HLA antibody affinity determination. From HLA‐specific monoclonal antibodies to donor HLA specific antibodies (DSA) in patient serum
Loading...
Authors
Hug, Melanie N.
Gygax, Daniel
Handschin, Joëlle
Kleiser, Marc
Vazquez, Noemi
Linnik, Janina
Author (Corporation)
Publication date
16.05.2023
Typ of student thesis
Course of study
Collections
Type
01A - Journal article
Editors
Editor (Corporation)
Supervisor
Parent work
HLA
Special issue
DOI of the original publication
Link
Series
Series number
Volume
102
Issue / Number
3
Pages / Duration
Patent number
Publisher / Publishing institution
Wiley
Place of publication / Event location
Edition
Version
Programming language
Assignee
Practice partner / Client
Abstract
Organs transplanted across donor‐specific HLA antibodies (DSA) are associated with a variety of clinical outcomes, including a high risk of acute kidney graft rejection. Unfortunately, the currently available assays to determine DSA characteristics are insufficient to clearly discriminate between potentially harmless and harmful DSA. To further explore the hazard potential of DSA, their concentration and binding strength to their natural target, using soluble HLA, may be informative. There are currently a number of biophysical technologies available that allow the assessment of antibody binding strength. However, these methods require prior knowledge of antibody concentrations. Our objective within this study was to develop a novel approach that combines the determination of DSA‐affinity as well as DSA‐concentration for patient sample evaluation within one assay. We initially tested the reproducibility of previously reported affinities of human HLA‐specific monoclonal antibodies and assessed the technology‐specific precision of the obtained results on multiple platforms, including surface plasmon resonance (SPR), bio‐layer interferometry (BLI), Luminex (single antigen beads; SAB), and flow‐induced dispersion analysis (FIDA). While the first three (solid‐phase) technologies revealed comparable high binding‐strengths, suggesting measurement of avidity, the latter (in‐solution) approach revealed slightly lower binding‐strengths, presumably indicating measurement of affinity. We believe that our newly developed in‐solution FIDA‐assay is particularly suitable to provide useful clinical information by not just measuring DSA‐affinities in patient serum samples but simultaneously delivering a particular DSA‐concentration. Here, we investigated DSA from 20 pre‐transplant patients, all of whom showed negative CDC‐crossmatch results with donor cells and SAB signals ranging between 571 and 14899 mean fluorescence intensity (MFI). DSA‐concentrations were found in the range between 11.2 and 1223 nM (median 81.1 nM), and their measured affinities fall between 0.055 and 24.7 nM (median 5.34 nM; 449‐fold difference). In 13 of 20 sera (65%), DSA accounted for more than 0.1% of total serum antibodies, and 4/20 sera (20%) revealed a proportion of DSA even higher than 1%. To conclude, this study strengthens the presumption that pre‐transplant patient DSA consists of various concentrations and different net affinities. Validation of these results in a larger patient cohort with clinical outcomes will be essential in a further step to assess the clinical relevance of DSA‐concentration and DSA‐affinity.
Keywords
Subject (DDC)
500 - Naturwissenschaften und Mathematik
Event
Exhibition start date
Exhibition end date
Conference start date
Conference end date
Date of the last check
ISBN
ISSN
2059-2302
2059-2310
2059-2310
Language
English
Created during FHNW affiliation
Yes
Strategic action fields FHNW
Publication status
Published
Review
Peer review of the complete publication
Open access category
Hybrid
License
Citation
HUG, Melanie N., Sabrina KELLER, Talea MARTY, Daniel GYGAX, Dominik MEINEL, Peter SPIES, Joëlle HANDSCHIN, Marc KLEISER, Noemi VAZQUEZ, Janina LINNIK, Rico BUCHLI, Frans CLAAS, Sebastiaan HEIDT, Cynthia S. M. KRAMER, Suzanne BEZSTAROSTI, Jar‐How LEE, Stefan SCHAUB und Gideon HÖNGER, 2023. HLA antibody affinity determination. From HLA‐specific monoclonal antibodies to donor HLA specific antibodies (DSA) in patient serum. HLA. 16 Mai 2023. Bd. 102, Nr. 3. DOI 10.1111/tan.15047. Verfügbar unter: https://doi.org/10.26041/fhnw-5377