During the cell line development process hundreds to thousands of clones have to be screened to find the most promising, i.e. well growing clones that are also good producers for further development. The process of single cell cloning is usually performed in 96-well formats and increasingly also in 384-well plates. Thus, there is limited sample volume available for analysis and it is desirable to get the results as quickly as possible.
PAIA BIOTECH has taken on this challenge and their scientific team has developed a new ultra-fast-low-volume assay for IgG-quantification directly from the supernatant after single cell cloning!
This new technology gives SYNENTEC the opportunity, not only to determine the clonality and to support the process optimization with our high throughput imaging systems, but also to find in combination with the PAIAplate the high producer which also still grows fast and as quickly as possible. One full 384-well PAIAplate is processed in less than one hour and allows reliable identification of high producers in an automation-friendly workflow.
Protein A coated capture beads, analyte and fluorescence markers are incubated in the wells of a 384-well PAIAplate. After sedimentation of the beads with the bound analyte-marker-complexes, the remaining unbound fluorescence marker is measured with the NYONE® or CELLAVISTA®.
- High throughput in 384 well plates
- processing time for full plate < 1 hour
- Low sample consumption
- Simple protocols in single-use plates
- no washing , no blocking, no regeneration
- Cost-efficient (No capital expenditures)
- runs on fluorescence microscopes
Running PAIA IgG quantification assays on the CELLAVISTA® and NYONE® imagers allow the reliable identification of high producers with a very limited amount of samples from cell culture supernatants and high throughput. The combination of PAIA assays and SYNENTEC's imaging systems enables to monitor monoclonality and cell growth plus the workflow is amenable to automation and offers a high throughput which can be increased to up to four 384-well plates per hour if several shakers are used in parallel.
PAIA Biotech has developed a software tool for the rapid generation of calibration curves, calculation of sample concentrations and result exports. The data generated with our YT-analysis software® can be exported as csv-files and can be imported into PAIAs software tool. In this software tool method templates facilitate the processing of recurrent plate layouts. They contain a predefined layout with the positions of standards, controls and samples as well as their concentration and dilution factors.
The measured fluorescence intensities are copied from the instrument and pasted into the tool, calibration curves are generated, and heat maps provide a quick result overview. Different report and export functions for QC data, results and graphs are implemented.