王一飞
个性化签名
- 姓名:王一飞
- 目前身份:
- 担任导师情况:
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学术头衔:
博士生导师
- 职称:-
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学科领域:
岩土力学
- 研究兴趣:
王一飞,1989年中科院昆明植物研究所硕士毕业,1995年7月华中理工科技大学毕业,获医学博士学位,1997年博士后在河南医科大学博士后流动站出站,1997年至1999年在暨南大学生物工程研究所工作,1999年赴日本国家融合技术研究所进修,2001年至2005年任暨南大学生物医药研究开发基地技术总监,2006年至今任暨南大学生物医药研究开发基地主任。
担任基因工程药物国家工程中心学术带头人、教育部基因组药物工程研究中心副主任、广东省生物工程药物重点实验室副主任,兼任广东省生物医学工程学会生物工程及生物传感器专业委员会主任、广东生物医学工程学会常务理事、广州市生物工程学会常务理事等职。长期担任广州市科技局、广东省科技厅科技发展规划、重大科技计划的咨询专家和项目审评专家。
先后参与和主持了国家十五"863"项目,广东省、广州市科技项目共二十余项,学校"211工程"二级项目负责人。在国内、外发表论文70余篇,主编《中药与肿瘤免疫》和《肿瘤免疫研究进展》专著2部,参编专著1部。已申请国家发明专利5项。获河南省教委科技进步奖二等1项,河南省科委科技进步三等奖1项。
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主页访问
1976
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关注数
0
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成果阅读
864
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成果数
13
王一飞, Chuan-hai Zhang a, b, f, Jia-hai Lu c, **, Yi-fei Wang a, Huan-ying Zheng e, Sheng Xiong a, Mei-ying Zhang a, Xin-jian Liu a, Jiu-xiang Li a, Zhuo-yue Wan e, Xin-ge Yan e, Shu-Yuan Qi f, Zhiyong Cuig, Biliang Zhang d, g, *
C.-h. Zhang et al./Vaccine23(2005)3196-3201,-0001,():
-1年11月30日
The immunogenicity of a candidate-inactivated vaccine prepared from SARS-CoV F69 strain was evaluated in Balb/c mice. Potent humoral immune responses were induced under the elicitation of three times of immunizations at 2-week intervals with this vaccine, combined with three types of adjuvants (Freunds adjuvant, Al(OH)3 adjuvant and CpG adjuvant). Titers of specific IgG antibodies in three test groups all peaked in the sixth week after first vaccination, but significant differences existed in the kinetics of specific IgG antibody levels. The strong neutralizing capacity exhibited in micro-cytopathic effect neutralization tests indicated the specific antibodies are protective. Western blot assay further demonstrated the specificity of the induced serum antibodies.
SARS coronavirus (, SARS-CoV), , Inactivated vaccine, Immunogenicity, Balb/, c m]ouse
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王一飞, ZHANG Chuan-hai, WANG Yi-fei, LIU Xin-jian, LU Jia-hai, QIAN Chui-wen, WAN Zhuo-yue, YAN Xin-ge, ZHENG Huan-ying, ZHANG Mei-ying, XIONG Sheng, LI Jiu-xiang and QI Shu-yuan
Chin Med J 2005;18(6):493-496,-0001,():
-1年11月30日
cepharanthine
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【期刊论文】Immunogenicity of SARS inactivated vaccine in BALB/c mice
王一飞, Sheng Xiong a, Yi-Fei Wang a, *, Mei-Ying Zhang a, Xin-Jian Liu a, Chuan-Hai Zhang a, Shi-Sheng Liu a, Chui-Wen Qian a, Jiu-Xiang Li a, Jia-Hai Lu b, Zhuo-Yue Wan c, Huan-Yin Zheng c, Xin-Ge Yan c, Min-Jie Meng d, Jiang-lin Fan e
S. Xiong et al./Immunology Letters 95(2004)139-143,-0001,():
-1年11月30日
Severe acute respiratory syndrome (SARS) is a serious infectious threat to public health. To create a novel trial vaccine and evaluate its potency, we attempted to generate a SARS inactivated vaccine using SARS coronavirus (SARS-CoV) strain F69 treated with formaldehyde and mixed with Al(OH)3. Three doses of the vaccine were used to challenge three groups ofBALB/c mice.We found that the mice exhibited specific IgM on day 4 and IgG on day 8. The peak titers of IgG were at day 47 in low-dose group 1∶19,200) and high-dose group (1∶38,400) whereas in middle-dose group (1∶19,200), the peak was at day 40. On day 63, the IgG levels reached a plateau. Neutralization assay demonstrated that the antisera could protect Vero-E6 cells from SARS-CoVs infection. Analysis of the antibody specificity revealed that the mouse antisera contained a mixture of antibodies specifically against the structure proteins of SARS-CoV. Furthermore, the mouse antisera conferred higher amount of antibodies against protein N, polypeptide S4 and S2 than those of proteins M and 3CL. These findings suggest that the inactivated SARS-CoV could preserve its antigenicity and the inactivated vaccine can stimulate mice to produce high levels of ntibodies with neutralization activity. Results also suggest that polypeptides originating from protein N or S might be a potential target for the generation of a recombinant SARS vaccine. © 2004 Elsevier B.V. All rights reserved.
