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An ultra-high-throughput screen for the evaluation of peptide HLA-Binder interactions.


ABSTRACT: Peptide human leukocyte antigen (pHLA) targeting therapeutics like T-cell receptor based adoptive cell therapy or bispecific T cell engaging receptor molecules hold great promise for the treatment of cancer. Comprehensive pre-clinical screening of therapeutic candidates is important to ensure patient safety but is challenging because of the size of the potential off-target space. By combining stabilized peptide-receptive HLA molecules with microarray printing and screening, we have developed an ultra-high-throughput screening platform named ValidaTe that enables large scale evaluation of pHLA-binder interactions. We demonstrate its potential by measuring and analyzing over 30.000 binding curves for a high-affinity T cell Engaging Receptor towards a large pHLA library. Compared to a dataset obtained by conventional bio-layer interferometry measurements, we illustrate that a massively increased throughput (over 650 fold) is obtained by our microarray screening, paving the way for use in pre-clinical safety screening of pHLA-targeting drugs.

SUBMITTER: Kramer S 

PROVIDER: S-EPMC10066226 | biostudies-literature | 2023 Mar

REPOSITORIES: biostudies-literature

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An ultra-high-throughput screen for the evaluation of peptide HLA-Binder interactions.

Krämer Stefan S   Moritz Andreas A   Stehl Luca L   Hutt Meike M   Hofmann Martin M   Wagner Claudia C   Bunk Sebastian S   Maurer Dominik D   Roth Günter G   Wöhrle Johannes J  

Scientific reports 20230331 1


Peptide human leukocyte antigen (pHLA) targeting therapeutics like T-cell receptor based adoptive cell therapy or bispecific T cell engaging receptor molecules hold great promise for the treatment of cancer. Comprehensive pre-clinical screening of therapeutic candidates is important to ensure patient safety but is challenging because of the size of the potential off-target space. By combining stabilized peptide-receptive HLA molecules with microarray printing and screening, we have developed an  ...[more]

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