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入门定价/纯化蛋白,1mg/ProteinD-1
商品详细scarabgenomics/非乙酰化蛋白D

入门定价/纯化蛋白,1mg/ProteinD-1
scarabgenomics/非乙酰化蛋白D入门定价/纯化蛋白,1mg/ProteinD-1
scarabgenomics/非乙酰化蛋白D

入门定价/纯化蛋白,1mg/ProteinD-1
商品编号: ProteinD-1
品牌: scarabgenomics
市场价: ¥0.00
美元价: 0.00
产地: 美国(厂家直采)
公司:
产品分类: 多肽合成
公司分类: peptide
联系Q Q: 3392242852
电话号码: 4000-520-616
电子邮箱: info@ebiomall.com
商品介绍

Background

Protein D is a 42 kDa surface-exposed lipoprotein from Haemophilus influenzae that is conserved among H. influenzae strains. It is highly immunogenic and its ability to stimulate T-cell immunity with a low risk of interference from simultaneously administered vaccines makes it a strong carrier protein candidate. The 10-valent pneumonoccal conjugate vaccine PHiD-CV licensed in 2008 (Synflorix, GSK Vaccines) uses protein D in 8 out of 10 pneumonoccal serotypes.

Scarab Genomics’ Protein D is a recombinant form expressed in Clean Genome® E. coli, our proprietary platform.

SOURCEClean Genome® E. coli expressed recombinant Protein D

PRODUCT MOLECULAR MASS:42 kiloDaltons

Figures

Scarab Genomics’ Protein D is exceptionally pure. Five replicates of Scarab’s Protein D were run on a Bis-Tris 4-12% gradient SDS polyacrylamide gel and stained with GelCode™ Blue, then purity assessed via gel scan. Average purity was 96.5%.

The sequence of Scarab Protein D was confirmed by amino acid sequencing and mass spectrometry (MS).Bioinformatics analysis of MS material revealed no post-translational modifications. Amino acid sequencing identified pure Protein D and found no norleucine incorporation. MS coverage was 92% (blue letters).

Protein D was stable for one month in storage buffer at -20oC (day 0 in each case), 4oC and 23oC. By 30 days, a slight decrease in Protein D band intensity was evident after incubation at 37oC.

Scarab’s Protein D is conjugation efficient Protein D lot 1 (PDlot1) and PDlot2 were almost as efficient in lysine conjugation as the carrier protein CRM197 and the control, BSA. Using Solulink’s ChromaLink™ Biotin conjugation assay, which uses a NHS ester reaction scheme to conjugate Biotin to primary amines (absorbance at 354 nm), we found at least 5 sites on Scarab’s Protein D that were accessible for conjugation. Far more of the 36 lysines in Protein D are likely accessible for conjugation but excessive attachment of ChromaLink Biotin causes conjugated proteins to be insoluble.

Specifications

Quality Control

Purity:
    ≥95% Protein D by SDS-PAGE
Concentration:1 mg/mL in 25mM triethanolamine, 150mM NaCl, pH7.4 via absorbance at 280 nm using an E0.1%=1.58 for a 1 mg/mL solution
Endotoxin:35 EU/mg of protein by the kinetic turbidimetric LAL method (maximum sensitivity = 0.01 EU/mL)

Grade: Protein D is for Research Use Only, Not for use in humans or as a diagnostic agent.

Storage Conditions:

  • Protein D is a 0.2 µm-filtered frozen solution of 1 mg/mL Protein D, 150mM NaCl, 25mM triethanolamine, pH 7.4. Store at -70°C upon receipt.
  • Repeated freeze-thaw cycles can degrade material. To minimize freeze-thaw cycles, aliquot Protein D to match desired use.
  • Handle product gently, DO NOT VORTEX.

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Patents & Disclaimers

Scarab is providing you with this Material subject to the non-transferable right to use the subject amount of the Material for your research at your academic institution. The Recipient agrees not to sell or otherwise transfer this Material, or anything derived or produced from the Material to a third party. NO RIGHTS ARE PROVIDED TO USE THE MATERIAL OR ANYTHING DERIVED OR PRODUCED FROM THE MATERIAL FOR COMMERCIAL PURPOSES. If the Recipient makes any changes to the chromosome of the Material that results in an invention in breach of this limited license, then Scarab will have a worldwide, exclusive, royalty-free license to such invention whether patentable or not. If the Recipient is not willing to accept the terms of this limited license, Scarab is willing to accept return of this product with a full refund, minus shipping and handling costs. For information on obtaining a license to this Material for purposes other than research, please contact Scarab’s Licensing Department. Scarab Genomics’ technology is covered by the following Patent Applications: US, 15/122,891, PCT/US2016/25588 and their related foreign applications.Clean Genome® is a registered trademark of Scarab Genomics, LLC.

品牌介绍
scarabgenomics在pDNA生产中遇到的一个常见挑战是茎环DNA结构的存在,例如病毒长末端重复序列(LTR)或短发夹RNA(shRNA),它们难以复制并且即使在特意设计的E中也使载体不稳定大肠杆菌宿主,例如Stbl3™。清洁Genome® 大肠杆菌可显着提高不稳定pDNA载体的产量。CleanGenome®E . coli Out表现出用于慢病毒生产的Stbl3™。Chakiath&Esposito(2007)研究表明,“包含慢重复序列的慢病毒表达克隆通常在大肠杆菌中表现出极大的不稳定性。。” 即使是专门设计用于克隆慢病毒直接重复序列的宿主(例如Stbl3™)也被证明是不足够的。Chakiath&Esposito表明,CleanGenome® 大肠杆菌菌株MDS™42是基因组平台减少的基础菌株,可稳定包含这些重复序列的慢病毒表达克隆(图1)。在使用MDS™42进行的100多次克隆反应中,这些作者仅选择了两个转化子菌落进行分析,在超过95%的情况下,两个克隆都具有正确的限制性图谱,从而节省了重组质粒制备的大量时间和精力。同样,衍生自腺相关病毒(AAV)的载体带有两个反向重复序列,形成40 bp茎。在末端,另外两个9bp的茎分支进一步终止于环结构。这两个茎环本身包含直接重复序列,当在标准大肠杆菌宿主中生长时,特别容易被删除。极端的例子是pT-ITR-IL2载体(图2),其中包含ITR1和ITR2茎环,它们也是彼此直接重复的。为了测试CleanGenome®菌株解决不稳定的生物治疗性pDNA生产挑战的能力,圣甲虫基因组公司使用了腺相关病毒pT-ITR-IL2载体。将其转化到MDS™42和未还原的亲本菌株大肠杆菌中K-12 MG1655。选择含有对NotI,KpnI和MscI具有正确限制模式的质粒的克隆。为了测试在大肠杆菌中的生长过程中的稳定性,将来自两个宿主的一式三份的克隆在Luria Broth中在37°C下生长24小时,并进行选择。取出培养样品进行分析,然后将每组的一种正确培养物通过在新鲜肉汤中稀释10-6 – 10-7来开始连续传代。再进行四次相似的连续传代,从每个阶段分离质粒DNA以进行分析。在MDS™42中生长的pT-ITR-IL2的限制性酶切消化模式没有随传代而改变,而MG1655生长的质粒不稳定(图3)。)。KpnI消化物(单个位点)显示出较小的质粒片段,与通过内部重组使质粒重复序列之间的“锤头”区域丢失相一致。MscI消化(每个ITR两侧的位点)同样显示出片段之一的逐渐丢失,与KpnI消化一致。DNA测序证实了这些结果。