洪岸
细胞生长因子研究、干细胞与组织工程研究
个性化签名
- 姓名:洪岸
- 目前身份:
- 担任导师情况:
- 学位:
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学术头衔:
博士生导师
- 职称:-
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学科领域:
生物化学
- 研究兴趣:细胞生长因子研究、干细胞与组织工程研究
洪岸,女,医学博士,研究员,博士生导师,暨南大学生物工程研究所所长,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,广东省人体生物组织工程学会常务理事,广东省免疫学会理事,广州生物工程学会理事,中国生物工程学会会员,中国药学会会员。
近十年来,主要从事基因工程技术及其新药的研究和开发,围绕细胞生长因子及其新药研究开展了重组细胞生长因子及其受体的基因表达、调控研究,重组细胞生长因子及其受体基因突变、蛋白修饰研究,生长因子受体及其突变体的研究,生长因子与其受体相互作用研究以及生长因子与化学合成小分子相互作用研究等系统性的研究,建立了细胞生长因子/受体研究体系和重组蛋白质新药研究开发体系。发现了多个新药物,已成功开发了两个基因工程新药(原一类),在国内该领域走在前列。该研究依托的生物工程研究所成功申报成为广东省生物工程药物重点实验室、教育部基因组药物工程研究中心和国家基因工程药物工程研究中心。多年作为广州市科技局,广东省科技厅科技发展规划、重大科技计划,国家自然科学基金的项目审评专家。在近两年由细胞生长因子研究延伸到干细胞与组织工程研究领域,开展了转生长因子基因干细胞研究、生长因子与材料的复合技术及多元生长因子的协同作用研究等。
上述研究工作得到国家"863"项目、国家"十五"重大专项、国家"973"项目子项目、国家自然科学基金项目、广东省"十五"重大专项、广东省基金重点项目、广东省团队项目、广州市科技攻关重大项目、"211工程"项目等资助,近五年累计纵向科研资助1110万元。横向科研经费1500万。申请发明专利10余项,已授权4项。获奖5项,其中包括教育部提名国家科技进步奖、广东省自然科学奖等奖项。发表论文80余篇。
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成果阅读
663
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成果数
9
洪岸
生理学进展,1995,26(3):240~242,-0001,():
-1年11月30日
时由Morris水迷宫分得的青年、老年记忆正常和记忆减退鼠的脑组织分别进行突触,AChE纤维、突触素、小白蛋白神经元以及突触体钙离子浓度([Ca2+])、膜流动性的定量分析。结果表明老年记忆减退鼠新皮质、海马结构突触素含量、突触、胆碱能纤维、小白蛋白阳性神经元密度及突触体膜流动性较老年记忆正常和青年鼠明显降低。[Ca2+]明显增加。老年记忆正常鼠与青年鼠各项均乇显著差异。本研究提示各研究指标的异常与老年学习记忆减退密切相关。
老年学习记忆减退, 突触素, 小白蛋白, 突触体膜流动性, 钙离子
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【期刊论文】碱性成纤维细胞生长因子对小鼠脾脏细胞凋亡的抑制作用*
洪岸, 许艳丽, 李校坤, 林剑, 刘树铮
中国病理生理杂志,2000,16(2):131~134,-0001,():
-1年11月30日
目的和方法:X射线照射结合无血清培养诱导小鼠脾细胞凋亡,采用流式细胞术定量分析碱性成纤维细胞生长因子(bFGF)对睥细胞凋亡的抑制作用,井采用淋巴细胞转化试验[3H]-TdR掺人法检测bFGF对脾细胞的增殖效应。结果:小鼠经过不同剂量X射线照射、脾细胞再经过不同时程无血清培养,都出现了典型的亚二倍体Ap峰,1μg/mL和2μg/mL的bFCF均可降低Ap峰中的细胞数(细胞凋亡率)。并能显著增强睥细胞的增殖反应能力。结论:bFCF可抑制外周免疫器官脾细胞的凋亡、井促进其增殖。增强机体免疫力。
成纤维细胞生长因子、碱性, 脾, 细胞, 小鼠
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洪岸, CHEN Xiao-Jia, SUN Fen-Yong, XIE Qiu-Ling, LIAO Mei-De , ZHANG Ling, LI Zhi-Ying, HONG An, LIN Jian
Chen XJ et al/Acta Pharmacol Sin, 2002, 9, 23(9): 782-786,-0001,():
-1年11月30日
AIM: To obtain high-level expression of nonfusion recombinant human basic fibroblast growth factor (rhbFGF). METHODS: hbFGF cDNA was prepared from the total RNA of embryonic brain tissue. As a template, the obtained gene was used to clone nonfusion rhbFGF. New primers were employed to alter the translation initiation region (TIR) and reduce the G+C content through nucleotide change. Using pET-3C as vector, the cloned rhbFGF was expressed in BL21 (DE3). RESULTS: rhbFGF was expressed in E coli up to 30% of the total cellular protein. Cation exchange and heparin affinity chromatography were employed to purify the target protein from the supernatant of bacteria lysate. The bioactivity of the purified rhbFGF was identical with the standard bFGF. CONCLUSION: Modification of TIR is an effective means to increase nonfusion expression rate of recombinant proteins, such as rhbFGF, in E coli.
