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Preparation and application of immunoassay signal amplification antibody conjugates Scopus CSCD PKU
期刊论文 | 2023 , 43 (1) , 42-49 | China Biotechnology
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Abstract :

Objective: Chemical coupling agent and polylysine (PL) were used to conjugate goat anti-rabbit IgG and horseradish peroxidase (HRP) to increase the number of HRP connections on antibodies to amplify the sensitivity of immune detection, and the sensitivity was detected and compared through immunodetection applications, which is of great significance for subsequent immune diagnosis and biological research. Methods: Tylthioacetate (SATA) and Traut’s were optimized for the polymer HRP-PL, the feed ratio of Succinimidyl 4-[N-maleimidomethyl]cyclohexane-1-carboxylate (Sulfo-SMCC) was optimized with goat anti-rabbit IgG and HRP-PL, and the reaction ratio of the two was optimized. The conjugated IgG-PL-HRP and commercial secondary antibody were detected by immunoblot, ELISA and immunohistochemistry, respectively, and the amplification of conjugated IgG-PL-HRP was calculated. Results: When the molar ratio of PL to HRP was 1∶5, the molar ratio of HRP-PL to Traut’s was 1∶15, the molar ratio of goat anti-rabbit IgG to Sulfo-SMCC was 1∶30, the molar ratio of goat anti-rabbit IgG to HRP-PL was 1∶10, the reaction efficiency was high. In the immune spot test, the minimum detection limits of commercial secondary antibody and conjugate IgG-PL-HRP were 2.5 μg and 312.5 ng, respectively, and the maximum dilution times were 50 and 100 times, respectively. In ELISA experiment, the maximum dilution times were 5 000 and 20 000, times respectively. In the immunohistochemical experiment, the detection specificity and intensity of conjugated IgG-PL-HRP were higher than those of commercial secondary antibody. Conclusion: The primary amplification conjugate IgG-PL-HRP was successfully synthesized and its immune detection signal amplification was about 3 ~ 7 times that of the commercial secondary antibody. It is of great significance to the following immunodiagnosis and biological research. © 2023, China Biotechnology Press. All rights reserved.

Keyword :

Antibody conjugate Antibody conjugate Horseradish peroxidase Horseradish peroxidase Polylysine Polylysine Signal amplification Signal amplification

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GB/T 7714 Shen, Y. , Yao, B.-Y. , Yang, R.-N. et al. Preparation and application of immunoassay signal amplification antibody conjugates [J]. | China Biotechnology , 2023 , 43 (1) : 42-49 .
MLA Shen, Y. et al. "Preparation and application of immunoassay signal amplification antibody conjugates" . | China Biotechnology 43 . 1 (2023) : 42-49 .
APA Shen, Y. , Yao, B.-Y. , Yang, R.-N. , Kang, W.-B. , Lin, H.-Y. . Preparation and application of immunoassay signal amplification antibody conjugates . | China Biotechnology , 2023 , 43 (1) , 42-49 .
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免疫检测信号放大抗体偶联物的制备及应用 CSCD PKU
期刊论文 | 2023 , 43 (1) , 42-49 | 中国生物工程杂志
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目的:以聚赖氨酸(polylysine,PL)为骨架提高辣根过氧化物酶(horseradish peroxidase,HRP)与羊抗兔IgG的连接数量,比较几种化学偶联剂的偶联效果,通过免疫检测技术对其灵敏度进行检测和比较.方法:对HRP与PL、聚合物HRP-PL与N-琥珀酰-S-乙酰乙酸(N-succinimidyl-S-acetylthioacetate,SATA)和2-亚氨基硫烷(Traut's)两种试剂、羊抗兔IgG与琥珀酰亚胺基4-(N-马来酰亚胺甲基)环己烷-1-羧酸酯[succinimidyl 4-(N-maleimidomethyl)cyclohexane-1-carboxylate,Sulfo-SMCC]、活化后羊抗兔IgG及HRP-PL进行摩尔比的优化;对偶联物IgG-PL-HRP及商品化二抗分别进行斑点免疫印迹、ELISA和免疫组化,并计算偶联物IgG-PL-HRP的检测放大倍数.结果:当PL与HRP摩尔比为1∶5,HRP-PL与Traut's摩尔比为1∶15,羊抗兔IgG与Sulfo-SMCC摩尔比为1∶ 30,羊抗兔IgG与HRP-PL摩尔比为1:10时,反应效率较高;商品化二抗及偶联物IgG-PL-HRP在斑点免疫印迹实验中的最低检测限分别为2.5μg和312.5 ng,最大稀释倍数分别为50和100倍;在ELISA实验中的最大稀释倍数分别为5 000和20 000倍;在免疫组化实验中偶联物IgG-PL-HRP的检测特异性及强度均大于商品化二抗.结论:成功合成抗体偶联物IgG-PL-HRP,且免疫检测信号放大倍数为商品化二抗的3~7倍,这对后续免疫诊断及生物学的研究具有重要意义.

Keyword :

信号放大 信号放大 抗体偶联物 抗体偶联物 聚赖氨酸 聚赖氨酸 辣根过氧化物酶 辣根过氧化物酶

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GB/T 7714 沈妍 , 姚玢妍 , 杨若楠 et al. 免疫检测信号放大抗体偶联物的制备及应用 [J]. | 中国生物工程杂志 , 2023 , 43 (1) : 42-49 .
MLA 沈妍 et al. "免疫检测信号放大抗体偶联物的制备及应用" . | 中国生物工程杂志 43 . 1 (2023) : 42-49 .
APA 沈妍 , 姚玢妍 , 杨若楠 , 康文斌 , 林海英 . 免疫检测信号放大抗体偶联物的制备及应用 . | 中国生物工程杂志 , 2023 , 43 (1) , 42-49 .
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组合型嵌合抗原受体的构建及分析鉴定 CSCD PKU
期刊论文 | 2023 , 43 (06) , 12-19 | 中国生物工程杂志
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目的:构建靶向磷脂酰肌醇蛋白聚糖3(glypican-3,GPC3)的嵌合抗原受体(chimeric antigen receptor, CAR)及靶向上皮细胞黏附分子(epithelial cell adhesion molecule, EPCAM)嵌合抗原共刺激受体(chimeric antigen costimulatory receptor, CCR)共修饰的T细胞,并对其进行体外活性评估。方法:将EPCAM-CCR和GPC3-CAR基因片段克隆入慢病毒载体质粒,酶切、PCR和测序鉴定CCR+CAR pCDH重组载体。分离、激活和扩增人T细胞,利用慢病毒感染并筛选能够稳定表达该组合型嵌合抗原受体人T细胞。通过Western blot、流式细胞术(flow cytometry, FCM)验证CCR+CAR T细胞中CCR+CAR的表达,ELISA检测细胞因子IL-2、INF-γ、IL-4的分泌。结果:成功构建CCR+CAR pCDH慢病毒重组载体。成功分离、激活并扩增人T细胞。CCR+CAR pCDH慢病毒成功感染人T细胞,RT-PCR、Western blot检测也显示其成功表达目的蛋白,同时FCM分析显示CCR+CAR T细胞表面CCR+CAR的表达率为42%左右;ELISA检测CCR+CAR与表达GPC3和EPCAM的HepG2、Huh-7共培,其分泌更高水平的IL-2、INF-γ、IL-4,同时与人正常肝细胞L-02共培未发现CCR+CAR T细胞的有效激活。结论:成功获得组合型CCR+CAR T细胞,并初步探究其在肝癌细胞中的免疫效应,为后续动物体内免疫及实体瘤过继免疫治疗中防脱靶效应奠定基础。

Keyword :

