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Enhanced potency of an IgM-like nanobody targeting conserved epitope in SARS-CoV-2 spike N-terminal domain

Signal Transduction and Targeted Therapy 2024年第6期9卷 2726-2737页

作者： Bo Liu Honghui Liu Pu Han Xiaoyun Wang Chunmei Wang Xinxin Yan Wenwen Lei Ke Xu Jianjie Zhou Jianxun Qi ruiwen fan Guizhen Wu Wen-xia Tian George F.Gao Qihui Wang College of Veterinary Medicine Shanxi Agricultural University030801 JinzhongChina CAS Key Laboratory of Pathogen Microbiology and Immunology Institute of MicrobiologyChinese Academy of Sciences(CAS)100101 BeijingChina School of Life Sciences Yunnan University650504 KunmingYunnan ProvinceChina NHC Key Laboratory of Biosafety National Institute for Viral Disease Control and PreventionChinese Center for Disease Control and Prevention(China CDC)102206 BeijingChina Savaid Medical School University of Chinese Academy of Sciences101408 BeijingChina

Almost all the neutralizing antibodies targeting the receptor-binding domain(RBD)of spike(S)protein show weakened or lost efficacy against severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)emerged or emerging variants,such as Omicron and its *** suggests that highly conserved epitopes are crucial for the development of neutralizing ***,we present one nanobody,N235,displaying broad neutralization against the SARS-CoV-2 prototype and multiple variants,including the newly emerged Omicron and its ***-electron microscopy demonstrates N235 binds a novel,conserved,cryptic epitope in the N-terminal domain(NTD)of the S protein,which interferes with the RBD in the neighboring S *** neutralization mechanism interpreted via flow cytometry and Western blot shows that N235 appears to induce the S1 subunit shedding from the trimeric S ***,a nano-IgM construct(MN235),engineered by fusing N235 with the human IgM Fc region,displays prevention via inducing S1 shedding and cross-linking virus *** to N235,MN235 exhibits varied enhancement in neutralization against pseudotyped and authentic viruses in *** intranasal administration of MN235 in low doses can effectively prevent the infection of Omicron sub-variant BA.1 and XBB in vivo,suggesting that it can be developed as a promising prophylactic antibody to cope with the ongoing and future infection.

关键词： doses prevention conserved

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痘苗病毒蛋白D8纳米抗体的原核表达及结合活性分析

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中国兽医学报 2024年第8期44卷 1728-1734页

作者：田婧婧杨晓岚李涛王慧范瑞文山西农业大学动物医学学院山西太谷030801 军事科学院军事医学研究院微生物流行病研究所病原微生物生物安全国家重点实验室北京100071

利用SUMO标签制备痘苗病毒(vaccinia virus,VACV)蛋白D8的纳米抗体,并分析其与抗原是否具有结合活性。设计并合成序列,构建原核表达质粒pKMD-SUMO-D8,将其转化大肠杆菌(Escherichia coli,***)BL21(DE3)感受态细胞并诱导表达;纯化带有Hi... 详细信息

利用SUMO标签制备痘苗病毒(vaccinia virus,VACV)蛋白D8的纳米抗体,并分析其与抗原是否具有结合活性。设计并合成序列,构建原核表达质粒pKMD-SUMO-D8,将其转化大肠杆菌(Escherichia coli,***)BL21(DE3)感受态细胞并诱导表达;纯化带有His标签和SUMO标签的抗体Anti-SUMO-D8,之后利用SUMO蛋白酶切除His标签和SUMO标签,进一步获得Anti-D8;最后,应用间接酶联免疫吸附试验(ELISA)及间接免疫荧光试验(IFA)对其结合活性进行检测并分析。结果显示,成功构建了原核表达质粒pKMD-SUMO-D8和重组菌株,在30℃条件下,终浓度为0.1 mmol/L的IPTG诱导表达菌株4 h时,Anti-SUMO-D8表达量最佳;获得了高纯度的Anti-SUMO-D8和Anti-D8纳米抗体;间接ELISA结果显示,Anti-SUMO-D8和Anti-D8与抗原均具有结合活性;在相同条件下,Anti-D8与VACV及E8L的结合活性高于Anti-SUMO-D8。间接IFA结果显示,VACV侵染48 h后,试验组部分细胞核在荧光显微镜下可见绿色荧光。利用SUMO标签可高效制备VACV蛋白D8的纳米抗体,且制备的纳米抗体具有一定的结合活性。本研究结果为纳米抗体的制备提供了新思路,也为深入研究VACV蛋白D8纳米抗体的生物学功能奠定了基础。

关键词：痘苗病毒纳米抗体 SUMO 结合活性

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Identification of a novel microRNA function in alpaca skin

Identification of a novel microRNA function in alpaca skin

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中国畜牧兽医学会动物解剖学及组织胚胎学分会第十七次学术研讨会

作者： ruiwen fan Junming Bai Shanshan Yang Changsheng Dong College of Animal Science and Veterinary Medicine Shanxi Agricultural University