basic fibroblast growth factor, molecular cloning, gene expression
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【期刊论文】Update on therapeutic neovascularization
洪岸, Yihai Cao a, *, An Hong b, Henny Schulten c, Mark J. Post c
y. Cao et al./Cardiovascular Research 65(2005)639-648,-0001,():
-1年11月30日
Therapeutic neovascularization for cardiovascular ischemia is a promising avenue in spite of disappointing early clinical trial results. The concept of three different mechanisms of neovascularization has served to define potential therapeutic targets such as vascular remodeling and stem cell recruitment, but it is anticipated that this will lose significance as the pleiotropic nature of angiogenic cytokines becomes fully understood. With the rapidly growing body of data on growth factors and pro-angiogenic strategies, approaches will emerge that are more effective than the ones that have been tested clinically thus far. Combinations of growth factors, for instance to stabilize vessels, or growth factors combined with cell transplants deserve more attention but will make the design of preclinical and clinical studies increasingly complex. Recent developments suggest that when using the appropriate dose and treatment regimens, even single growth factor therapy can result in stable and functional vessels. Whether gene therapy or protein therapy will be optimal for this purpose depends mainly on technical developments in vector design and production and on progress in the engineering of slow release matrix formulations for proteins. With the increasing complexity of therapeutic strategies, it remains imperative that these approaches are rationally based on fundamental and preclinical data.
Ischemia, Neovascularization, Angiogenesis, Growth factors
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洪岸, Rong-jie Yu, Qiu-ling Xie, Yun Dai, Yuan Gao, Tian-hong Zhou, An Hong *
PEPTIDES 27(2006)1359-1366,-0001,():
-1年11月30日
In order to obtain the recombinant VPAC2 agonist efficiently by intein-mediated single column purification, a gene encoding 32-amino acids peptide was designed, synthesized and cloned into Escherichia coli expression vector pKYB. The recombinant vector pKY-ROM was transferred into E. coli ER2566 cells and the target protein was over-expressed as a fusion to the N-terminus of a self-cleavable affinity tag. After the rMROM-intein-CBD fusion protein was purified by chitin-affinity chromatography, the self-cleavage activity of the intein was induced by β-mercaptoethanol and the rMROM with the homogeneity over 95% was released from the chitin-bound intein tag. The recombinant linear rMROM competitively displaced [125I] PACAP38 on VPAC2 with a half-maximal inhibitory concentration (IC50) of 60±5 nM, whereas the IC50 of rMROM at human VPAC1 was observed up to 10 mMand no binding was detected at PAC1. rMROM stimulated the cAMP accumulation in Chinese hamster ovary (CHO) cells expressing the human VPAC2 with a half-maximal stimulatory concentration (EC50) of 0.6 nM, which was 500-fold less potent at VPAC1and had no activity on PAC1. An efficient production procedure of a novel recombinant VPAC2-selective agonist was established.
