Protein capture reagents program

Protein Capture Reagents Program grantee Dr. Common Fund programs are strategic investments that achieve a set of high-impact goals within a year timeframe. At the conclusion of each program, deliverables will transition to other sources of support or use within the scientific community.

Please note that since the Protein Capture Reagents program is no longer supported by the Common Fund, the program website is being maintained as an archive and will not be updated on a regular basis.

The program has developed new resources and tools to understand the critical role the multitude of cellular proteins play in development, health, and disease. These resources will support a wide-range of research and clinical applications by enabling the isolation and tracking of proteins of interest. The program generated an array of monoclonal antibodies to transcription factor targets in addition to a host of recombinant antibodies.

Users may need to further optimize protocols for reagents of interest according to their specific assay needs. CDI production center exploits a unique human proteome technology, the HuProt array with almost 20K spotted protein isoforms corresponding to The Rutgers antigen production group is responsible for designing, expressing, purifying, and shipping protein domain antigens to the Protein Capture Reagents Program antibody production John Chaput and Dr.

NuPromers are produced via a novel technology that combines a rigid DNA scaffold with two peptides that have affinity against a specific protein target. They applied this technology toward creating a relatively inexpensive and high throughput pipeline.

Andreas Pluckthun at the University of Zurich, and Dr. Michael Weiner at Illumina formed a research collaboration to improve screening and evaluation technologies currently used to generate affinity reagents.

This group compared three types of recombinant protein scaffolds as affinity reagents single-chain fragments of variable regions scFv , ankyrin repeat proteins, and fibronectin type III monobodies and two types of display technologies phage and ribosomal against the same set of human proteins and worked to optimize a pipeline for producing recombinant monoclonal antibodies.

This is a novel and ground-breaking approach for selection in phage display, where the generation of an emulsion phase allows the compartmentalization of each phage with the antigen linked to beads. Lloyd Smith, Dr. With validated reagents, researchers would have increased confidence when picking a reagent to use in their own studies and in reproducing the results of others. PCRP began by piloting a pipeline to generate reagents targeting a subset of proteins, transcription factors, before scaling to the entire human proteome.

The program produced and validated a collection of approximately antibodies capable of targeting around human transcription factors. However, the biomedical community expressed a need for additional validation of these antibodies under specific experimental conditions to encourage broader use of the reagents.

They also validated a subset of the antibodies in several other popular experimental approaches for detecting interactions between transcription factors and DNA, in both cell-free and cell-based assays.

Large-scale validation of antibodies as performed in this study is rare and therefore had not been performed in ChIP before. This work demonstrated the importance of experimental approach-specific validation to aid researchers in their antibody selection.

It also showed how validation criteria for reagents like antibodies that recognize other molecular structures can be applied to a particular experimental technique, including consideration of how experimental variations may affect outcomes. The researchers have made their data publicly available to inspire broader use of the PCRP antibodies and encourage further research on antibody validation.

While PCRP ultimately did not produce reagents to target the entire human proteome, the program did identify important lessons learned while providing validated protein-targeting reagents in an area that previously lacked a substantiative number of them. An important finding is that large-scale generation of reagents to target proteins is possible, but high-throughput and systematic validation of these reagents may not be feasible nor necessary.

Validation of reagents performed by individual labs for their own specific uses may be a better way to enhance rigor and reproducibility in biomedical research. Genome Research. Epub Aug Science Advances.

Epub Nov