蛋白表达服务

1.用于提高基因表达水平的OptimumGene^TM密码子优化技术

金斯瑞先进的OptimumGene^TM密码子优化技术能帮助您优化基因序列，使基因表达和蛋白溶解度达到较高水平。OptimumGene^TM技术还可用来优化很多与转录、翻译、蛋白折叠相关的参数。

2.较高的成功率以及丰富的重组蛋白制备经验

我们已经为客户制备了数以千记的复杂蛋白,其中包括：跨膜蛋白,分泌蛋白,蛋白酶以及其它的酶类,成功率均在90%以上。金斯瑞制备的重组蛋白已成功应用于靶点确认、酶鉴定、抗体制备、高通量筛（HTS）、 构象研究以及SAR（结构-活性关系）。

3.BacPower^TM技术和FoldArt^TM蛋白折叠技术

金斯瑞自主研发的BacPower^TM技术和FoldArt^TM蛋白折叠技术，使蛋白难表达和蛋白难溶等问题迎刃而解。

4.全面的服务

金斯瑞能为您提供基因合成，蛋白表达，稳定细胞株生产，抗体制备以及免疫检测等一体化服务。

5.高质量的服务

金斯瑞高质量的蛋白表达服务，可满足您对蛋白的纯度高、活性强、交付时间快等需求。金斯瑞交付的蛋白一般纯度在95%以上，内毒素水平低于0.01 EU/µg。

药物研发过程中,制备和纯化后的蛋白具有生物活性是十分必要的。金斯瑞先进的设备和专利的制备方法,保证了交付给客户的蛋白均具有生物活性。我们进行蛋白表达与免疫检测相结合,交付给客户的高质量、高活性蛋白,甚至超过了客户的预期,具体请参考下面的示例。我们通过对细胞破壁方法进行优化,提高了纯度大于90%的目标蛋白的获得量。

• Fig.A: SDS-PAGE analysis of the fractions upon the IEX chromatography 。
• Lane 1-47: Gradient elution fractions
• Lane M: Low weight marker (Jinsite, Cat. No. MM0906)
• Lane S: The supernatant after homogenizer with high pressure
• Lane ft: Flow through

• Fig.B: Enzyme assay monitoring the fractionation by DEAD Sepharose

• Fig. C: Fractions pooled based on activity analysis for further ADP Sepharose affinity purification
• Fig. D: SDS-PAGE analysis of the purified protein
• Lane M: Low weight marker (Jinsite, Cat. No. MM0906)
• Lane S: The supernatant after homogenizer with high pressure
• Lane ft: Flow through

金斯瑞专利的蛋白折叠技术,可以解决原核系统内由于过量表达而带来的蛋白活性丧失、折叠错误、蛋白发生集聚反应不能溶解等问题。我们可以成功溶解并折叠含有95%以上包涵体的目标蛋白（请参考下述示例）,折叠蛋白的纯度高达85%以上。金斯瑞交付给客户的折叠蛋白储存于特制缓冲液中。

• Fig. Refolded protein of sample was analyzed by SDS-PAGE followed by Coomassie Brilliant Blue staining
• Lane M: Smart Protein Standard (Jinsite, Cat. No. MM0900)
• Lane 1: Refolded protein of sample

重组蛋白中如果含有表位附加标签，可能会导致蛋白结构改变、产生毒性，甚至丧失活性。为了避免这类问题，很多客户在蛋白表达后要求去除标签。在下面的示例中，金斯瑞使用SUMO蛋白酶，成功去除了标签。获得的目标蛋白标签去除率为80%，纯度高达90%以上。

SDS-PAGE checking purification of protein after tag cleavage by SUMO protease

Fig。 A:

• Lane M: Smart Protein Standard (Jinsite, Cat.No. MM0900)
• Lane 1: Refolded fusion protein with SUMO-tag

Fig. B:

• Lane M: Smart Protein Standard (Jinsite, Cat.No. MM0900)
• Lane 1: Recovered protein after 1st incubation with Ni-resin
• Lane 2: Recovered protein after 2nd incubation with Ni-resin
• Lane 3: Elute with 300 mM imidazole from Ni-resin of 2nd incubation

• Youngjin Park, Mohd Amir F. Abdullah, Milton D. Taylor, Khalidur Rahman, and Michael J. Adang. Enhancement of Bacillus thuringiensis Cry3Aa and Cry3Bb toxicities to coleopteran larvae by a toxin-binding fragment of an insect cadherin. Appl. Envir. Microbiol. May 2009; 75(10): 3086-3092
• Ilene M. Pedroso, William Hayward, and Terace M. Fletcher. The effect of the TRF2 N-terminal and TRFH regions on telomeric G-quadruplex structures. Nucleic Acids Res. Apr 2009; 37(5): 1541-1554
• Huiyong Wei, Dan Huang, Jeff Fortman, Richard Wang, Linyun Shao, and Zheng W. Chen. Coadministration of Cidofovir and Smallpox Vaccine Reduced Vaccination Side Effects but Interfered with Vaccine-Elicited Immune Responses and Immunity to Monkeypox. J. Virol. Jan 2009; 83(2): 1115 - 1125
• Ashish Nagarsekar, Rachel S. Greenberg, Nirav G. Shah, Ishwar S. Singh, and Jeffrey D. Hasday. Febrile-Range Hyperthermia Accelerates Caspase-Dependent Apoptosis in Human Neutrophils. J. Immunol. Aug 2008; 181(4): 2636 - 2643
• Brando C, Ware LA, Freyberger H, Kathcart A, Barbosa A, Cayphas S, Demoitie MA, Mettens P, Heppner DG, Lanar DE. Murine immune responses to liver-stage antigen 1 protein FMP011, a malaria vaccine candidate, delivered with adjuvant AS01B or AS02A. Infect. Immun. Feb 2007; 75(2): 838-845
• Buchko GW, Ni S, Robinson H, Welsh EA, Pakrasi HB, Kennedy MA. Characterization of two potentially universal turn motifs that shape the repeated five-residues fold--crystal structure of a lumenal pentapeptide repeat protein from Cyanothece 51142. Protein Sci. Nov 2006; 15(11): 2579-2595
• Dieterich DC, Link AJ, Graumann J, Tirrell DA, Schuman EM. Selective identification of newly synthesized proteins in mammalian cells using bioorthogonal noncanonical amino acid tagging (BONCAT). Proc. Natl. Acad. Sci. USA. Jun 2006; 103(25): 9482-9487