ProImmune REVEAL® MHC-peptide 结合化验

寻找您需要知道的世界上引用最多次数的MHC-peptide 结合化验技术服务

体外 MHC-peptide 结合化验能够做到预测工具做不到的事。这技术化验可以确认物理结合的特性在于肽的氨基酸序列。其外,这技术化验也能够验证您开发的电脑预测工具的准确性。

ProImmune 是全球领先在于提供体外 MHC-peptide 结合化验,能够提供范围最广的 MHC 等位基因和化验选项,从高通量赛选到详细的竞争分析在于候选肽。我们专业的免疫学顾问能够协助您的需求和快速的提供最合适的化验方案。

ProImmune REVEAL® 数据从美国 FDA 的研究员发表:

美国 FDA 的研究员和同事用了 ProImmune REVEAL 了解免疫原性在于治疗的Factor VIII 蛋白质的差别在于不同的种族基于他们的 HLA 分型 (Pandey G.S. et al. PLOS Computational Biology May 2013, Vol. 9 (5) e1003066):


Figure 1: (A) Binding scores of FVIII wild type peptides. Binding scores in a REVEAL assay were measured for the synthetic wild type peptides depicted in bold in Table 1. Each row represents a peptide and each column a MHC-II allele. The peptides are grouped together based on the position in the FVIII gene where the haplotypes H2 to H6 occur. Overlapping peptides were used around the position of each ns-SNP. Positive-control (PC) peptides are known T-cell epitopes of FVIII and negative control (NC) peptides are from regions of the FVIII protein where no T-cell epitopes have been identified. The scale for the binding score is shown at the top of the figure; dark colors represent peptides that bind with high affinity to the MHC-II allele. (B) FVIII high affinity binders. Wild type peptides with binding score >15% are shown and considered potential T-cell epitopes for each of the six MHC-II alleles. (C) Binding promiscuity scores for the wild type FVIII peptides. The binding promiscuity score for each peptide, defined as the fraction of MHC-II variants the peptide binds with a score >15%, is shown.

ProImmune REVEAL® 能使用在许多病毒领域包括癌症和感染病毒。新的 CD4+ 和 CD8+T 细胞表位鉴定从这个技术的协助下可以用来取数蛋白质的免疫原性或组件在于研发疫苗或目标在于免疫治疗。


Figure 2: ProImmune REVEAL 化验在于 MHC Class I 和 II 的例子工艺流程。
  1. 在每个案例,肽都会合成在相对的项目,基于 ProImmune 用途广泛的thinkpeptides 肽合成平台。用的肽可以是在需多不同合适的构成,可以从重叠序列或单独选项的肽。
  2. 在每个案例,肽一起和重组体 MHC 分子孵化然后 MHC-peptide 复合体的稳定性是用免疫测定形式检测。
    1. 组转和离解在每个案例可以测高通量在单个时间点的间隔,在一个负担的起的价格,这能够同时赛选许多等位基因和肽。
    2. 或者是要更近在于有感兴趣的肽,更深入的速率测定可以建立一个比较详细的结合和分离率的特性。
  3. 表位鉴定可以用 MHC-multimers,比如 ProVE® Pentamers 在于 MHC Class I 或 ProT2® Tetramers 在于 MHC Class II。或者,一同组合 MHC Multimer 的科技,功能测定比如 ELISpot 化验,流式细胞术在于细胞内细胞因子染色,或增殖化验可以执行功能确认在于具体的肽。

赞美对于 ProImmune REVEAL®

Prof. Julio Delgado, University of Utah, USA “The advantage of ProImmune’s in vitro system is that we could characterize binding to each HLA molecule individually and not have to guess which of the range of HLA molecules expressed by our patients was responsible for antigen presentation. “ 

Dr. Gene Olinger, United States Army Medical Research Institute of Infectious Diseases (USAMRID) “ProImmune accomplished in two months what would have taken three years in our laboratory”.

您会得到什么?

ProImmune REVEAL® 的数据是报告在我们详细的报告型板对于这个化验。报告给的数据是在特选的化验。


Figure: REVEAL Class II 例子数据报告结合的百分点比率对阳性显示成绿色。红色的覆盖:稳定指数孵化过了24小时在37度。
Class I HLAClass II: HLA DRClass II: HLA DQ
A*01:01A1*01:01/B1*01:01A1*01:01/B1*05:01
A*02:01A1*01:01/B1*15:01A1*05:01/B1*03:01
A*03:01A1*01:01/B1*03:01A1*01:02/B1*05:02
A*11:01A1*01:01/B1*04:01A1*01:02/B1*06:02
A*24:02A1*01:01/B1*11:01A1*03:01/B1*03:02
A*29:02A1*01:01/B1*13:01A1*01:02/B1*06:04
B*07:02A1*01:01/B1*07:01A1*05:01/B1*02:01
B*08:01A1*01:01/B1*01:02A1*02:01/B1*02:02
B*14:02A1*01:01/B1*04:02A1*03:01/B1*03:01
B*15:01A1*01:01 /B1*04:04A1*02:01/B1*03:03
B*27:05A1*01:01/B1*04:05A1*03:03 /B1*03:03
B*35:01A1*01:01/B1*04:07Class II: HLA DP
B*40:01A1*01:01 /B1*04:08A1*01:03/B1*01:01
Class I: H-2A1*01:01/B1*09:01A1*01:03/B1*03:01
H-2KbA1*01:01/B1*10:01A1*01:03/B1*04:01
H-2DbA1*01:01/B1*11:02A1*01:03 /B1*04:02
H-2LdA1*01:01/B1*11:03A1*01:03/B1*05:01
H-2KdA1*01:01/B1*11:04A1*02:01/B1*01:01
H-2DdA1*01:01/B1*15:02A1*02:01/B1*02:01
Class I: MamuA1*01:01/B1*16:01A1*02:01/B1*04:01
Mamu-A*01A1*01:01/B1*16:02A1*02:01/B1*04:02
Mamu-A*02A1*01:01/B3*02:02A1*02:01/B1*05:01
Class II: IAA1*01:01/B5*01:01A1*02:01/B1*09:01
H-2IAbA1*02:01/B1*11:01
H-2IAdA1*02:01/B1*13:01

