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Use of a recombinant deacetylase to convert A1 red blood cells to the acquired B phenotype for quality control purposes

Hagman, Jennifer Ricci LU ; Rahfeld, Peter ; Withers, Stephen G ; Kizhakkedathu, Jayachandran N ; Olsson, Martin L LU orcid and Storry, Jill R LU (2023) In Blood Transfusion
Abstract
Background - Correct blood group typing is the cornerstone of transfusion medicine. In patients whose gastrointestinal wall is compromised, bacterial enzymes can cause deacetylation of blood group A, converting N-acetylgalactosamine to galactosamine, which resembles the B-defining galactose. This acquired B (acqB) phenomenon, first described 1959, can lead to blood grouping errors. Herein, we explore the possibility of producing acqB red blood cells (RBCs) by enzymatic conversion, for quality control purposes.
Materials and methods - A1 RBCs along with group B and O control RBCs were subjected to enzymatic digestion by bacterially derived deacetylase, FpGalNAcDeAc. RBCs were tested with monoclonal anti-A, anti-B, acquired-B-reactive... (More)
Background - Correct blood group typing is the cornerstone of transfusion medicine. In patients whose gastrointestinal wall is compromised, bacterial enzymes can cause deacetylation of blood group A, converting N-acetylgalactosamine to galactosamine, which resembles the B-defining galactose. This acquired B (acqB) phenomenon, first described 1959, can lead to blood grouping errors. Herein, we explore the possibility of producing acqB red blood cells (RBCs) by enzymatic conversion, for quality control purposes.
Materials and methods - A1 RBCs along with group B and O control RBCs were subjected to enzymatic digestion by bacterially derived deacetylase, FpGalNAcDeAc. RBCs were tested with monoclonal anti-A, anti-B, acquired-B-reactive anti-B (ES4 clone), and a panel of donor plasmas. Standard serological techniques were used. RBCs were subsequently frozen by a glycerolisation method, stored at −80 oC, and thawed for testing to evaluate stability.
Results - Specific deacetylation of A antigen was achieved at both enzyme concentrations. Enzymatically modified A1 RBCs reacted 1-3+ with clone ES4 but not with other anti-B reagents. Group B and O RBC controls reacted as expected. Crossmatch testing revealed 1-3+ reactions in 17/26 group A plasma with modified RBCs but not with untreated controls. Furthermore, 2/3 group AB plasmas reacted strongly with the modified RBCs. The acqB phenotype was maintained upon freezing/thawing.
Discussion - Using a recombinant deacetylase, we demonstrated the feasibility of creating acqB RBCs from donor A1 RBCs, thus providing a reagent for quality assurance of monoclonal anti-B. These RBCs can be frozen once treated, providing a reliable source of this unusual but clinically important phenotype. (Less)
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author
; ; ; ; and
organization
publishing date
type
Contribution to journal
publication status
epub
subject
in
Blood Transfusion
publisher
Simiti Servizi SRL
external identifiers
  • scopus:85186962068
  • pmid:37847210
ISSN
1723-2007
DOI
10.2450/BloodTransfus.584
language
English
LU publication?
yes
id
5a30b44a-386b-4580-985e-116870258d18
date added to LUP
2023-12-19 10:23:26
date last changed
2024-04-24 14:13:32
@article{5a30b44a-386b-4580-985e-116870258d18,
  abstract     = {{Background - Correct blood group typing is the cornerstone of transfusion medicine. In patients whose gastrointestinal wall is compromised, bacterial enzymes can cause deacetylation of blood group A, converting N-acetylgalactosamine to galactosamine, which resembles the B-defining galactose. This acquired B (acqB) phenomenon, first described 1959, can lead to blood grouping errors. Herein, we explore the possibility of producing acqB red blood cells (RBCs) by enzymatic conversion, for quality control purposes.<br/>Materials and methods - A1 RBCs along with group B and O control RBCs were subjected to enzymatic digestion by bacterially derived deacetylase, FpGalNAcDeAc. RBCs were tested with monoclonal anti-A, anti-B, acquired-B-reactive anti-B (ES4 clone), and a panel of donor plasmas. Standard serological techniques were used. RBCs were subsequently frozen by a glycerolisation method, stored at −80 oC, and thawed for testing to evaluate stability.<br/>Results - Specific deacetylation of A antigen was achieved at both enzyme concentrations. Enzymatically modified A1 RBCs reacted 1-3+ with clone ES4 but not with other anti-B reagents. Group B and O RBC controls reacted as expected. Crossmatch testing revealed 1-3+ reactions in 17/26 group A plasma with modified RBCs but not with untreated controls. Furthermore, 2/3 group AB plasmas reacted strongly with the modified RBCs. The acqB phenotype was maintained upon freezing/thawing.<br/>Discussion - Using a recombinant deacetylase, we demonstrated the feasibility of creating acqB RBCs from donor A1 RBCs, thus providing a reagent for quality assurance of monoclonal anti-B. These RBCs can be frozen once treated, providing a reliable source of this unusual but clinically important phenotype.}},
  author       = {{Hagman, Jennifer Ricci and Rahfeld, Peter and Withers, Stephen G and Kizhakkedathu, Jayachandran N and Olsson, Martin L and Storry, Jill R}},
  issn         = {{1723-2007}},
  language     = {{eng}},
  month        = {{09}},
  publisher    = {{Simiti Servizi SRL}},
  series       = {{Blood Transfusion}},
  title        = {{Use of a recombinant deacetylase to convert A1 red blood cells to the acquired B phenotype for quality control purposes}},
  url          = {{http://dx.doi.org/10.2450/BloodTransfus.584}},
  doi          = {{10.2450/BloodTransfus.584}},
  year         = {{2023}},
}