上皮细胞黏附分子 上皮细胞黏附分子 嵌合抗原共刺激受体 嵌合抗原共刺激受体 嵌合抗原受体 嵌合抗原受体 磷脂酰肌醇蛋白聚糖3 磷脂酰肌醇蛋白聚糖3

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GB/T 7714 林海英 , 姚玢妍 , 于艺洁 et al. 组合型嵌合抗原受体的构建及分析鉴定 [J]. | 中国生物工程杂志 , 2023 , 43 (06) : 12-19 .
MLA 林海英 et al. "组合型嵌合抗原受体的构建及分析鉴定" . | 中国生物工程杂志 43 . 06 (2023) : 12-19 .
APA 林海英 , 姚玢妍 , 于艺洁 , 杨扬 . 组合型嵌合抗原受体的构建及分析鉴定 . | 中国生物工程杂志 , 2023 , 43 (06) , 12-19 .
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Construction, Analysis and Characterization of Combinvssatorial Chimeric Antigen Receptor Scopus CSCD PKU
期刊论文 | 2023 , 43 (6) , 12-19 | China Biotechnology
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To construct T cells encoding chimeric antigen receptor (CAR) targeting glypican-3 (GPC3) and chimeric antigen costimulatory receptor (CCR) targeting epithelial cell adhesion molecule (EPCAM), and analyze their activities in vitro. Methods: Gene fragments encoding EPCAM-CCR and GPC3- CAR was cloned into lentivirus vector plasmid. The recombinant CCR + CAR PCDH vector was identified by enzyme digestion, PCR and sequencing. T lymphocytes were isolated, activated and expanded. A stable combinatorial CAR modified cell line was generated using the lentivirus. Western blot, RT-PCR and flow cytometry (FCM) were used to verify the expression of CCR+ CAR in CCR+ CAR T cells. The secretion of cytokines IL-2, IL-4 and INF-y was detected by ELISA. Results: The CCR + CAR PCDH lentiviral recombinant plasmid was successfully constructed. Human T lymphocytes were successfully isolated, activated and expanded. Human T lymphocytes were successfully infected with CCR + CAR PCDH lentivirus. RT-PCR and Western blot showed that the target protein was successfully expressed, and FCM analysis showed that the expression rate of CCR + CAR on CCR + CAR T cells was about 42%. ELISA was used to detect CCR + CAR co-culture with HepG2 and Lu-7 expressing GPC3 and EPCAM, the secretion levels of IL-2, INF-y and IL-4 were higher, and the co-culture of human normal liver cells L-02 did not find effective activation of CCR+ CAR T cells. Conclusions: The combined CCR + CAR T cells were successfully obtained, and the immune effect of CAR T combined with the two antigen recognition signals was attempted in hepatocellular carcinoma cells. The result laid a foundation for the immune effects in the subsequent in vitro experiments, and provided a potentially novel approach to augment the off-target effect of CAR-T immunotherapy for solid tumors. © 2023, China Biotechnology Press. All rights reserved.

Keyword :

Chimeric antigen costimulatory receptor Chimeric antigen costimulatory receptor Chimeric antigen receptor (CAR) Chimeric antigen receptor (CAR) Epithelial cell adhesion molecule Epithelial cell adhesion molecule Glypican-3 Glypican-3

