFINITY CHROMATOGRAPHY WITH MONOCLONAL ANTIBODY
6
作者 丛进阳 陈薇 《Chinese Journal of Chemical Engineering》 SCIE EI CAS CSCD 1995年第3期4-12,共9页
E.coli cells expressing recombinant human interferon-γ was disrupted by sonication anddissolved in 7mol·L<sup>-1</sup> guanidine hydrochloride.The extract obtained was then renaturated by 70 folddilu... E.coli cells expressing recombinant human interferon-γ was disrupted by sonication anddissolved in 7mol·L<sup>-1</sup> guanidine hydrochloride.The extract obtained was then renaturated by 70 folddilution with PBS.HulFN γ was purified by affinity chromatography with monoclonal antibody fromthe renaturated crude feed solution.After washing the column with PBS,the adsorbed HulFN γ waseluted with PBS containing 0.5mol·L<sup>-1</sup> NaCl.The column was regenerated with 2mol·L<sup>-1</sup> GuHClfor reuse.After one step of affinity purification the purity of interferon-γ was over 95%.and thespecific activity of the HulFN-γ reached 1.2×10<sup>7</sup> IU·mg<sup>-1</sup> protein.92.8% of recovery was obtainedin the elution step.Total recovery of HulFN γ activity in the affinity chromatography was 78%. 展开更多
关键词 MONOCLONAL antibody AFFINITY chromatographv recombinant human INTERFERON
下载PDF
Production and Characterization of Monoclonal Antibody Against Recombinant Human Erythropoietin
7
作者 JIE-BO MI JIN YAN +3 位作者 XIAO-JIE DING ZHEN-QUAN GUO MEI-PING ZHAO WEN-BAO CHANG 《Biomedical and Environmental Sciences》 SCIE CAS CSCD 2007年第3期184-188,共5页
Objective To produce specific monoclonal antibody (mAb) against recombinant human erythropoietin (rHuEPO) for development of highly efficient methods for erythropoietin detection in biological fluids. Methods rHuE... Objective To produce specific monoclonal antibody (mAb) against recombinant human erythropoietin (rHuEPO) for development of highly efficient methods for erythropoietin detection in biological fluids. Methods rHuEPO was covalently coupled with bovine serum albumin (BSA) and the conjugate was used to immunize mice to produce specific mAb against rHuEPO based on hybridoma technology. The obtained F3-mAb was characterized by enzyme-linked irmnunosorbent assay (ELISA), SDS-PAGE and Western blot. Results The isotype of F3-mAb was found to be IgM with an affinity constant of 2.1x10s L/mol. The competitive ELISA using the obtained IgM showed a broader linear range and lower detection limit compared with previous work. Conclusions The modification of rHuEPO was proved to be successful in generating required specific mAb with high avidity to rHuEPO. 展开更多
关键词 recombinant human erythropoietin Monoclonal antibody IGM ELISA
下载PDF
Immune Blot Analysis on Expression of the Mammalian Target of Rapamycin in Goat Fetal Fibroblasts with Recombinant Polyclonal Antibody
8
作者 LIANG Yan LI Shu-yu +8 位作者 WANG Xiao-jing WU Man-lin YANG Jiao-fu LI Jie HAO Xi-yan HAO Hui-fang BA Yin-ma CHEN Xian-wei WANG Zhi-gang 《Journal of Integrative Agriculture》 SCIE CSCD 2012年第6期1002-1008,共7页
To detect the mammalian target of rapamycin (mTOR) expressed in Cashmere goat fetal fibroblasts (GFb), mTOR gene was cloned from Inner Mongolia Cashmere goat (Capra hircus) and expressed in Escherichia coli foll... To detect the mammalian target of rapamycin (mTOR) expressed in Cashmere goat fetal fibroblasts (GFb), mTOR gene was cloned from Inner Mongolia Cashmere goat (Capra hircus) and expressed in Escherichia coli followed by immunizing mice with the purified recombinant protein as an irnmunogen to produce the anti-goat mTOR recombinant polyclonal antibody. Antiserum was collected from the immunized mice after the fifth immunization and its titer was determined with enzyme-linked immunosorbent assay (ELISA). The results showed that the recombinant polyclonal antibody had a titer 1:200000 and could react with the roTOR expressed in GFb cells with a specific and sensitive affinity. Western blot showed that mTOR expression and phospho-mTOR (Ser 2448) activity were inhibited when GFb cells were treated with CCI-779, an mTOR specific inhibitor. 展开更多
关键词 prokaryotic expression recombinant antibody GOAT mTOR CCI-779
下载PDF
Detection of Antibodies against Toxoplasma gondii by ELISA with Recombinant Microneme Protein 3
9
作者 JIANG Tao YAO Bao-an ZHAO Jun-long 《Animal Husbandry and Feed Science》 CAS 2009年第8期30-31,39,共3页
[Objective] To develop a new method for serodiagnosis of swine toxoplasmosis. [Method] With the purified recombinant microneme protein 3 (rMIC3) as coating antigens, an indirect ELISA was developed for detection of ... [Objective] To develop a new method for serodiagnosis of swine toxoplasmosis. [Method] With the purified recombinant microneme protein 3 (rMIC3) as coating antigens, an indirect ELISA was developed for detection of antibodies against Toxoplasma gondii. [ Result] The optimal working concentration of rMIC3 was 3. 40 ug/ml, and the optimal degree of dilution of sera was 1:160. Cross-reaction was not observed between the Toxoplasma gondii-positive sera and the positive sera against classical swine fever virus or some other pathogens. The developed ELISA had 92.56% coincidence rate with latex agglutination test. [ Conclusion] The developed ELISA is sensitive, rapid, specific and reproducible, and thus it can be applied in serodiagnosis and seroprevalence investigation of swine toxoplasmosis. 展开更多
关键词 Toxoplasma gondii recombinant microneme protein 3 Enzyme linked immunosorbent assay ANTIBODIES
下载PDF
Anti-VDAC3 recombinant antibody decreased human sperm motility and membrane integrity: A potential spermicide for contraception
10
作者 Asmarinah Tri Panjiasih Susmiarsih +3 位作者 Amalia Shari Putri Ratri Dwi Ari Pujianto Endang Winiati Bachtiar 《Asian pacific Journal of Reproduction》 2017年第6期257-263,共7页
Objective:To express recombinant protein that comprises an important fragment of human sperm specific voltage dependent anion channel 3 (VDAC3) protein as a potential molecule for generation of antibody, which can aff... Objective:To express recombinant protein that comprises an important fragment of human sperm specific voltage dependent anion channel 3 (VDAC3) protein as a potential molecule for generation of antibody, which can affect sperm function, aiming at spermicide development. Methods: The produce of VDAC3 recombinant protein encoded by cDNA sequence of human VDAC3 exon 5-8, based on experimental design of VDAC3 knock-out mice study. And after the purification of various human sperm VDAC3 recombinant proteins, epitope has been predicted in our recombinant protein determined by ElliPro program. Polyclonal antibody was produced for 14 wk. Then anti-VDAC3-exon 5-8 recombinant antiserum was inoculated to human sperm. After the process, antibody VDAC3 protein in human sperm was incubation with anti-VDAC3 recombinant antibody. Finally evaluation the effect of VDAC3 antiserum to human sperm motility and plasma membrane integrity was proceeded.Results: Human VDAC3 recombinant protein was successfully over-expressed in Escherichia coli and purified by affinity chromatography method. Purified human sperm VDAC3 recombinant protein could stimulate immune response in rabbit producing an antibody against VDAC3. Anti-VDAC3 recombinant antibody recognized VDAC3 antigen in human sperm could decrease human sperm motility and membrane integrity significantly.Conclusions:Anti-VDAC3 recombinant polyclonal antibody that we produced in rabbit by ourselves could decrease sperm motility and sperm membrane integrity. The authors suggest this polyclonal antibody could be used as a candidate agent for male contraception in the future. Furthermore, the authors intend to explore the effect of this antibody into sperm function aiming at male contraceptive vaccine development. 展开更多