SARS, Inactivated vaccine, IgG antibody, Neutralization
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王一飞, ZHANG Chuan-hai*, GUO Zhong-min*, ZHENG Huan-ying*, LU Jia-hai*, WANG Yi-fei*, YAN Xin-ge*, ZHAO Yong, DU Xiong-wei, ZHANG Xin, FANG Ling, LING Wen-hua, QI Shu-yuan, YU Xin-bing, and ZHONG Nan-shan
Chinese Medical Journal 2004; 117(11): 1625-1629,-0001,():
-1年11月30日
Background The etiologic agent of severe acute re spiratory syndrome (SARS) ha s been confirmed to be a novel coronavirus (CoV), namely SARS2CoV1 Developing safe and effective SARS2CoV vaccine s is e ssential for us to prevent the po ssible reemergence of it s epidemic1 Previous experience s indicate that inactivated vaccine is conventional and more hopeful to be succe ssfully developed1 Immunogenicity evaluation of an experimental inactivated SARS2CoV vaccine in rabbit s wa s conducted and reported in this paper1 Methods The large2scale cultured SARS2CoV F69 strain wa s inactivated with 014 % formaldehyde and purified, then used a s the immunogen combined with Freund’s adjuvant1 Eight adult New Zealand rabbit s were immunized four time s with this experimental inactivated vaccine1 Twelve set s of rabbit serum were sampled from the third day to the seventy2fourth day after the first vaccination1 The titers of specific anti-SARS2CoV IgG antibody were determined by indirect enzyme2linked immuno sorbent a ssay, and the neutralizing antibody titers were detected with micro2cytopathic effect neutralization te st1 Results Rapid and potent humoral immune re sponse s were induced by the inactivated SARS2CoV vaccine in all the eight te st rabbit s1 Titers of both specific IgG antibody and neutralizing antibody peaked at about six weeks after first vaccination, with the maximum value of 1∶81 920 and 1∶20 480,re spectively1 After that, serum antibody levels remained at a plateau or had a slight decrea se, though two boo sters were given in the succedent 4 to 5 weeks1 Cro ss neutralization re sponse existed between SARS2CoV F69 strain and Z22Y3 strain1 Co ncl us i o ns The inactivated SARS2CoV vaccine made from F69 strain owns strong immunogenicity, and the cro ss neutralization re sponse between the two different SARS2CoV strains give s a hint of the similar neutralizing epitope s, which provide stable ba se s for the development of inactivated SARS2 CoV vaccine s1
severe acute respiratory syndrome
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【期刊论文】pET15b-EGFP原核表达载体的构建及其表达和纯化
王一飞, 董晓, 王家宁黄永章郭凌郧
郧阳医学院学报,2005,24(6):321~325,-0001,():
-1年11月30日