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GB/T 7714 Lin, H.-Y. , Yao, B.-Y. , Yu, Y.-J. et al. Construction, Analysis and Characterization of Combinvssatorial Chimeric Antigen Receptor [J]. | China Biotechnology , 2023 , 43 (6) : 12-19 .
MLA Lin, H.-Y. et al. "Construction, Analysis and Characterization of Combinvssatorial Chimeric Antigen Receptor" . | China Biotechnology 43 . 6 (2023) : 12-19 .
APA Lin, H.-Y. , Yao, B.-Y. , Yu, Y.-J. , Yang, Y. . Construction, Analysis and Characterization of Combinvssatorial Chimeric Antigen Receptor . | China Biotechnology , 2023 , 43 (6) , 12-19 .
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Expression, purification, and characterization of cell-permeable fusion antioxidant enzyme sensitive to matrix metalloproteinases-2/9 [基质金属蛋白酶-2/9 敏感型可跨膜融合抗氧化酶的表达、纯化和表征] Scopus CSCD PKU
期刊论文 | 2022 , 38 (9) , 3515-3527 | Chinese Journal of Biotechnology
SCOPUS Cited Count: 1
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Antioxidant enzymes fused with cell-penetrating peptides could enter cells and protect cells from irradiation damage. However, the unselective transmembrane ability of cell-penetrating peptide may also bring antioxidant enzymes into tumor cells, thus protecting tumor cells and consequently reducing the efficacy of radiotherapy. There are active matrix metalloproteinase (MMP)-2 or MMP-9 in most tumor cellular microenvironments. Therefore, a fusion protein containing an MMP-2/9 cleavable substrate peptide X, a cell-penetrating peptide R9, a glutathione S-transferase (GST), and a human Cu, Zn superoxide dismutase (SOD1), was designed and named GST-SOD1-X-R9. In the tumor microenvironment, GST-SOD1-X-R9 would lose its cell-penetrating peptide and could not enter tumor cells due to the cleavage of substrate X by active MMP-2/9, thereby achieving selected entering normal cells. The complete nucleotide sequence of SOD1-X-R9 was synthesized and inserted into the prokaryotic expression vector pGEX-4T-1. The pGEX4T-1-SOD1-X-R9 recombinant plasmid was obtained, and soluble expression of the fusion protein was achieved. GST-SOD1-X-R9 was purified by ammonium sulfate precipitation and GST affinity chromatography. The molecular weight of the fusion protein was approximately 47 kDa, consistent with the theoretical value. The SOD and GST activities were 2 954 U/mg and 328 U/mg, respectively. Stability test suggested that almost no change in either SOD activity or GST activity of GST-SOD1-X-R9 was observed under physiological conditions. The fusion protein could be partially digested by collagenase IV in solution. Subsequently, the effect of MMP-2/9 activity on transmembrane ability of the fusion protein was tested using 2D and 3D cultured HepG2 cells. Little extracellular MMP-2 activity of HepG2 cells was observed under 2D culture condition. While under the 3D culture model, the size and the MMP-2 activity of the HepG2 tumor spheroid increased daily. GST-SOD1-R9 proteins showed the same transmembrane efficiency in 2D cultured HepG2 cells, but the transmembrane efficiency of GST-SOD1-X-R9 in 3D cultured HepG2 spheres was reduced remarkably. This study provided a basis for further investigating the selectively protective effect of GST-SOD1-X-R9 against oxidative damage in normal cells. ©2022 Chin J Biotech, All rights reserved.

Keyword :

antioxidant enzyme antioxidant enzyme cell permeable cell permeable expression in Escherichia coli expression in Escherichia coli MMP-2/9 sensitive MMP-2/9 sensitive purification purification stability stability

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GB/T 7714 He, H. , Lin, L. , Li, L. et al. Expression, purification, and characterization of cell-permeable fusion antioxidant enzyme sensitive to matrix metalloproteinases-2/9 [基质金属蛋白酶-2/9 敏感型可跨膜融合抗氧化酶的表达、纯化和表征] [J]. | Chinese Journal of Biotechnology , 2022 , 38 (9) : 3515-3527 .
MLA He, H. et al. "Expression, purification, and characterization of cell-permeable fusion antioxidant enzyme sensitive to matrix metalloproteinases-2/9 [基质金属蛋白酶-2/9 敏感型可跨膜融合抗氧化酶的表达、纯化和表征]" . | Chinese Journal of Biotechnology 38 . 9 (2022) : 3515-3527 .
APA He, H. , Lin, L. , Li, L. , Wu, L. , Lin, H. , Pan, J. . Expression, purification, and characterization of cell-permeable fusion antioxidant enzyme sensitive to matrix metalloproteinases-2/9 [基质金属蛋白酶-2/9 敏感型可跨膜融合抗氧化酶的表达、纯化和表征] . | Chinese Journal of Biotechnology , 2022 , 38 (9) , 3515-3527 .
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基质金属蛋白酶-2/9敏感型可跨膜融合抗氧化酶的表达、纯化和表征 CSCD PKU
期刊论文 | 2022 , 38 (9) , 3515-3527 | 生物工程学报
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融合了跨膜肽的抗氧化酶可进入细胞,保护细胞免受放射损伤.然而跨膜肽的跨膜能力没有靶向性,其也可把抗氧化酶带入肿瘤细胞进而保护肿瘤细胞,降低放疗的效果.为此,根据多数肿瘤细胞微环境中存在活性基质金属蛋白酶(matrix metalloproteinase,MMP)-2或MMP-9的特点,在细胞跨膜肽R9与人铜、锌超氧化物歧化酶(superoxide dismutase 1,SOD1)和谷胱甘肽S-转移酶(glutathione S-transferase,GST)之间融合 MMP-2/9的底物肽X,设计了融合蛋白GST-SOD1-X-R9.该蛋白在肿瘤微环境中可因MMP-2/9酶切底物肽X而失去跨膜肽,从而无法进入肿瘤细胞,进而只能进入正常细胞.全基因合成SOD1-X-R9序列,并将其插入原核表达载体pGEX-4T-1中,得到表达质粒,并实现了GST-SOD1-X-R9融合蛋白的可溶表达.GST-SOD1-X-R9经硫酸铵沉淀和GST亲和层析纯化,分子量约为47kDa,与理论值一致.纯化的融合蛋白的SOD活性和GST活性分别为2 954 U/mg和328 U/mgoGST-SOD1-X-R9的SOD活性或GST活性在生理条件下几乎没有变化.该融合蛋白在溶液中可被胶原酶Ⅳ部分水解.分别建立了2D和3D培养的HepG2细胞模型来检验肿瘤微环境中的MMP-2活力对该蛋白跨膜能力的影响.在2D培养模型中,HepG2的MMP-2活力极低,但在3D培养模型中,随着培养时间的增加,HepG2肿瘤球的体积变大,其胞外MMP-2活力也随之增强.GST-SOD1-X-R9在2D培养的HepG2细胞中具有和GST-SOD1-R9蛋白一样的跨膜效率,但在3D培养的HepG2细胞球中的跨膜能力大大降低.本研究为后续深入研究GST-SOD1-X-R9靶向防护正常细胞的氧化损伤效应奠定了基础.