关键词 recombinant VDAC3 antibody SPERM MOTILITY Membrane integrity SPERMICIDE
下载PDF
SYNTHESIS OF INTERFERON-α_A MONOCLONAL ANTIBODY PACKING MATERIAL IN HIGH-PERFORMANCE AFFINITY CHROMATOGRAPHY AND PURIFICATION OF RECOMBINANT HUMAN INTERFERON-α_A
11
作者 Wen Ke FENG Xin Du GENG Laboratory of Modern Separation Science, Department of Chemistry Northwest University, Xi’an 710069 《Chinese Chemical Letters》 SCIE CAS CSCD 1991年第5期383-386,共4页
A new way for the synthesis of human interferon—α_A monoclonal antibody (IFN-α_A-McAb) bound to silica gel packing material in high-performance affinity chromatography (HPAFC) has been developed. The high coupling ... A new way for the synthesis of human interferon—α_A monoclonal antibody (IFN-α_A-McAb) bound to silica gel packing material in high-performance affinity chromatography (HPAFC) has been developed. The high coupling efficiency and specific activity of IFN—α_A-McAb can be obtained by activated diol-silica gel with activating agent. After purification using this packing material in HPAFC, the specific activity of recombinant human interferon-α_A (rIFN-α_A) rose up to 1.03×10~7IU/mg protein and the purification efficiency is appoximately 100 times. 展开更多
关键词 IFN SYNTHESIS OF INTERFERON A MONOCLONAL antibody PACKING MATERIAL IN HIGH-PERFORMANCE AFFINITY CHROMATOGRAPHY AND PURIFICATION OF recombinant HUMAN INTERFERON
下载PDF
Recombinant IgM expression in mammalian cells: A target protein challenging biotechnological production 被引量:3
12
作者 Alexander Mader Veronika Chromikova Renate Kunert 《Advances in Bioscience and Biotechnology》 2013年第4期38-43,共6页
For many years, the potential of immunoglobulin M (IgM) antibodies was not fully understood because of characteristics different to the well-known immunoglobulin G type like low target affinity, cross reactivity and c... For many years, the potential of immunoglobulin M (IgM) antibodies was not fully understood because of characteristics different to the well-known immunoglobulin G type like low target affinity, cross reactivity and complex protein structure. In the meanwhile IgMs have been positively evaluated for their use as therapeutic agent in the mucosal environment but also in serum to eradicate upcoming tumor cells and invading antigens. Therefore IgM class of antibodies will play a significant role in clinical applications but also diagnosis in the future. To evaluate the full potential of this kind of antibody molecules large amounts of high quality product will be needed. In this review the focus is set on the biotechnological aspect of producing IgM class antibodies recombinantly in mammalian cells. Current achievements in expression and purification of this molecule are highlighted and compared. 展开更多
关键词 IGM MAMMALIAN EXPRESSION recombinant antibody EXPRESSION antibody Purification
下载PDF
Study of Immunoassay Methods for Recombinant Human Erythropoietin (rhEPO) Using Competitive ELISA
13
作者 JinYAN JieBoMI WenBaoCHANG 《Chinese Chemical Letters》 SCIE CAS CSCD 2004年第8期939-942,共4页
Two different immunoassay methods, competitive indirect enzyme-linked immuno-sorbent assay (CI-ELISA) and amplificative competitive indirect ELISA (ACI-ELISA) using biotin-avidin complex system were studied to detect ... Two different immunoassay methods, competitive indirect enzyme-linked immuno-sorbent assay (CI-ELISA) and amplificative competitive indirect ELISA (ACI-ELISA) using biotin-avidin complex system were studied to detect rhEPO. The linear ranges were 50-20000 ng/mL and 10-50000 ng/mL for CI-ELISA and ACI-ELISA, respectively. The low detection limits of CI-ELISA and ACI-ELISA were 62.8 ng/mL and 8.5 ng/mL, respectively. 展开更多
关键词 recombinant human erythropoietin (rhEPO) polyclonal antibody (pAb) competitive indirect ELISA (CI-ELISA) ampliflcative competitive indirect ELISA (ACI-ELISA).
