High-content screening platform for high-throughput, no-clean detection of antibody binding

Screening is the beginning of the antibody drug development process. Screening for specific antibodies from supernatants secreted by thousands of monoclonal cell lines has been a core area of ​​initial drug development. For the antibody drug development market that is now fully accelerated, it is important to complete the screening work quickly and efficiently.

ELISA (for protein antigens) and flow cytometry (for cell membrane surface antigens) are still widely used screening methods, but they all have their own limitations. The ELISA screening method is aimed at expressing the purified protein antigen, and the conformation is different from that in the cell, resulting in the screening of high affinity candidate antibodies may be false positive, and the ELISA method requires cumbersome washing and incubation, greatly The workload is increased; the flow cytometry screening method is directed to the cell membrane surface antigen, but it takes a lot of cells, and the process of cell digestion by trypsin also has an effect on the surface antigen.

Our platform effectively solves these problems based on the high-throughput antibody screening platform established by PerkinElmer's latest Operetta CLS high-content analyzer. Equipped with a confocal module, the instrument can directly perform fluorescence imaging on 96, 384 or 1536 well-labeled adherent cells containing high concentrations of fluorescent secondary antibodies without high background fluorescence, thereby enabling cell surface antibody-binding fluorescence. High-speed automated no-clean detection of signals (this method has been repeatedly verified by our rigorous internal experiments). Compared to ELISA or flow cytometry, high-content analysis techniques can significantly reduce reagent consumables, labor, and time costs in antibody screening.

The operation procedure of this screening platform is simple and no washing step is required in the whole process: 1. Select the cell plate to be detected; 2. Add the antibody to be detected for incubation; 3. Add the secondary antibody to incubate; 4. Analyze the result by OperettaCLS.

Figure 1. Schematic diagram of the operational flow of the Operetta CLS high-content screening platform.

For the Immuno-Oncology related targets, we have constructed their over-expressing stable cell lines, and the target well plates that are now available for service are shown in Figure 2.

Figure 2. Construction of overexpressing stable cell lines for Immuno-Oncology related targets. A, completed and under construction targets; B, CHO-K1 (PD-L1) monoclonal flow detection results; C, monoclonal tracking verification of overexpressing cell lines by Operetta CLS high content analyzer .

After selecting the target of your favorite, just give us the supernatant to be tested, and you can get the experimental results with the truth as shown in Figure 3. The detection limit can reach 5 ng/ml.

Figure 3. Operetta high-content screening platform detects Anti-PD-L1 antibody binding ability.

At present, the service has been fully opened to the public, and teachers who are interested inside and outside are welcome to consult.

Contact:

Yan Kefeng ( / 18651111318)

Zheng Kexiao ( / 18015595729)

For more information on high-intensity instruments, please click on the instrument page to find out:
Http://?equipid=4269109&division=1213

In early April, PerkinElmer and the Nano-Biochemical Platform of the Suzhou Institute of Nanoscience of the Chinese Academy of Sciences jointly established a cooperative laboratory. Related ppt courseware can be downloaded at the following address: