中国药物筛选平台(ScreeningPlatforms)

an HIV enzyme immunoassay
(EIA) is the first and most basic
test to determine if an individual
is positive for a selected pathogen,
such as HIV. The test is performed
Assay Reagents
Assay Development And Optimization
Screen Platforms
HTS, uHTS
Screen Development And Validation
Assessment
Assay Transfer
Screen Automation And Optimization
Absorbance assays Adsorption assays
SPA plate
Fluorescence assays Precipitation (沉淀) assays Bead based assays Radioimmunoassays
Cell-free Biochemical Assays
3、体外（in vitro）筛选 （cell free）
采用系统 简单或复杂（酶反应、蛋白－蛋白相互作用、 膜受体－配体结合、可溶性受体－配体结合检测 实验）
特点 a 、被筛化合物易于直接作用于靶标； b、目标化合物作用靶标明确； c、明确的作用机理； d、易于发展便宜易得的靶标模型； e、适应新技术的发展，易于自动化。
in a 8 cm x 12 cm plastic plate
whichBaidu Nhomakorabeacontains an 8 x 12 matrix
of 96 wells, each of which is
about 1 cm high and 0.7 cm in
An ELISA plate
diameter.
Positive ELISA Test
Negative ELISA Test
HIV antigens pre-coated onto an ELISA plate
Patient serum containing antibodies. If the patient is HIV+, then this serum will contain antibodies to HIV, and those antibodies will bind to the HIV antigens on the plate.
Screening Compound Deck
Lead Generation
Screen Paradigm
III、筛选平台
•Being familiar with various screen platforms will be in an advantageous position to evaluate the best assay for the target screening with the identification of a target. (靶标）
In vitro cell-free biochemical assays
Homogeneous
Heterogeneous
Radioactive Non-radioactive
assays
assays
Radioactive assays
Non-radioactive assays
ELISA assays SPA bead Chromogenic assays Filtration assays
（1）Nonradioactive Heterogeneous Assays
Enzyme immunoassays (widely used in vitro assays)
ELISA （Enzyme Linked Immunosorbent Assay）
酶联免疫吸附试验
An HIV ELISA, sometimes called
•Understanding various screen platforms will help in arriving at the best screens with the available equipment and reagents. （设备和试剂）
Targets
Reagents and plate readers availability
中国药物筛选平台 （Screening Platforms）
•药物筛选在新药发现中的地位 •药物筛选的常见重要方法 •重要疾病治疗药物筛选方法举例 •展望
虚拟筛选技术 （计算机科学） 生物物理化学技术、组合化学 分子细胞生物学 天然产物化学（中草药资源） （后基因组时代药物筛选新模式）
Biological Target Selection
Biacore 3000，96 生物分子相互作用分析系统
Biacore S51，384
96
2. Assay techniques
依据：激活或抑制、 激动剂(activator)、抑制剂 (inhibitor)
方法:
（1）体外筛选（in vitro)（酶活、受体结合）
（cell-free） （2）细胞水平（cell-based) （Heterogeneous & Homogeneous types)
A. Heterogeneous Assays
多步筛选方法 multiple additions/incubations/washings/ transfers/filtrations/readings of the signal
特点
Labor intensive, complicated step, hard to automate
Assay Design
1、 Assay formats
20世纪70年代：低通量和单一试管筛选方法；
现在:
(1)多孔板筛选 （96－、384－、1536－，3456－，9600 孔板筛选技术相继出现）
(2) 平板读数（plate readers) 技术：荧光、发光、闪烁等 检测技术的发展。
Stackers holding several plates (10-40 plates).