What Is High Throughput Screening?

High-throughput screening (HTS) technology refers to a method based on molecular and cell-level experiments, a microplate as a carrier of experimental tools, an automated operating system to perform the test process, and a sensitive and rapid detection instrument to collect Experimental result data, computer analysis and processing of experimental data, detection of tens of millions of samples at the same time, and the corresponding database to support the operation of the technical system, it has the characteristics of trace, fast, sensitive and accurate. In short, you can get a lot of information through one experiment and find valuable information from it.

High-throughput screening

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High throughput screening (HTS) technology refers to

In high-throughput screening, tens of millions of

Full use of medicinal resources due to high-throughput screening

experimental method

High-throughput screening experimental methods Molecular-level and cell-level experimental methods (or screening models) are the technical basis for achieving high-throughput screening of drugs. Because high-throughput screening of drugs requires processing a large number of samples simultaneously, the experimental system must be miniaturized, and these miniaturized experimental methods should be established based on new scientific research results. The Fourth Military Medical University on Thursday said that the experimental methods of high-throughput screening models of drugs can be divided into the following categories based on their biological characteristics: receptor binding assays; enzyme activity assays; cell molecular assays; Method; metabolite assay; gene product assay. These experimental methods have been widely used in high-throughput screening of drugs.

Featured utility

The characteristics of high-throughput screening High-throughput screening technology is a new technology system formed by combining a variety of technical methods. It uses a microplate as an experimental tool carrier, and uses an automated operating system to perform the experimental process. It is sensitive and fast. The testing instrument collects experimental data, and analyzes and processes thousands of sample data with a computer, so as to obtain scientific and accurate experimental results and characteristic utility. British scholar AlanD research suggests that a laboratory adopts traditional methods and uses more than 20 drug targets to screen only 75,000 samples in one year. In the early stage of the development of high-throughput screening technology in 1997, more than 100 targets were used. One million samples can be screened; after the high-throughput screening technology was further improved in 1999, the daily screening volume was up to 100,000 compounds.

FLIPR fluorescence detection

In recent years, researchers in the United States and the United Kingdom have been working on optical measurement techniques in high-throughput screening and testing. They have worked hard on optical measurement methods and established a large number of non-isotopic labeling assays, such as screening protein tyrosine by spectrophotometry Acid kinase inhibitors and tissue plasminogen activators have all been successful.

Radioactive detection technology American scholar GanieSM applied radiometric methods, especially affinity scintillation (SPA) detection methods in the research of high-throughput drug screening, to develop the sample size experiments performed on 96-well plates. This method has high sensitivity and specificity, which promotes the realization of high-throughput drug screening, but it has environmental pollution problems.

Fluorescence detection technology American scholar GiulianokA research believes that the FLIPR (fluorometricimagingreadet) fluorescence detection method can be used to simultaneously measure the intensity and change of fluorescence in a short period of time. , Is an ideal and efficient detection method. At the same time, FDSS (Functional Drug Screening System) is used for real-time multi-channel fluorescence detection, and 96-well plates, 384-well plates, and 1536-well plates are loaded at one time to achieve real-time fluorescence intensity signal detection. The following applications can be used:

GPCR calcium influx assays

--All calcium wash or non-washing kits (Fluo-4 or Fura-2 based), Premo Cameleon, Aequorin

· Ion Channel assays

--FMP, VSP, potassium channel (FluxOR ), sodium channel (SBFI and ANG-2), chloride channel (YFP)

· Enzymatic assays

--Prolyl isomerase, GTPase, Kvb

Transporter assays

· CytoStar-T

· Circadian clock

Mosquito Odorant Receptors

Light activated receptor or channel assays

Multifunctional microplate detection system is a 1536-well plate high-throughput multifunctional microplate detection system developed by the School of Pharmacy, Xi'an Jiaotong University. It is currently an internationally advanced high-throughput detection system, which can further increase the screening capacity. The hospital was put into use.

AlphaScreen detection method

AlphaScreen assay is widely used and has many advantages to optimize the effectiveness of drug screening tests. Due to the limitations of its operating hardware, although this experimental method has been developed for several years, most researchers have not known or widely used it.

The research focus of the Chemical Biology Research Project is to screen high-throughput detection of protein-protein interactions performed on our platform. For this purpose, we use AlphaScreen assay (Amplified Luminescence Proximity Homogeneous Assay). As its name implies, ALPHA screens are based on luminous proximity detection.

The main advantage of AlphaScreen technology lies in the wide range of substances to be measured, from small molecules to large complexes; homogeneous system, fast, stable, and higher sensitivity; AlphaScreen detection also does not require the introduction of fluorescent labels, which avoids steric hindrance affecting biological molecules. Combined with each other; can be used to detect biological crude extracts such as cell lysates, serum, plasma, body fluids, etc. without affecting the reading effect.

The main limitation of AlphaScreen technology is that the reaction system is sensitive to strong light or long-term indoor light. Second, the capture of monomeric oxygen molecules by some compounds will reduce the light signal. The pearl bleaching effect of the donor makes the signal detection better in a single shot. . Similar to ECL and FMAT technologies, AlphaScreen also requires high-energy lasers; compared to other technologies, AlphaScreen has requirements for detection instrument platforms.

Technological progress

The advantages of high-throughput screening of drugs in China are firstly that the compounds are from a wide range of sources and are mostly natural; secondly, the purpose of screening compounds for biological activity is clear, and fewer compounds are synthesized without purpose; third, traditional Chinese drugs provide screening studies A huge resource library can be extracted and screened for new compounds from traditional Chinese medicine. These advantages provide a solid foundation for high-throughput screening of drugs.

The initial scale of high-throughput screening of drugs in China: High-throughput screening of drugs started late in China and is not standardized. In recent years, China has carried out the integrated and large-scale infrastructure construction of outbound and inward linking, and has achieved initial results. In 1996, the Chinese Academy of Medical Sciences introduced the first Biomek2000 experimental automated workstation in China; in 1998, it introduced the first Topcount micro-scintillation counter in the country to miniaturize and automate technologies such as radioligand experiments and radioimmunity experiments. The Shanghai Institute of Materia Medica and the Beijing Academy of Military Medical Sciences have set up specialized drug screening agencies and started to engage in large-scale screening. The Cell Membrane Chromatography (CMC) pioneered by Professor He Langchong, School of Pharmacy, Xi'an Jiaotong University, provides a highly selective, highly specific and efficient screening method for high-throughput screening of compounds in vitro. CMC has been successfully used in the study of calcium ion antagonist receptor ligand binding reactions. Currently, high-throughput screening of cardiovascular chemical synthetic drugs and the search for effective parts and components of traditional Chinese medicines are underway. The molecular biological method will be used to establish an automated CMC screening system to promote the comprehensive development of China's high-throughput screening technology for drugs.

High-throughput screening technology is an important research topic in the field of drug screening. In recent years, although its research and application have achieved great development, there are still many problems, such as the relationship between the screening results of in vitro models and the overall pharmacological effect. ; Evaluation criteria for high-throughput screening models, and research and discovery of new drug targets. With the advancement of medicine, high-throughput screening technology will definitely open up a wider space in the development of innovative drugs.