目的:利用中间载体pGEM-T easy vector构建表达型载体pET15b-EGFP,在大肠杆菌中高效表达可溶性蛋白EGFP并将其纯化。方法:以质粒pLEGFP-C1为模板采用聚合酶链反应(PCR)的方法特异性扩增EGFP cD-NA序列,利用Taq DNA聚合酶的非模板依赖性末端转移酶活性催化PCR扩增的EGFP序列和三磷酸脱氧腺苷(dATP)反应,在EGFP序列两3’端加“ ,将纯化后的EGFP序列与中间载体pGEM-T easy vector连接后转化感受态大肠杆菌DH5ct,经蓝白筛选,挑取白色单菌落进行扩增、酶切鉴定,正确重组的质粒命名为pGEM-T-EGFP用XhoI和BamH1分别双酶切pGEM-T-EGFP和pET15b,低熔点琼脂糖回收EGFP eDNA和线性化的pET15b片段后将两片段相连,转化感受态大肠杆菌DH5ct并对重组质粒进行酶切鉴定和DNA测序,获取正确重组的表达型质粒并命名为pET15b-EGFP。将正确重组的质粒pET15b-EGFP转化感受态大肠杆菌BL21(DE3),用异丙基p-D-半乳糖苷(IPTG)诱导表达。利用表达蛋白N端的组氨酸“标签”(His-tag)进行N一树脂柱亲和层析纯化,SDS-PAGE和Western blotting鉴定纯化蛋白质。结杲:经测序证实重组质粒pET15b-EGFP中的EGFP eDNA序列与从Clontech公司购买的质粒pLEGFP-C1中的EGFP eDNA序列完全一致,从而成功构建了表达型重组质粒pET15b-EGFP。pET15b-EGFP转化感受态大肠杆菌BI21(DE )并经诱导后得到高效表达,表达量可达66me,/100ml菌液,SDS-PAGE和Western blotting显示纯化蛋白为目的蛋白EGFP。结论:表达型重组质粒pET15b-EGFP的成功构建和表达、纯化为细胞穿透肽-EGFP融合蛋白穿透细胞能力的研究奠定了基础。
TA克隆载体, 增强型绿色荧光蛋白(, EGFP), , 重组质粒, 蛋白表达, 蛋白纯化
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王一飞, Sheng Xiong, Yi-Fei Wang, Xiang-Rong Ren, Bing Li, Mei-Ying Zhang, Yong Luo, Ling Zhang, Qiu-Ling Xie, Kuan-Yuan Su
J Gastroenterol 2005; 11(7): 1077-1082,-0001,():
-1年11月30日
ficial multidomain protein, was selected as the model molecule of complex protein with 2 disulfide bonds. A BbFGF-producing plasmid, pJN-BbFGF, and a HBscFv producing-plasmid, pQE-HBscFv, were constructed and transformed into E. coli strains BL21(DE3) and M15[pREP4] respectively. At the same time, both plasmids were transformed into a reductase-deficient host strain, E. coli Origami(DE3). The 4 recombinant E. coli strains were cultured and the target proteins were purified. Solubility and bioactivity of recombinant BbFGF and HBscFv produced in different host strains were analyzed and compared respectively. RESULTS: All recombinant E. coli strains could efficiently produce target proteins. The level of BbFGF in BL21(DE3) was 15-23% of the total protein, and was 5-10% in Origami (DE3). In addition, 65% of the BbFGF produced in BL21(DE3) formed into inclusion body in the cytoplasm, and all the target proteins became soluble in Origami (DE3). The bioactivity of BbFGF purified from Origami(DE3) was higher than its counterpart from BL21(DE3). The ED50 of BbFGF from Origami(DE3) and BL21(DE3) was 1.6μg/L and 2.2μg/L, respectively. Both HBscFv formed into inclusion body in the cytoplasm of M15[pQE-HBscFv] or Origami[pQE-HBscFv]. But the supernatant of Origami [pQE-HBscFv] lysate displayed weak bioactivity and its counterpart from M15[pQE-HBscFv] did not display any bioactivity. The soluble HBscFv in Origami[pQE-HBscFv] was purified to be 1-2 mg/L and its affinity constant was determined to be 2.62×107 mol/L. The yield of native HBscFv refolded from inclusion body in M15[pQE-HBscFv] was 30-35 mg/L and the affinity constant was 1.98×107 mol/L. There was no significant difference between the bioactivity of HBscFvs refolded from the inclusion bodies produced in different host strains. CONCLUSION: Modification of the redox environment of E. coli cytoplasm can significantly improve the folding of recombinant disulfide-bonded proteins produced in it.