下载PDF
鼠疫耶尔森菌低钙应答V抗原人源单克隆抗体筛选与鉴定
14
作者 张黎 郑滨洋 +5 位作者 张琪 吴海莲 潘红星 朱凤才 吴海生 周剑芳 《中国人兽共患病学报》 CAS CSCD 北大核心 2024年第1期15-20,共6页
目的筛选抗低钙应答V抗原(LcrV)的人源单克隆抗体,分析抗体基因特点,检测抗体与抗原结合的特异性、亲和力及功能。方法使用PCR方法从鼠疫疫苗临床试验志愿者外周血细胞cDNA扩增抗体轻、重链可变区基因片段,构建ScFv噬菌体抗体文库。用L... 目的筛选抗低钙应答V抗原(LcrV)的人源单克隆抗体,分析抗体基因特点,检测抗体与抗原结合的特异性、亲和力及功能。方法使用PCR方法从鼠疫疫苗临床试验志愿者外周血细胞cDNA扩增抗体轻、重链可变区基因片段,构建ScFv噬菌体抗体文库。用LcrV对文库进行富集筛选,将获得的抗体基因表达成人IgG1后,测定抗体与LcrV抗原结合的特异性、亲和力、免疫调节功能以及保护能力。结果成功构建了鼠疫耶尔森菌人源ScFv抗体文库,库容量为7.54×10^(8)。抗体文库经LcrV筛选后获得了3株抗LcrV抗体,命名为RV-B4、RV-D1和RV-E8。3株抗体重链为VH1-46和VH3-30,轻链分别为VL1-51、VK3-20和VK1-39。3株抗体经ELISA及Western blot验证均与LcrV特异性结合,其与V抗原结合的解离常数(KD)分别为2.1 nmol/L、1.24 nmol/L和42 nmol/L。RV-D1体外降低人THP-1分泌TNF-α,动物攻毒试验中未发现抗LcrV抗体的保护效果。结论从鼠疫疫苗免疫人群获得了靶向低钙应答V抗原的人源抗体,以结合抗体为主,不能有效阻断病菌感染。所获得的单抗为鼠疫免疫基础研究、鼠疫诊断应用提供了候选材料。 展开更多
关键词 鼠疫耶尔森菌 低钙应答V抗原 噬菌体展示 重组抗体 保护效果
下载PDF
RhC抗原弱表达伴类自身抗-Ce及同种抗-Jkb致配血不合分析
15
作者 杨红梅 虞茜 +3 位作者 邹昕 马思飞 陈瑾 张建伟 《中国实验血液学杂志》 CAS CSCD 北大核心 2024年第5期1539-1544,共6页
目的:探讨1例患者类自身抗-Ce合并抗-Jkb引起交叉配血不合并分析其RHCE基因弱表达的原因。方法:采用试管法、毛细管离心法对ABO、Rh和Kidd血型抗原进行鉴定。采用盐水、聚凝胺、抗人球蛋白三介质联用多套谱细胞进行抗体筛查及抗体特异... 目的:探讨1例患者类自身抗-Ce合并抗-Jkb引起交叉配血不合并分析其RHCE基因弱表达的原因。方法:采用试管法、毛细管离心法对ABO、Rh和Kidd血型抗原进行鉴定。采用盐水、聚凝胺、抗人球蛋白三介质联用多套谱细胞进行抗体筛查及抗体特异性鉴定;采用多重PCR技术对RHCE基因进行测序及单倍体分析并使用SwissModel进行RHCE蛋白建模。结果:该患者血清中检出类自身抗-Ce合并抗-Jkb抗体。RHCE三代单分子测序显示突变组合为c.48G>C、c.150C>T、c.178C>A、c.201A>G、c.203A>G和c.307C>T,在内含子2中存在109 bp插入序列,同时出现内含子5-8大片段丢失,其Rh血型基因型为DCe/DCe,表型为CCDee。结论:基因分型技术可以协助推断患者血清中部分弱表达的RhC、c、E、e的分子机理,以辅助疑难抗体的鉴定,从而保证患者输血安全。 展开更多
关键词 基因重组 抗体筛查 类自身抗-Ce 抗-Jkb
下载PDF
重组人Ⅱ型肿瘤坏死因子受体-抗体融合蛋白治疗中毒性表皮坏死松解症的疗效及安全性
16
作者 王燕玲 王丽娜 +5 位作者 黄巧玲 王燕燕 宋娜娜 刘旭蓉 吴静 蔡兴锐 《临床和实验医学杂志》 2024年第12期1265-1268,共4页