Keyword :

MMP-2/9敏感性 MMP-2/9敏感性 可跨膜 可跨膜 大肠杆菌表达体系 大肠杆菌表达体系 抗氧化酶 抗氧化酶 稳定性 稳定性 纯化 纯化

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GB/T 7714 何火聪 , 林丽香 , 李玲玲 et al. 基质金属蛋白酶-2/9敏感型可跨膜融合抗氧化酶的表达、纯化和表征 [J]. | 生物工程学报 , 2022 , 38 (9) : 3515-3527 .
MLA 何火聪 et al. "基质金属蛋白酶-2/9敏感型可跨膜融合抗氧化酶的表达、纯化和表征" . | 生物工程学报 38 . 9 (2022) : 3515-3527 .
APA 何火聪 , 林丽香 , 李玲玲 , 吴伦巧 , 林海英 , 潘剑茹 . 基质金属蛋白酶-2/9敏感型可跨膜融合抗氧化酶的表达、纯化和表征 . | 生物工程学报 , 2022 , 38 (9) , 3515-3527 .
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基质金属蛋白酶-2/9敏感型可跨膜融合抗氧化酶的表达、纯化和表征(英文) CSCD PKU
期刊论文 | 2022 , 38 (09) , 3515-3527 | 生物工程学报
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Abstract :

融合了跨膜肽的抗氧化酶可进入细胞,保护细胞免受放射损伤。然而跨膜肽的跨膜能力没有靶向性,其也可把抗氧化酶带入肿瘤细胞进而保护肿瘤细胞,降低放疗的效果。为此,根据多数肿瘤细胞微环境中存在活性基质金属蛋白酶(matrix metalloproteinase,MMP)-2或MMP-9的特点,在细胞跨膜肽R9与人铜、锌超氧化物歧化酶(superoxide dismutase 1,SOD1)和谷胱甘肽S-转移酶(glutathione S-transferase,GST)之间融合MMP-2/9的底物肽X,设计了融合蛋白GST-SOD1-X-R9。该蛋白在肿瘤微环境中可因MMP-2/9酶切底物肽X而失去跨...

