人源性抗HBsAg Fab抗体的发酵生产研究

为了适应工业生产的需要,利用fed—batch方法,重组人源性抗HBsAg Fab抗体酵母工程菌在30L发酵罐中进行了高密度发酵,发酵最适温度30℃,pH值范围5．0～5．3,溶氧范围20％～30％。发酵液OD600值达到300时开始诱导,甲醇最佳诱导浓度为10mL／L。重组人源性抗HBsAg Fab抗体经离子交换层析纯化,纯化产品经SDS-PAGE、Western blot进行分析和ELISA方法进行活性测定。结果显示,重组Fab抗体在Fed-batch发酵系统中可高效表达,经过192h的发酵生产,重组人源性抗HBsAg Fab抗体的表达量可达412mg／L。发酵上清经过离子交换层析纯化,获得纯度为95％的重组Fab抗体,该Fab抗体经ELISA分析具有较高的HBsAg抗原亲和力和特异性。结果证实可以通过高密度发酵毕赤酵母工程菌来高效生产重组人源性抗HBsAg Fab抗体,为后续的工业化生产应用奠定了基础。     

A Stable Upstream Stem-loop Structure Enhances Selection of the First 5'-ORF-AUG as a Main Start Codon for Translation Initiation of Human ACAT1 mRNA

Acyl-coenzyme A:cholesterol acyltransferase (ACAT)is an integral membrane protein, which is mainly locatedin rough endoplasmic reticulum (ER), and is responsiblefor catalyzing the intracellular formation of cholesterylester from cholesterol and long-chain fatty acyl-coenzymeA [1,2]. Human ACAT1 cDNA K1 was firstly cloned andfunctionally expressed in 1993 [3]. Further studies withspecific anti-ACAT1 antibody (DM10) illustrated that onemajor 50 kD ACAT1 protein was expressed in various…     

Phylogenetic clusters of rhizobia revealed by genome structures

Rhizobiaareagriculturallyandenvironmentallyimportantbacteria.Theirsymbiosiswithleguminousplantsisresponsibleformostoftheatmosphericni-trogenfixedonland.Classificationofthesebacteriabasedontheirnaturalrelationshipswillpromotetheirapplication.Thisresearchemploysanewphylogeneticmethod,i.e.,revelationandcomparisonofgenomestructure,tocategorizerhizobia.Phylogenyisthestudyoftheevolutionaryrelationshipsamongorgan-isms[1].Currentlyphylogeneticrelationshipsamongrhizobiaaremostlyinferredfromcomparisons…     

Differentiation potential of bone marrow mesenchymal stem cells into retina in normal and laser-injured rat eye

Withinthebonemarrowstromathereexistsasubsetofnonhematopoieticcellsreferredtoasmes-enchymalstemormesenchymalprogenitorcells.Mesenchymalstemcells(MSCs)areapopulationofpluripotentcellswithinthehuman,birdorrodentbonemarrowmicroenvironmentdefinedbytheirability,eitherinvitroorinvivo,todifferentiateintocellsoftheosteogenic,chondrogenic,tendonogenic,adipo-genic,neuralcellsandmyogeniclineages[1].Themethodologiestoisolateandculture-expandMSCsfromhumanbonemarrowforestablishingthecellularortissuediffere…     

Three-dimensional structure determination of capsid of Aedes albopictus C6/36 cell densovirus

Apreviously-unknownviruswasfoundchroni-callyinfectingC6/36cellsthatwereusedtoculturedenguevirus.Afterisolationandpurification,RT,PCRandsequencingwereperformedwithrandomprimers.TheresultsshowthattheviralgenomeisssDNAandis4096ntinlength(GenBankAccessionNo.AY095351).Thenucleicacidsequenceofthispreviously-undescribedvirussharesabout90%iden-titywiththatofAedesaegyptidensonucleosisvirusandtheaminoacidsequencesofthreeproteinsen-codedbythenucleicacidshareabout89%—93%identitieswiththoseofAedesa…     

Developmental patterns of serum leptin levels, leptin gene expression in adipose tissue and Ob-Rb gene expression in hypothalamus of Erhualian and Large White pigs

~~Developmental patterns of serum leptin levels, leptin gene expression in adipose tissue and Ob-Rb gene expression in hypothalamus of Erhualian and Large White pigs~~     

Chromatography studies on bio-affinity of nine ligands of a1-adrenoceptor to a1D subtypes overexpressed in cell membrane

Copyright by Science in China Press 2004 The a1-AR is present in many tissues including heart, blood vessels, liver, brain, kidney, prostate and spleen. In these tissues, the a1-AR mediates a variety of physiological effects such as neurotransmission, vasoconstriction, cardiacinotropy, chronotropy and glycogenolysis[1]. In 1986, Morrow and Greese pro-posed that a1-AR should be subclassified into two subtypes. Later this hypothesis was proved by Q.D Han in 1987. He not only verified a1-A…     

The complete mitochondrial genomes sequences of Asio flammeus and Asio otus and comparative analysis

Mitochondrial DNA (mtDNA), as a model sys-tem, has been extensively used for molecular phy-logenetic and evolutionary analysis[1]. With the ad-vances in DNA sequencing technology, more andmore researchers prefer to use complete mitochondrialgenome for phylogenetic analysis[2—4]. Since the com-plete sequencing of human mtDNA in 1981 (Andersonet al., 1981)[5], 342 vertebrate mitochondrial genomeshave so far been sequenced. Up to now the completesequences of 29 avian mitochondrial genomes h…     

Screening and identification of Shigella flexneri 2a virulence-related genes induced after invasion of epithelial cells

~~Screening and identification of Shigella flexneri 2a virulence-related genes induced after invasion of epithelial cells1. Jin, Q., Yuan, Z., Xu, J., Wang, Y., Shen, Y., Lu, W., Wang, J., Liu, H., Yang, J., Yang, P., Zhang, X., Zhang, J., Yang, G, Wu, H., Qu, D., Dong, J., Sun, L., Xue, Y, Zhao, A., Gao, Y., Zhu, J., Kan, B., Ding, K.. Chen, S., Cheng, H., Yao, Z., He, B., Chen, R., Ma, D., Qiang, B., Wen, Y, Hou, Y., Yu, J., Genome sequence of Shigella flexneri 2…     

Identification and expression profiling of putative odorant-binding proteins in the malaria mosquitoes, Anopheles gambiae and A.arabiensis

~~Identification and expression profiling of putative odorant-binding proteins in the malaria mosquitoes, Anopheles gambiae and A. arabiensis1. Curtis, C. F., Introduction 1: An overview of mosquito biology, behaviour and importance, in Olfaction in Mosquito-Host Interactions (eds. Bock, G. R.. Cardew, G.), New York: Wiley, 1996, 3-7. 2. Nighom, A., Hildebrand. J. G.. Dissecting the molecular mechanisms of olfaction in a malaria-vector mosquito, PNAS, 2002, 99(3): 1113-…