Peptide Pool Screening
Peptide Pool Screening - Related Videos
13:37
Split-and-pool Synthesis and Characterization of Peptide Tertiary Amide Library
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Cited by 5 •
2014
Peptide tertiary amides (PTAs) are a superfamily of peptidomimetics that include but are not limited to peptides, peptoids and N-methylated peptides. Here we describe a synthetic method which combines both split-and-pool and sub-monomer strategies to synthesize a one-bead one-compound library of PTAs.
09:12
MISSION LentiPlex Pooled shRNA Library Screening in Mammalian Cells
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Cited by 11 •
2011
Here we use a human LentiPlex pooled library and traditional sequencing methods to identify gene targets promoting cell survival. We demonstrate how to set up and deconvolute a LentiPlex screen and validate the results.
11:15
Pooled shRNA Screen for Reactivation of MeCP2 on the Inactive X Chromosome
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Cited by 2 •
2018
We report a small hairpin RNA (shRNA) and next generation sequencing-based protocol for identifying regulators of X-chromosome inactivation in a murine cell line with firefly luciferase and hygromycin resistance genes fused to the methyl CpG binding protein 2 (MeCP2) gene on the inactive X chromosome.
00:09
Pooled CRISPR-Based Genetic Screens in Mammalian Cells
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Cited by 14 •
2019
CRISPR-Cas9 technology provides an efficient method to precisely edit the mammalian genome in any cell type and represents a novel means to perform genome-wide genetic screens. A detailed protocol discussing the steps required for the successful performance of pooled genome-wide CRISPR-Cas9 screens is provided here.
02:42
A Fluorogenic Peptide Cleavage Assay to Screen the Proteolytic Activity of Proteases
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2025
This video demonstrates an assay to screen for the proteolytic activity of proteases using fluorogenic peptides. The protease recognizes its cleavage site on the peptide, cleaving it and separating the quencher from the fluorophore, enabling its fluorescence emission. The fluorescence signal is detected and analyzed to check for the cleavage efficiency of different peptide variants.
FAQs
Overlapping peptides help map which portions of a larger protein are immunologically recognized. Because neighboring peptides share sequence, a response to a pool can point investigators toward an antigen-derived region recognized by immune cells. This design supports comparisons of cellular recognition across defined pathogen antigens and helps identify regions relevant to infection research and vaccine development.
These readouts provide complementary ways to detect antigen-specific T-cell activation. Cytokine release measures secreted immune signals, intracellular staining detects cytokine production within responding cells, and ELISpot assays provide another format for measuring cellular responses. Using these options allows investigators to evaluate whether defined peptide antigens stimulate immune activity in responder-cell samples.
Because the assay uses synthetic peptides representing defined protein antigens, infection studies can examine cellular recognition without requiring the complete pathogen in the test system. The approach applies to viruses, bacteria, and other pathogens, allowing researchers to focus measurements on selected antigenic targets while characterizing pathogen-directed immune responses.
A typical workflow selects synthetic, overlapping peptides from a protein antigen and combines them into a test pool. The pool is incubated with peripheral blood mononuclear cells or another responder-cell population. Investigators then measure antigen-specific T-cell activation through cytokine release, intracellular staining, or an ELISpot assay.
Researchers use this approach to evaluate cellular responses during infectious disease studies, vaccine development, and immune monitoring. It can reveal whether immune cells respond to defined pathogen antigens and can help identify immunogenic regions for further investigation. The same strategy also supports comparisons of responses across samples or patient groups.
Testing responder cells against defined peptide pools can show whether a person’s immune cells recognize selected antigens, supporting assessment of patient-specific cellular responses. In infection research, the method also helps characterize immune memory by examining antigen-directed T-cell activity in available cell samples. These results can inform immune monitoring and studies of prior antigen exposure.