WesJess Simple 全自动定量western blot 病原微生物感染研究案例

一、检测病原微生物感染蛋白

TM6SF2 Promotes Lipidation and Secretion of Hepatitis C Virus in Infected Hepatocytes, Gastroenterology Volume 155, Issue 6, December 2018, Pages 1923-1935.e8.

二、病原微生物高免疫原性蛋白筛选

       (诊断和疫苗开发靶点)

       案例1. 结核杆菌高免疫原性蛋白筛选

Analysis of the Antigenic Properties of Membrane Proteins of Mycobacterium tuberculosis, Scientific Reports | (2019) 9:3042.

案例2. 埃博拉病毒病毒特异性IgG和IgM检测

Qualitative Profiling of the Humoral Immune Response Elicited by rVSV-DG-EBOV-GP Using a Systems Serology Assay, Domain Programmable Arrays,Cell Reports 24, 1050–1059, July 24, 2018.

三、病原微生物抗体验证实验

An Automated Immunoblot Method for Detection of IgG Antibodies to Hepatitis C Virus: a Potential Supplemental Antibody Confirmatory Assay, Journal of Clinical Microbiology, March 2019 Volume 57 Issue 3 e01567-18.

四、 病人样本蛋白定量检测

1. 支气管-肺灌洗液（甲流）

Quantitative protein analysis

The Wes system was used to identify and quantify BPIFA1 protein in broncho-alveolar lavage and lung samples (ProteinSimple) and rabbit anti-mBPIFA1 as a primary antibody. The area of BPIFA1 peak, in relation to the standard curve, was determined and virtual blot-like images were generated using Compass software (ProteinSimple).

An innate defense peptide BPIFA1/SPLUNC1 restricts influenza A virus infection, Mucosal Immunology volume 11, pages 71–81(2018).

2. 脑脊液

（肺炎链球菌脑炎病人）

Quantitative western blotting

Automatic western blots were performed using a Wes automated system (ProteinSimple, California, USA). Purified recombinant proteins were used as calibration standards. Serial dilutions of both sample and standard were used to determine the linear dynamic range of the assay. Additionally, the optimal concentration of each antibody for use in the Wes system was determined, as this can differ from that used in traditional western blot. Samples (20 SPP, 15 hospital controls and 5 healthy controls) were mixed with a 5x sample buffer containing SDS, DTT and fluorescent molecular weight standards and heated at 95 °C for 5 min and then, loaded onto a plate prefilled with stacking and separation matrices, along with blocking and wash buffers, antibody solutions and detection reagents. Default settings were used for the analysis.

Quantitative Proteomics of Cerebrospinal Fluid in Paediatric Pneumococcal Meningitis, Scientific Reports volume 7, (2017).

Wes/Jess 全自动定量Western Blot特点

         n  全程自动化，无需检测人员值守

         n  从样本到结果，全程3小时

         n  标准曲线定量检测病人样本靶蛋白浓度

         n  微量样本

         n  数据可追溯

         n  自动数据计算存储