PMID- 36179215
OWN - NLM
STAT- MEDLINE
DCOM- 20221026
LR  - 20231130
IS  - 1520-6882 (Electronic)
IS  - 0003-2700 (Print)
IS  - 0003-2700 (Linking)
VI  - 94
IP  - 42
DP  - 2022 Oct 25
TI  - High-Throughput, Quantitative Analysis of Peptide-Exchanged MHCI Complexes by 
      Native Mass Spectrometry.
PG  - 14593-14602
LID - 10.1021/acs.analchem.2c02423 [doi]
AB  - Immune monitoring in cancer immunotherapy involves screening CD8+ T-cell 
      responses against neoantigens, the tumor-specific peptides presented by Major 
      histocompatibility complex Class I (MHCI) on the cell surface. High-throughput 
      immune monitoring requires methods to produce and characterize small quantities 
      of thousands of MHCI-peptide complexes that may be tested for a patient's T-cell 
      response. MHCI synthesis has been achieved using a photocleavable peptide that is 
      exchanged by the neoantigen; however, assays that measure peptide exchange 
      currently disassemble the complex prior to analysis horizontal line precluding direct molecular 
      characterization. Here, we use native mass spectrometry (MS) to profile intact 
      recombinant MHCI complexes and directly measure peptide exchange. Coupled with 
      size-exclusion chromatography or capillary-zone electrophoresis, the assay 
      identified all tested human leukocyte antigen (HLA)/peptide combinations in the 
      nanomole to picomole range with minimal run time, reconciling the synthetic and 
      analytical requirements of MHCI-peptide screening with the downstream T-cell 
      assays. We further show that the assay can be "multiplexed" by measuring exchange 
      of multiple peptides simultaneously and also enables calculation of Vc(50), a 
      measure of gas-phase stability. Additionally, MHCI complexes were fragmented by 
      top-down sequencing, demonstrating that the intact complex, peptide sequence, and 
      their binding affinity can be determined in a single analysis. This screening 
      tool for MHCI-neoantigen complexes represents a step toward the application of 
      state-of-the-art MS technology in translational settings. Not only is this assay 
      already informing on the viability of immunotherapy in practice, the platform 
      also holds promise to inspire novel MS readouts for increasingly complex 
      biomolecules used in the diagnosis and treatment of disease.
FAU - Schachner, Luis F
AU  - Schachner LF
AUID- ORCID: 0000-0001-6157-0937
AD  - Department of Microchemistry, Proteomics and Lipidomics, Genentech Inc., South 
      San Francisco, California 94080, United States.
FAU - Phung, Wilson
AU  - Phung W
AD  - Department of Microchemistry, Proteomics and Lipidomics, Genentech Inc., South 
      San Francisco, California 94080, United States.
FAU - Han, Guanghui
AU  - Han G
AD  - BGI Americas, San Jose, California 95134, United States.
FAU - Darwish, Martine
AU  - Darwish M
AD  - Department of Protein Chemistry, Genentech Inc., South San Francisco, California 
      94080, United States.
FAU - Bell, Ashley
AU  - Bell A
AD  - 908 Devices, Carrboro, North Carolina 27510, United States.
FAU - Mellors, J Scott
AU  - Mellors JS
AUID- ORCID: 0000-0002-6658-3961
AD  - 908 Devices, Carrboro, North Carolina 27510, United States.
FAU - Srzentic, Kristina
AU  - Srzentic K
AD  - Thermo Fisher Scientific, San Jose, California 95134, United States.
FAU - Huguet, Romain
AU  - Huguet R
AD  - Thermo Fisher Scientific, San Jose, California 95134, United States.
FAU - Blanchette, Craig
AU  - Blanchette C
AD  - Department of Protein Chemistry, Genentech Inc., South San Francisco, California 
      94080, United States.
FAU - Sandoval, Wendy
AU  - Sandoval W
AUID- ORCID: 0000-0002-4672-0762
AD  - Department of Microchemistry, Proteomics and Lipidomics, Genentech Inc., South 
      San Francisco, California 94080, United States.
LA  - eng
GR  - T32 GM105538/GM/NIGMS NIH HHS/United States
PT  - Journal Article
DEP - 20220930
PL  - United States
TA  - Anal Chem
JT  - Analytical chemistry
JID - 0370536
RN  - 0 (Histocompatibility Antigens Class I)
RN  - 0 (Peptides)
RN  - 0 (HLA Antigens)
RN  - 0 (Antigens, Neoplasm)
SB  - IM
MH  - Humans
MH  - *Histocompatibility Antigens Class I/metabolism
MH  - *Peptides/chemistry
MH  - Mass Spectrometry
MH  - HLA Antigens
MH  - Antigens, Neoplasm
PMC - PMC9607865
COIS- The authors declare the following competing financial interest(s): LS, WP, MD, CB 
      and WS are employed by Genentech, Inc., a for-profit company that produces and 
      markets therapeutics.AB and SM are employed by 908Devices, GH by BGI, and KS and 
      RH by Thermo Fisher Scientific, for-profit companies that manufacture and sell 
      mass spectrometry equipment.
EDAT- 2022/10/01 06:00
MHDA- 2022/10/27 06:00
PMCR- 2022/10/27
CRDT- 2022/09/30 15:52
PHST- 2022/10/01 06:00 [pubmed]
PHST- 2022/10/27 06:00 [medline]
PHST- 2022/09/30 15:52 [entrez]
PHST- 2022/10/27 00:00 [pmc-release]
AID - 10.1021/acs.analchem.2c02423 [doi]
PST - ppublish
SO  - Anal Chem. 2022 Oct 25;94(42):14593-14602. doi: 10.1021/acs.analchem.2c02423. 
      Epub 2022 Sep 30.