目的探讨重组人Ⅱ型肿瘤坏死因子受体-抗体融合蛋白(rhTNFR:Fc)治疗中毒性表皮坏死松解症(TEN)患者的临床疗效及安全性。方法回顾性选取2020年1月至2022年1月海南医学院第一附属医院TEN患者20例,均给予rhTNFR:Fc治疗。治疗21 d后,记录TE... 目的探讨重组人Ⅱ型肿瘤坏死因子受体-抗体融合蛋白(rhTNFR:Fc)治疗中毒性表皮坏死松解症(TEN)患者的临床疗效及安全性。方法回顾性选取2020年1月至2022年1月海南医学院第一附属医院TEN患者20例,均给予rhTNFR:Fc治疗。治疗21 d后,记录TEN患者临床疗效,比较治疗前与治疗后不同时段(治疗后7、14、21 d)的药疹面积和严重程度指数(DASI)评分[DASI评分平均值,50%DASI(DASI50)、75%DASI(DASI75)、90%DASI(DASI90)所占比例]、血清肿瘤坏死因子α(TNF-α)水平、体温下降时间、皮疹控制时间、住院时间及药物治疗的安全性。结果治疗21 d后,TEN患者中,显效18例(90.00%),有效2例(10.00%)。TEN患者治疗后7、14、21 d的DASI评分分别为(30.44±5.68)、(5.28±2.31)、(2.04±1.12)分,均明显低于治疗前[(52.34±7.45)分],差异均有统计学意义(P<0.05)。相较治疗前、治疗7 d、治疗14 d,治疗21 d后的DASI50(100.00%)、DASI75(100.00%)、DASI90(90.00%)的改善比率最高,差异均有统计学意义(P<0.05)。TEN患者治疗后7、14、21 d的血清TNF-α水平分别为(22.73±5.58)、(15.99±4.60)、(4.44±1.10)pg/mL,均低于治疗前[(33.63±17.36)pg/mL],差异均有统计学意义(P<0.05)。TEN患者体温下降时间为(2.49±0.81)d,皮疹控制时间为(5.19±1.90)d,住院时间为(11.92±4.20)d。治疗期间患者未出现终止治疗或失访,均未出现急性不良反应,随访期间病情未见复发,定期复查结果显示并无合并症、活动性肝炎与结核疾病等。结论rhTNFR:Fc作为治疗TEN疾病的药物其疗效随治疗时间延长而提高,可降低血清TNF-α水平与DASI评分,且安全性较高。 展开更多
关键词 重组人Ⅱ型肿瘤坏死因子受体-抗体融合蛋白 中毒性表皮坏死松解症 生物制剂 临床疗效 安全性
下载PDF
猪圆环病毒3型Cap蛋白单克隆抗体的制备及阻断ELISA检测方法的建立 被引量:1
17
作者 张宝戈 黄雅琴 +2 位作者 蔡金双 朱晨光 李玉峰 《畜牧兽医学报》 CAS CSCD 北大核心 2024年第3期1170-1178,共9页
旨在建立检测猪圆环病毒3型(PCV3)抗体的阻断ELISA方法,本研究利用原核表达的PCV3Cap重组蛋白免疫BALB/c小鼠制备获得了一株分泌阻断效果良好抗体的杂交瘤细胞株2E6。以重组Cap蛋白作为包被抗原,以辣根过氧化物酶(HRP)标记的2E6单克隆... 旨在建立检测猪圆环病毒3型(PCV3)抗体的阻断ELISA方法,本研究利用原核表达的PCV3Cap重组蛋白免疫BALB/c小鼠制备获得了一株分泌阻断效果良好抗体的杂交瘤细胞株2E6。以重组Cap蛋白作为包被抗原,以辣根过氧化物酶(HRP)标记的2E6单克隆抗体作为检测抗体,经条件优化后建立了一种检测PCV3抗体的阻断ELISA方法。用建立的阻断ELISA方法检测50份临床阴性血清,计算阻断率(PI)的临界值,以此来确定该方法的判定标准:当PI≤28.30%时,判定结果为阴性;当PI≥35.05%时,判定结果为阳性;当28.30%<PI<35.05%时,判定为可疑,重复一次试验后如果结果仍为可疑,则判定为阳性。