Publication: Comparison of the established
standard complement-dependent cytotoxicity
and flow cytometric crossmatch assays with a
novel ELISA-based HLA crossmatch procedure
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Date
2006
Authors
Altermann, W.W. ; Seliger, B. ; Sel, S. ; Wendt, D.
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Publisher
Murcia : F. Hernández
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DOI
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info:eu-repo/semantics/article
Description
Abstract
The detection of donor-specific anti-HLA
antibodies by standard procedures such as complementdependent
cytotoxicity assay (CDC) or flow cytometric
(FACS) analysis is limited by its low sensitivity and the
quality of the donor cells. Therefore, an ELISA-based
technique was employed using solid phase-immobilized
monoclonal antibodies to capture HLA class I or class II
molecules of the donor, respectively. In this HLA class I
and class II antibody monitoring system (AMS) the
donor-specific anti-HLA antibodies from the sera of
recipients bind to the HLA molecules of the donor which
have been immobilized by monoclonal antibodies (mAb)
recognizing non-polymorphic epitopes. Upon binding of
donor-specific anti-HLA antibodies they are recognized
by secondary enzyme-conjugated anti-human
immunoglobulin (Ig) antibodies. A newly established
modification of the standard protocol allows the
differentiation between bound antibodies of the IgG and
IgM isotype. Furthermore, this assay was adapted for
investigating small amounts of solid tissue of donors
from whom no other cells (e.g. from blood) were
available. We here provide an overview of the classical
crossmatch methods with their advantages and limits. In
addition, the design of the novel AMS-ELISA is
described in terms of quality and sensitivity of the
approach using exemplary cases of different application.
The selected cases show that the AMS-ELISA represents
a valuable tool for the post-transplantation monitoring of
donor-specific anti-HLA antibodies during reaction
crisis, after transfusion reactions and in particular cases
of tissue transplantations lacking single cells.
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