Keyword :

MMP-2/9敏感性 MMP-2/9敏感性 可跨膜 可跨膜 大肠杆菌表达体系 大肠杆菌表达体系 抗氧化酶 抗氧化酶 稳定性 稳定性 纯化 纯化

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GB/T 7714 何火聪 , 林丽香 , 李玲玲 et al. 基质金属蛋白酶-2/9敏感型可跨膜融合抗氧化酶的表达、纯化和表征(英文) [J]. | 生物工程学报 , 2022 , 38 (09) : 3515-3527 .
MLA 何火聪 et al. "基质金属蛋白酶-2/9敏感型可跨膜融合抗氧化酶的表达、纯化和表征(英文)" . | 生物工程学报 38 . 09 (2022) : 3515-3527 .
APA 何火聪 , 林丽香 , 李玲玲 , 吴伦巧 , 林海英 , 潘剑茹 . 基质金属蛋白酶-2/9敏感型可跨膜融合抗氧化酶的表达、纯化和表征(英文) . | 生物工程学报 , 2022 , 38 (09) , 3515-3527 .
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Expression, purification and characterization of the full-length SmpB protein from Mycobacterium tuberculosis SCIE
期刊论文 | 2018 , 151 , 9-17 | PROTEIN EXPRESSION AND PURIFICATION
WoS CC Cited Count: 3
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The trans-translation system is recognized as an excellent target for developing new drugs to rapidly sterilize Mycobacterium tuberculosis (TB) infection and significantly shorten TB treatment duration. As a vital component of the trans-translation system for rescuing stalled ribosomes, the SmpB protein from Mycobacterium tuberculosis (MtbSmpB, 1-160 a. a.) mediates tmRNA binding to stalled ribosomes through forming a complex with tmRNA. So far, few works have been conducted to prepare, characterize biophysical properties and determine three-dimensional structure for the full-length MtbSmpB protein. In the present work, we successfully expressed and purified the His-tagged full-length MtbSmpB protein in Escherichia coli with a yield of 26.9 mg from 1 L of Luria Bertani medium. We also obtained MtbSmpB with a yield of 18.5 mg from 1 L of M9 minimal medium. The MtbSmpB protein showed a single band in SDS-PAGE with a molecular weight of similar to 20 kDa consistent with the measurement from MALDI-TOF-mass spectrometry. The dynamic light scattering experiment indicated that MtbSmpB existed in a monomeric form. Moreover, both circular dichroism and nuclear magnetic resonance (NMR) experiments exhibited that MtbSmpB was well structured, suggesting that it could be feasible to determine its solution structure by NMR spectroscopy. NMR titration experiments showed that MtbSmpB specifically bound to tmRNA. This work lays the essential basis for further determining the solution structure and dynamics of the full-length MtbSmpB protein.

Keyword :

MtbSmpB MtbSmpB NMR NMR Protein expression and purification Protein expression and purification Protein-tmRNA interaction Protein-tmRNA interaction Tuberculosis Tuberculosis

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GB/T 7714 Yang, Juanjuan , Liu, Yindi , Xu, Shuli et al. Expression, purification and characterization of the full-length SmpB protein from Mycobacterium tuberculosis [J]. | PROTEIN EXPRESSION AND PURIFICATION , 2018 , 151 : 9-17 .
MLA Yang, Juanjuan et al. "Expression, purification and characterization of the full-length SmpB protein from Mycobacterium tuberculosis" . | PROTEIN EXPRESSION AND PURIFICATION 151 (2018) : 9-17 .
APA Yang, Juanjuan , Liu, Yindi , Xu, Shuli , Lin, Haiying , Meng, Chun , Lin, Donghai . Expression, purification and characterization of the full-length SmpB protein from Mycobacterium tuberculosis . | PROTEIN EXPRESSION AND PURIFICATION , 2018 , 151 , 9-17 .
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肿瘤特异性嵌合抗原受体EGFRvⅢ-CAR的构建及体外活性分析 CSCD PKU
期刊论文 | 2018 , 34 (2) , 221-228 | 中国生物化学与分子生物学报
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Abstract :