特异性试验表明该方法与猪圆环病毒2型(PCV2)、猪伪狂犬病病毒(PRV)、猪繁殖与呼吸综合征病毒(PRRSV)以及猪瘟病毒(CSFV)的阳性血清均无交叉反应;敏感性试验表明其检测效价可达到1:128;重复性试验表明批内与批间的变异系数均小于10%;符合性检验表明该方法与PCV检测金标准免疫过氧化物酶单层试验(IPMA)比对的Kappa值达0.9,具有高度的一致性。综上所述,本研究建立的阻断ELISA方法具有良好的特异性与较高的符合率,可用于后期进行PCV3抗体的检测,为PCV3的流行病学调查与临床诊断提供技术支持。 展开更多
关键词 猪圆环病毒3型 Cap重组蛋白 单克隆抗体 阻断ELISA
下载PDF
类鼻疽菌Ⅲ型分泌系统BipD蛋白的重组表达及其免疫学特性研究
18
作者 南栋琪 文远 +7 位作者 陈建高 饶承龙 吴潘 张子元 王施韦 闫晶敏 李倩 毛旭虎 《陆军军医大学学报》 CAS CSCD 北大核心 2024年第15期1713-1720,共8页
目的重组表达类鼻疽菌Ⅲ型分泌系统BipD蛋白,制备多克隆抗体并鉴定其免疫学性质。方法利用pET-28a表达系统在Eschericahia coli(E.coli)BL21(DE3)中重组表达BipD蛋白,通过His Trap亲和层析纯化获得rBipD蛋白,免疫BALB/c小鼠获得rBipD多... 目的重组表达类鼻疽菌Ⅲ型分泌系统BipD蛋白,制备多克隆抗体并鉴定其免疫学性质。方法利用pET-28a表达系统在Eschericahia coli(E.coli)BL21(DE3)中重组表达BipD蛋白,通过His Trap亲和层析纯化获得rBipD蛋白,免疫BALB/c小鼠获得rBipD多克隆抗体;分别用兔抗类鼻疽菌血清和类鼻疽菌感染阳性患者血清进行Western blot实验检测rBipD的免疫反应性;通过Western blot及免疫荧光染色检测鉴定rBipD的免疫原性;以rBipD建立间接ELISA检测临床类鼻疽患者血清抗体。结果成功构建pET-28a-BipD重组质粒并转化至E.coli BL21(DE3)中诱导表达、纯化获得了相对分子质量约为36×10^(3)的rBipD蛋白,纯度约为95.4%。rBipD蛋白具有良好的免疫原性和免疫反应性。免疫小鼠可产生特异性抗体,制备鼠抗rBipD多克隆抗体,效价达1∶512000。5.0μg/mL rBipD蛋白可与类鼻疽患者血清发生免疫反应,而不与结核患者血清发生免疫反应,差异有统计学意义(P<0.01)。结论成功制备了具有免疫学活性的rBipD蛋白及其多克隆抗体,为其用于临床免疫学诊断和类鼻疽菌感染免疫机制研究提供了良好的工具。 展开更多
关键词 类鼻疽伯克霍尔德菌 BipD蛋白 重组表达 抗体制备
下载PDF
重组SARS-CoV-2 Omicron变异株S1蛋白的表达及免疫原性评价
19
作者 卢慧敏 王梓豪 +5 位作者 边成 王晓辉 李吉翠 马绍辉 褚嘉祐 杨昭庆 《医学研究杂志》 2024年第8期126-131,共6页