Escherichia coli, Recombinant protein expression, Disulfide bond, Solubility, bFGF, HBsAg, Single-chain Fv, Reductase deficient, Protein folding, Affinity constant
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【期刊论文】蓝谷牌鲨鱼口服液对2型糖尿病病人短期病情控制、症状影响研究
王一飞, 陈伟
中国卫生检验杂志,2005,15(10):1243~1245,-0001,():
-1年11月30日
目的:研究鲨鱼口服液对15名2型糖尿病病人短期病情控制、症状影响。方法:采用单盲、交叉对照等方法,对鲨鱼口服液受试者的血、尿常规,生化指标、血糖及糖化血清蛋白,以及临床症状等指标进行考察,观察时间为20 d。结果:患者在服用鲨鱼口服液可能改善糖尿病的症状,试验前10.7±6.4分,服用口服液后降为5.8±5.3,与安慰剂对比(P<0.05)。同时该试验品对糖尿病患者的肝、肾、心脏功能及血液学功能等无明显不良影响。结论:鲨鱼口服液在2型糖.尿病患者短期应用安全、可能改善症状、应用简便,但对血糖的影响仍需要长期观察。
2型糖尿病, 口服液, 研究
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王一飞, 钱垂文, 熊盛, 张关英, 陆家海, 张传海, 李久香, 万卓越, 郑焕英, 张庶民
现代免疫学,2005,25(2):152~154,-0001,():
-1年11月30日
为制备纯化的灭活SARS病毒,取广东省CDC提供的灭活SARS-CoV病谳培养液进行超滤,收集相对分子质量介于100000至500000之间的组分。并浓缩。对纯化过程中各组分进行抗原活性签定和蛋白质含量测定。并加佐剂后免疫动物,测定抗体免疫应答。结果表明:超滤后活性抗原蛋白质回收率7.6%,纯化倍数为9,抗原活性达到1∶480 免疫家免获得的血清ELISA效价达到1∶10000以上,中和抗体效价达到1∶2000以上。表明该纯化工艺能获得较高纯度的灭活SARS-CoV病毒颗粒。
SARS-CoV, 纯化, ELISA, 免疫应答
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【期刊论文】达NM23-HI/NDPK-A工程菌的遗传稳定性研究*
王一飞, 钱垂文, 黄立, 王一飞**, 熊盛, 张美英, 李雪玲
中国生物工程杂志,2005,25(2):35~38,-0001,():
-1年11月30日
目的:研究重组工程菌的遗传稳定性。方法:利用重组表达质粒pBVNM-HI转化宿主菌E.coli DI-LSa,筛选重组工程菌DH5a-pBVNM-HI。将新构建好的重组工程菌在无选择压力的条件下进行连续传代培养,比较菌落在LB(-)和LB(+)培养基上的生长状况,并对传代菌株目标蛋白的表达情况以及质粒数量和目的基因DNA进行电泳鉴定。结果:重组工程菌连续传代5O次中,在LB(-)和LB(+)培养基上的生长状况相同,目标蛋白表达量无显著差异,质粒数量及目的基因DNA结构稳定。结论:重组工程菌DH5a-pBVNM.HI具有良好的遗传稳定性。
pBVNM-HI DH5a-pBVNM-HI 遗传稳定性
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【期刊论文】灭活SARS冠状病毒疫苗对小鼠脏器影响的实验研究*
王一飞, 杜彬, 钟雪云, 熊盛, 张传海, 刘新建, 刘石生, 张美英, 李久香, 陆家海, 万卓越, 鄢心革, 郑焕英, 范江林
中国病理生理杂志,2004,20(7):1187~1189,-0001,():
-1年11月30日
目的:探讨灭活SAILS冠状病毒疫苗对小鼠是否有损伤作用。方法:用灭活SARS冠状病毒分别免疫BALB/c和C57BL/6小鼠;ELISA法检测小鼠血清中抗SARS冠状病毒抗体水平;免疫8周后取小鼠各脏器,观察其病理形态学改变。结果:免疫8d后,小鼠血清中就可检测到特异的Ig,M、IgG抗体;组织切片染色观察,可见少数小鼠肺及肝组织出现轻度损伤,其他脏器未见病变。结论:SAILS冠状病毒灭活疫苗可诱导小鼠产生特异性抗体,同时没有造成严重的器官损伤。这将为SARS疫苗进入临床研究打下基础。
严重急性呼吸综合征; 冠状病毒; 疫苗, 灭活; 小鼠
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