From the deep sequencing of alpaca skins with white and black coat color,we obtained 15 and 18 potentially novel miRNAs from the WA and BA skin libraries,*** those,a novel miRNA expressed differently in white and black alpaca skin(named miRNA-066).According to the target gene prediction,one of the target genes is melanophilin(MLPH),which plays important role in genesis in *** get the function of miRNA-066 in alpaca skin,miRNA was over expressed in melanocyte in *** expression level of miRNA-066 was tested by qRT-PCR and results showed it expressed *** expression level of MLPH,TYR,TYRPl,TRY and KIT protein were tested by western blotting,and the results showed that TYR,TYRPl and KIT protein were *** those results suggested that miRNA-066 might regulate melanogenesis.

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Ribosomal proteins expression and phylogeny in alpaca (<i>Lama pacos</i>) skin

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Advances in Bioscience and Biotechnology 2012年第3期3卷 231-237页

作者： Junming Bai ruiwen fan Shanshan Yang Yuankai Ji Jianshan Xie Chang-Sheng Dong College of Animal Science and Veterinary Medicine Shanxi Agricultural University Taigu China

Ribosomal proteins (RP) has been reported as a central player in the translation system, and are required for the growth and maintenance of all cell kinds. RP genes form a family of homologous proteins that express in the stable pattern and were used for phylogenetic analysis. Here we constructed a cDNA library of alpaca skin and 13,800 clones were sequenced. In the cDNA library, RP genes from skin cDNA library of alpaca were identified. Then 8 RP genes were selected at random and built the phylogenetic trees from the DNA sequences by using parsimony or maximum likelihood methods for molecular and evolutionary analysis of ribosomal proteins. The results showed that the 42 RP genes of alpaca have been expressed in alpaca skin. They were highly conserved. The phylogeny inferred from all these methods suggested that the evolutionary distances of alpaca RP genes were closer to rat.

关键词： Ribosomal Protein Expression Phylogenetic Tree Alpaca

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Alpaca Management in North of China

Alpaca Management in North of China

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提升骆驼产业科技含量，打造骆驼行业知名品牌——第二届（2013）中国骆驼大会

作者： Shanshan Yang Zhanquan Shi Kaiyuan Ji ruiwen fan Changsheng Dong College of Animal Science and Technology Shanxi Agricultural University

Alpaca originates from South America and is the most important source of fine *** have been successfully introduced into many countries including Australia,Japan,New Zealand,British,and SO ***,23 huacaya alpacas had been successfully imported into China from Australia in May,2002, following the agreement signed between Australia and *** more than 3 years of living in China, alpacas reproduced more than one generation without any diseases and disorders.

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The microsatellite markers with Genetic Diversity of Alpaca(Lama pacos)

The microsatellite markers with Genetic Diversity of Alpaca(...

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中国畜牧兽医学会动物解剖及组织胚胎学分会第十九次学术研讨会

作者： Yucong Cui Jianjun Geng Qingling Geng Zengfeng Ding ruiwen fan Changsheng Dong College of Animal Science and Veterinary Medicine Shanxi Agricultural University

The polymorphisms of 17 microsatellite loci of 61 Alpaca(Lama pacos) was detected using polymerase chain reaction(PCR) and DNA sequencing technology,and the paternity testing was studied combined with the parents of this *** purpose is to supply effective microsatellite locus for germplasm resourses evaluation and paternity testing of *** demonstrated that 17 microsatellite loci altogether discovered 99 *** number of alleles was most in VOLP77(11),and the least in VOLP12(2).The heterozygosity was highest in VOLP77(0.852),while the lowest was VOLP12(0.048).The other 15 microsatellite loci were highly polymorphic except VOLP12 and VOLP01 which are inferior *** mean observed heterozygosity(Ho),the expected heterozygosity(He),and the polymorphism information content(PIC) were 0.594,0.554,and 0.454,*** above results suggested mat there were a relatively high genetic diversity in Alpaca from *** could improve the accuracy requirements of genetic diversity,make the operation more convenient,reduce costs,and be more suitable for practical production.

关键词： Microsatellite Genetic diversity Alpaca

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Ribosomal Proteins expression and phylogeny in Alpaca(Lama pacos) skin

Ribosomal Proteins expression and phylogeny in Alpaca(Lama p...

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中国畜牧兽医学会动物解剖学及组织胚胎学分会第十七次学术研讨会

作者： Bai Junming fan ruiwen Geng Jianjun Yang Shanshan Xie Jianshan Dong Chang-sheng college of Animal Science And Veterinary Medicine Shanxi Agricultural University

Ribosomal proteins(RP) has been reported as a central player in the translation system,and are required for the growth and maintenance of all cell *** genes form a family of homologous proteins that express in the stable pattern and were used for phylogenetic *** we constructed a cDNA library of alpaca skin and 13,800 clones were *** the cDNA library,RP genes from skin cDNA library of alpaca were *** 8 RP genes were selected at random and built the Phylogenetic trees from the DNA sequences by using parsimony or maximum likelihood methods for molecular and evolutionary analysis of ribosomal *** results showed that the 42 RP genes of alpaca have been expressed in alpaca *** were highly *** phylogeny inferred from all these methods suggested that the evolutionary distances of alpaca RP genes were closer to rat.