高通量的 ProImmune 数据在于 Remicade®

Figure 4: ProImmune REVEAL® 的数据显示影响到的群在于 Class II HLA 结合的肽代表一部分的小鼠变量部位在于 Remicade® (Infliximab). Remicade®,是个非常成功的药物在于治疗发炎自生免疫病毒也是显示了快速的免疫原性发生率。ProImmune REVEAL® 本生的高通量能够让研究员允许全面的了解这个单克隆抗体或治疗蛋白质和 HLA 分子互动的操作方式从一个全球的群中基于在实际的结合实验分析。

ProImmune REVEAL® 的应用

  • 快速,全面的表位图,解决 MHC 结合的实验
  • 研究免疫原性在于不同变量的蛋白质, 比如比较变量和保存的区域在于免疫原性的蛋白质在不同进化枝的病毒
  • 优化临床检测,比如选择最优良的特异性 Pentamer 做检测在一个目标的病人组
  • 鉴定交叉提成的肽在于不同的等位基因
  • 进步疫苗的设计和功效
  • 诱变研究:优化表位结合在于治疗用途,比如插入氨基酸在肽的结合位置增强亲和力

ProImmune REVEAL® 的优点

  • 物理化验。实验成绩克服了假想预测 MHC-peptide 结合
  • 快速。可得到数据在一个短时间内
  • 灵活性。ProImmune REVEAL® 提供许多选项在高通量检测从高数量的肽对一个大范围的等位基因到深入检测少数量的肽对特定 MHC 分型。我们的免疫销售专家团队能够配置一份计划书达到您的目的
  • 负担能力。ProImmune REVEAL®的可扩展性和灵活性能够设计一个项目合适您的预算目的。我们可以找出最重要的问题需要解答和设计项目能够管理程序的风险

进步知识产权地位

鉴定单独表位在于治疗蛋白质或其他有感兴趣的蛋白质可分离知识产权的蛋白质从知识产权的给药方式或治疗方式。在研发疫苗的过程,定义知识产权在于特定表位增加选项在以后的产品设计改造。如果只是产权在于蛋白质的特定抗原没有固定,竞争公司有可能开发阻塞的专利地位。

经常问起的问题

问:需要提供什么资料才能得到 ProImmune REVEAL® 项目的报价

答:理想地是我们需要得到您要赛选的肽和 MHC 等位基因。但是,如果还没有这些资料,我们可以提供指示性的报价显示肽的数量和等位基因而已,这会是一个好的起步点。

问:这个技术服务需要多久?

答:这个需要看项目的范围,包括数量和特定用的肽。当我们知道您的项目范围后我们会在报价里提供您预计的交货时间。

问:有那么多的选项,怎么选哪一个是最合适呢?

答:基于在和你们沟通的时候,了解您的需求,我们会推荐最合适的项目范围给您然后再讨论这个项目怎么能一起适合在您的研究目的和其他的化验。

案例分析:

ProImmune REVEAL® & ProVE® Rapid Epitope Discovery System used to identify novel T cell epitopes in HIV-1

Westrop, S.J. et al. (2009). “Novel approach to recognition of predicted HIV-1 Gag B35:01-restricted CD8 T-cell epitopes by HLA-B35:01+ patients: Confirmation by quantitative ELISpot analyses and characterisation using multimers.” J Immunol Methods. 341: 76-85. [PubMedID: 19056394]

Figure: Results of the MHC-peptide binding assay for controls and the 12 peptides that were considered potential epitopes; binding signal is shown as a percentage relative to the pass/fail control; peptide 8, HPVHAGPIA (red) was further validated by Pentamer staining.
Figure: Pro5® Pentamer staining of live lymphocytes gated on CD3+ cells; the left plot shows staining with an allele mismatched negative control Pentamer, the right plot shows staining with the HA9 (B*35:01/HPVHAGPIA) Pentamer. 0.39% of CD3+ live lymphocytes are CD8+/HA Pentamer+ with a background stain of 0.03%.

The author commented that this study “confirms the novel ProImmune technology as a useful and superior tool for revealing potential immunogenic epitopes”, and also highlighted the importance of validating results in parallel using functional assays, such as ELISpot and Pro5® Pentamer staining.

重点出版物

See how Baxter (now Baxalta) to have used ProImmune REVEAL® to understand the immunogenicity of human Factor VIII treatment:

Steinitz, K. et al . “CD4+ T-cell epitopes associated with antibody responses after intravenously and subcutaneously applied human FVIII in humanized hemophilic E17 HLA-DRB1*15:01 mice.” Blood (2012) [PubMedID:22394599]