嵌合抗原受体T细胞免疫疗法(chimeric antigen receptor-T,CAR-T),是通过体外激活和扩增肿瘤特异或非特异性杀伤细胞达到抗肿瘤效果,在肿瘤免疫治疗方面具有良好的应用前景.本研究构建靶向EGFRⅢ(epidermal growth factor receptor variant Ⅲ)的嵌合抗原受体(CAR)的重组慢病毒表达载体,利用慢病毒感染并筛选能够稳定表达该嵌合抗原受体的Jurkat细胞系.通过EGFRvⅢ分子刺激、与U87MG细胞共培养的方式检测细胞系的活化状况.结果显示,成功构建了pCDH-EGFRvⅢscFv-CAR-copGFP-T2A-puro慢病毒表达重组质粒,并筛选出可稳定表达EGFRⅢ-CAR的Jurkat细胞系.CCK-8法检测显示,EGFRvⅢ分子刺激12h的Jurkat-CAR细胞增殖率约是对照组的1.36倍(P<0.05);ELISA法检测显示,与U87MG细胞共孵育后,细胞上清中IL-2的浓度约是单独培养分泌在上清中IL-2的1.625倍(P<0.01).以上结果表明,稳定表达CAR的iurkat细胞,可以靶向性识别EGFRvⅢ分子及EGFRvⅢ阳性的靶细胞,并引起IL-2细胞因子释放,为后续临床细胞免疫治疗提供了理论基础.

Keyword :

EGFRvⅢ EGFRvⅢ Jurkat细胞 Jurkat细胞 嵌合抗原受体 嵌合抗原受体 慢病毒 慢病毒

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GB/T 7714 高蕊 , 詹杜清 , 杜明明 et al. 肿瘤特异性嵌合抗原受体EGFRvⅢ-CAR的构建及体外活性分析 [J]. | 中国生物化学与分子生物学报 , 2018 , 34 (2) : 221-228 .
MLA 高蕊 et al. "肿瘤特异性嵌合抗原受体EGFRvⅢ-CAR的构建及体外活性分析" . | 中国生物化学与分子生物学报 34 . 2 (2018) : 221-228 .
APA 高蕊 , 詹杜清 , 杜明明 , 张芸 , 董欣 , 祝婧雯 et al. 肿瘤特异性嵌合抗原受体EGFRvⅢ-CAR的构建及体外活性分析 . | 中国生物化学与分子生物学报 , 2018 , 34 (2) , 221-228 .
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肺炎链球菌表面蛋白A的家族Ⅰ不同支系融合蛋白的制备及免疫原性研究 CSCD PKU
期刊论文 | 2015 , 35 (05) , 377-381 | 中华微生物学和免疫学杂志
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Abstract :

目的构建肺炎链球菌家族Ⅰ中支系1和2肺炎链球菌表面蛋白A(PspA)重组融合蛋白,分析融合蛋白的免疫原性。方法 PCR扩增目的基因片段,构建重组质粒并表达,ELISA检测抗体滴度和亲和度,以及抗蛋白血清特异性和免疫交叉性。结果成功构建重组质粒并原核表达。血清中抗体滴度可达到1×10~4,亲和度达到2×10~5,调理吞噬试验阳性,保护试验效果显著。结论肺炎链球菌家族Ⅰ中支系1和2肺炎链球菌表面蛋白A(PspA)重组融合蛋白具有较高的免疫原性,能够诱导小鼠产生高滴度和高特异性的抗体,具有免疫交叉保护性。

Keyword :

免疫原性 免疫原性 肺炎链球菌 肺炎链球菌 肺炎链球菌表面蛋白A 肺炎链球菌表面蛋白A 重组蛋白 重组蛋白

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GB/T 7714 林海英 , 彭永辉 , 张双玲 et al. 肺炎链球菌表面蛋白A的家族Ⅰ不同支系融合蛋白的制备及免疫原性研究 [J]. | 中华微生物学和免疫学杂志 , 2015 , 35 (05) : 377-381 .
MLA 林海英 et al. "肺炎链球菌表面蛋白A的家族Ⅰ不同支系融合蛋白的制备及免疫原性研究" . | 中华微生物学和免疫学杂志 35 . 05 (2015) : 377-381 .
APA 林海英 , 彭永辉 , 张双玲 , 罗春华 , 郑美云 , 吕唯 et al. 肺炎链球菌表面蛋白A的家族Ⅰ不同支系融合蛋白的制备及免疫原性研究 . | 中华微生物学和免疫学杂志 , 2015 , 35 (05) , 377-381 .
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