CORE
🇺🇦
make metadata, not war
Services
Services overview
Explore all CORE services
Access to raw data
API
Dataset
FastSync
Content discovery
Recommender
Discovery
OAI identifiers
OAI Resolver
Managing content
Dashboard
Bespoke contracts
Consultancy services
Support us
Support us
Membership
Sponsorship
Community governance
Advisory Board
Board of supporters
Research network
About
About us
Our mission
Team
Blog
FAQs
Contact us
PeptiCHIP : A Microfluidic Platform for Tumor Antigen Landscape Identification
Authors
Gabriella Antignani
Rui M. M. Branca
+26 more
Cristian Capasso
Vincenzo Cerullo
Jacopo Chiaro
Masoumeh Eshaghi
Antonio Federico
Michaela Feodoroff
Sara Feola
Manlio Fusciello
Dario Greco
Mikaela Grönholm
Olga L. Gurvich
Markus Haapala
Firas Hamdan
Tuija Kekarainen
Satu Koskela
Janne Lehtiö
Beatriz Martins
Jukka Partanen
Karita Peltonen
Vilja Pietiäinen
Antti Rannikko
Salvatore Russo
Tiina M. Sikanen
Sari M. Tähkä
Seppo Ylä-Herttuala
Erkko Ylösmäki
Publication date
1 January 2021
Publisher
Doi
Cite
Abstract
Publisher Copyright: © 2021 The Authors. Published by American Chemical Society.Identification of HLA class I ligands from the tumor surface (ligandome or immunopeptidome) is essential for designing T-cell mediated cancer therapeutic approaches. However, the sensitivity of the process for isolating MHC-I restricted tumor-specific peptides has been the major limiting factor for reliable tumor antigen characterization, making clear the need for technical improvement. Here, we describe our work from the fabrication and development of a microfluidic-based chip (PeptiCHIP) and its use to identify and characterize tumor-specific ligands on clinically relevant human samples. Specifically, we assessed the potential of immobilizing a pan-HLA antibody on solid surfaces via well-characterized streptavidin-biotin chemistry, overcoming the limitations of the cross-linking chemistry used to prepare the affinity matrix with the desired antibodies in the immunopeptidomics workflow. Furthermore, to address the restrictions related to the handling and the limited availability of tumor samples, we further developed the concept toward the implementation of a microfluidic through-flow system. Thus, the biotinylated pan-HLA antibody was immobilized on streptavidin-functionalized surfaces, and immune-affinity purification (IP) was carried out on customized microfluidic pillar arrays made of thiol-ene polymer. Compared to the standard methods reported in the field, our methodology reduces the amount of antibody and the time required for peptide isolation. In this work, we carefully examined the specificity and robustness of our customized technology for immunopeptidomics workflows. We tested this platform by immunopurifying HLA-I complexes from 1 × 106 cells both in a widely studied B-cell line and in patients-derived ex vivo cell cultures, instead of 5 × 108 cells as required in the current technology. After the final elution in mild acid, HLA-I-presented peptides were identified by tandem mass spectrometry and further investigated by in vitro methods. These results highlight the potential to exploit microfluidics-based strategies in immunopeptidomics platforms and in personalized immunopeptidome analysis from cells isolated from individual tumor biopsies to design tailored cancer therapeutic vaccines. Moreover, the possibility to integrate multiple identical units on a single chip further improves the throughput and multiplexing of these assays with a view to clinical needs.Peer reviewe
Similar works
Full text
Open in the Core reader
Download PDF
Available Versions
Trepo - Institutional Repository of Tampere University
See this paper in CORE
Go to the repository landing page
Download from data provider
oai:trepo.tuni.fi:10024/135167
Last time updated on 24/12/2021
Helsingin yliopiston digitaalinen arkisto
See this paper in CORE
Go to the repository landing page
Download from data provider
oai:helda.helsinki.fi:10138/33...
Last time updated on 21/12/2021