目的构建表达新型冠状病毒(SARS-CoV-2)Omicron BA.1变异株S1蛋白的真核表达载体,在CHO-K1细胞中进行表达,评价其免疫原性,并对佐剂和抗原剂量进行研究。方法构建重组质粒UCOE-Omi-S1,转染至CHO-K1工程细胞中进行表达和纯化,通过SDS-PAG... 目的构建表达新型冠状病毒(SARS-CoV-2)Omicron BA.1变异株S1蛋白的真核表达载体,在CHO-K1细胞中进行表达,评价其免疫原性,并对佐剂和抗原剂量进行研究。方法构建重组质粒UCOE-Omi-S1,转染至CHO-K1工程细胞中进行表达和纯化,通过SDS-PAGE和Western blot法实验鉴定重组S1蛋白。将BALB/c小鼠随机分为12组,即PBS组,无佐剂组(高、中、低剂量组),铝盐佐剂对照组,MF59佐剂对照组,铝盐佐剂实验组(高、中、低剂量组),MF59佐剂实验组(高、中、低剂量组),将按照分组要求配比的溶液经小鼠肌内注射3次,间隔14天,每2周尾静脉采血,末次免疫30天后取血分离血清,酶联免疫吸附试验(enzyme-linked immunosorbent assay,ELISA)检测血清抗体水平,并和SARS-CoV-2原型株的假病毒和SARS-CoV-2 Omicron BA.1变异株的假病毒进行假病毒中和实验。结果目的蛋白在CHO-K1工程细胞表达后可分泌到培养上清中,在相对分子质量约为70kDa处可见1条特异性蛋白表达条带,经Western blot法鉴定为Omi-S1蛋白。铝盐佐剂组和MF59佐剂组免疫诱导效果优于无佐剂组,铝盐佐剂组和MF59佐剂组免疫诱导效果相差不大,但MF59佐剂起效快;高、中、低剂量组的免疫诱导效果在第8周相差不大,但高剂量组起效快。假病毒中和实验表明,MF59佐剂实验组针对Omicron变异株的中和抗体水平高于铝盐佐剂组。结论本研究所构建的Omicron S1重组蛋白免疫原性良好,在小鼠体内产生了良好的体液免疫应答,并诱导高水平的对抗SARS-CoV-2假病毒的中和抗体,对佐剂和抗原剂量初步研究,结果显示,10μg以下抗原剂量搭配MF59佐剂可获得较好的免疫效果。本研究为SARS-CoV-2变异株的重组蛋白疫苗的研制提供了实验基础。 展开更多
关键词 SARS-CoV-2 重组蛋白疫苗 佐剂 假病毒 中和抗体
下载PDF
免疫鸡血清新城疫病毒抗体水平与攻毒保护效果的相关性研究
20
作者 沈欣悦 刘梅 +3 位作者 李建梅 俞燕 范建华 戴亚斌 《中国动物传染病学报》 CAS 北大核心 2024年第1期96-103,共8页
为了明确不同血清新城疫病毒(NDV)抗体水平对鸡的保护效果,本研究采用重组NDV灭活疫苗(A-Ⅶ株)对鸡进行免疫,通过交叉血凝抑制(HI)试验比较A-Ⅶ株与La Sota株间的血清学差异,并采用NDV标准强毒株F_(48)E_(8)和基因Ⅶ型强毒株JSC0804对... 为了明确不同血清新城疫病毒(NDV)抗体水平对鸡的保护效果,本研究采用重组NDV灭活疫苗(A-Ⅶ株)对鸡进行免疫,通过交叉血凝抑制(HI)试验比较A-Ⅶ株与La Sota株间的血清学差异,并采用NDV标准强毒株F_(48)E_(8)和基因Ⅶ型强毒株JSC0804对不同抗体水平免疫鸡进行了攻毒保护试验。结果显示:抗A-Ⅶ株血清用A-Ⅶ株抗原测得的平均HI抗体效价显著高于La Sota株抗原(P<0.01),约高1.5log2,而抗La Sota株血清用La Sota株抗原测得的平均HI抗体效价稍高于A-Ⅶ株抗原,但无显著差异(P>0.05),表明2种疫苗株存在一定的血清学差异。在试验条件下,所有免疫鸡经点眼滴鼻攻毒后均未表现明显临床症状,但存在不同程度的排毒现象,排毒率总体上随抗体效价升高呈逐渐下降趋势。A-Ⅶ株疫苗免疫鸡血清HI抗体效价分别达≥13log_(2)和≥14log_(2)时可完全阻止F_(48)E_8株和JSC0804株排毒。免疫鸡排毒一般仅限于攻毒后5 d内。本研究阐明了免疫鸡的抗体水平和免疫保护效果的相关性,为新城疫免疫控制提供了依据。 展开更多
关键词 新城疫 重组新城疫病毒灭活疫苗(A-Ⅶ株) 抗体效价 保护效果 排毒 相关性
下载PDF
上一页 1 2 39 下一页 到第
使用帮助 返回顶部