关键词： Ribosomal protein expression Phylogenetic tree alpaca

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The effects of IGF1 on the melanogenesis in alpaca melanocytes in vitro

The effects of IGF1 on the melanogenesis in alpaca melanocyt...

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中国畜牧兽医学会动物解剖及组织胚胎学分会第十九次学术研讨会

作者： Shuaipeng Hu Yu Liu Shanshan Yang Kaiyuan Ji Xuexian Liu Junzhen Zhang ruiwen fan Changsheng Dong College of Animal Science and Veterinary Medicine Shanxi Agricultural University

In order to investigate the effects of the insulin-like growth factor 1(IGF-1) on alpaca melanocyte in vitro,different dosees of IGF1(0ng/ml,10ng/ml,20ng/ml,40ng/ml) were added in the medium of alpaca *** RTCA machine was used to monitor the proliferation,quantitative real-time PCR and western blot to test the relative gene expression,ELISA to test cAMP production,and spectrum method to test the melanin *** results showed that compared to the normal melanocyte,the proliferation of melanocytes was increased within 60 h following adding *** also showed that cAMP content produced by melanocytes was increased,Microphthalmia-associtated transcription factor(MITF),Tyrosinase(TYR) and Tyrosinase Related protein 2(TYRP2) expression was increased,and melanin production with most obvious change in 10ng/ml supplementary group,when compared with the control *** results suggested that IGFl with the dose of 10 ng/ml had the important effects on the melanogenesis in alpaca melanocyte by the cAMP pathway.

关键词： IGF1 melanogenesis melanocyte alpaca

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The expression of KRT2 and its effect on melanogenesis in alpaca skins

The expression of KRT2 and its effect on melanogenesis in al...

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中国畜牧兽医学会动物解剖及组织胚胎学分会第十九次学术研讨会

作者： Yucong Cui Yajun Song Qingling Geng Zengfeng Ding Yilong Qin ruiwen fan Changsheng Dong Jianjun Geng College of Animal Science and Veterinary Medicine Shanxi Agricultural University Tianshui Entry-Exit Inspection and Quarantine Bureau

In order to investigate the effects of the keratin 2(KRT2) on alpaca melanocyte in vivo and vitro,the immunohistochemistry(IHC),quantitative real-time PCR(qPCR),Western blot,and alpaca melanocytes transfection methods were *** results showed that mRNA and protein expression of KRT2 was highly expressed in brown skin in comparison with that in white ***,we found that KRT2 was expressed in alpaca melanocytes in vitro by *** transfection with KRT2 in alpaca melanocytes,the relative mRNA and protein expression of KRT2,microphthalmia-associtated transcription factor(MITF),tyrosinase(TYR) and tyrosinase-related protein 1(TYRP1) in alpaca skin melanocytes was increased with significant differences;a further result was the increase of melanin *** results suggested that KRT2 functions in alpaca hair color formation,which offered an essential theoretical basis for further exploration of the role of melanogenesis.

关键词： KRT2 Hair color Alpaca Melanocyte Skin

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Identification of a novel miRNA important for melanogenesis in alpaca(Lama Pacos)

Identification of a novel miRNA important for melanogenesis ...

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提升骆驼产业科技含量，打造骆驼行业知名品牌——第二届（2013）中国骆驼大会

作者： ruiwen fan Shanshan Yang Zhanquan Shi Kaiyuan Ji Xiaoyun Jia Jianbo Yao George W Smith Changsheng Dong

The mechanism of coat color formation is complex,microRNAs play important roles in the control of melanogenesis and coat color in *** the previous deep sequencing of alpaca skins,one of novel miRNAs expressed differently in white and black alpaca skin(named alp-miR066).According to the target gene prediction,one of the target genes is soluble guanylate cyclase(sGC).To get the function of alp-miR-066 in alpaca skin,it was over expressed in alpaca melanocytes in *** expression of miRNA-066 was tested by real-time PCR and the results showed it expressed *** the pathway of cAMP in melanocytes,cGMP was increased by ELASA assay,while cAMP was decreased. The expression of sGC,TYR,TYRP1,TYRP2 and MITF protein were tested by western blotting,and the results showed that sGC was increased and TYR,TYRP1 and MITF protein were ***,the number of melanogenic melanocytes was less and the amount of melanins production was 0.7 fold in alp-miR066 transfection group,compared to negative control(NC) transfection *** those results suggested that alp-miR-066 might regulate melanogenesis on the pathway of cAMP in alpaca melanocytes by targeting sGC.

关键词： alp-miR066 melanocytes cAMP melanin sGC

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