Intein Recombinant VPAC2 agonist Purification
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洪岸, Ju Wang a, *, An Hong a, **, Jin-song Ren b, Fen-Yong Sun a, Ying-jiao Shi a, Kan Liu a, Qiu-Ling Xie a, Yun Dai a, Zhi-ying Li a, Yu Chen c
J. Wang et al./Journal of Biotechnology 121(2006)442-447,-0001,():
-1年11月30日
Fibroblast growth factor-2 (FGF-2) is a multifunctional polypeptide that affects many cellular functions and phenomena. The wild-type recombinant human fibroblast growth factor rhFGF-2W and the mutant C78SC96S rhFGF-2M were expressed in Escherichia coli and their products were purified. The results by the means of fluorescence spectroscopy and CD spectrums, suggested that due to its decreased hydrophobicity rhFGF-2 is not deposited as an inclusion body. The mitogenic activity of the expressed rhFGF-2M on 3T3 fibroblasts was shown to be 10-fold more than the expressed rhFGF-2W of which the biological activity was a little less than that of the standard rhbFGFW, indicating that the increased biological activity was due to the change of its secondary structure, dimerization and affinity binding to FGF receptor (FGFR).
rhFGF-2, Mutation, Fluorescence spectroscopy, Mitogenic activity
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洪岸, 孙晋华, 洪岸*, 张玲, 孙奋勇, 宋照熙
微生物学报,2003,43(4):448~452,-0001,():
-1年11月30日
包涵体的形成与外源基因在大肠杆菌中的表达量高度相关,在适当的范围内,降低hbFCF在表达宿主BL21(DE3)plysS中的表达,成为实现高可溶性表达的关键。在不改变氨基酸序列的条件下,对hbFCF高表达菌株突变重组子起始密码ATC下游前3个密码子的摇摆碱基进行随机回复突变,共有7种组合,合成引物PCR扩增后,克隆至表达载体pET-3e,将重组子转导BL21(DE3)plysS后,IPTC诱导表达,发现其中1株有较高可溶性和活性的菌株。可见部分降低外源蛋白的表达量可以避免与减少包涵体的形成。
可溶性表达,, 包涵体,, 人碱性成纤维细胞生长因子,, 大肠杆菌,, 回复突变
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洪岸, HUANG Ceng-sheng , WANG Ju , SUN Fen-yong , LI Zhi-ying , ZHANG Ling , HONG An #, LI Gui-sheng
中国病理生理杂志,2003,19(5):627~631,-0001,():
-1年11月30日
AIM: To investigate the condition necessay for high-level expression of basic fibrob-last growth factor (hbFGF) in Escherichia coli, adicistron system was constructed. METHODS:1he phage T7 gene φ10 N-tenninal sequence, present in the expression plasmid pET-3c, was incolporated into the first cistron, the coding region for bFCF was incorporated into the second, and boLh were under the control of a transcnption promotor recogruzed by the bacteriophage T7 RNA polymerase. RESULTS: Recambinants were transfonned into BI21(DE3) and induced by IPTG for expression, accumulation of up to 20% of the total ceU protein of bFGF were obtained, showing bioactivity indistingushable from standard protein. CONCLUSION: It suggested that such dicistron system was effective for enhancing hbFCF expression.
Dicistron, Fibroblast growth factor 2, Escherichia coli
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洪岸, 汪炬, 洪岸**, 孙奋勇, 谢秋玲
生物化学与生物物理进展,2003,30(5):772~777,-0001,():
-1年11月30日
采用两性离子胶体沉淀和乙醇沉淀相结合的粗提方法,经离子交换、疏水层析、亲和层析及凝胶过滤4个步骤有效地将人尿激肽原酶(hk-l)粗提物纯化,比活提高了1755倍,总得率为70%.用以慈菇蛋白酶抑制剂为配体的亲和层析纯化hk-l,效果理想,整个工艺路线适合产业化生产。纯化产物在SDS-聚丙烯酰胺凝胶电泳上为单带,高压液相色谱(HPLC)上为单峰,基质辅助激光解析电离飞行时间质谱测得分子质量为33 450u,等电聚焦测得p14.3附近,为含糖蛋白。同时测定了该酶的热稳定性和pH稳定性。纯化过程中同时分离得到另一种药用蛋白—人尿胰蛋白酶抑制剂(HUTI)。
人尿激肽原酶,, 人尿胰蛋白酶抑制剂,, 乙醇沉淀,, 胶体沉淀,, 离子交换,, 蔬水层析,, 亲和层析,, 慈菇蛋白酶抑制剂
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