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- Halverson, Gregory R, et al.
(författare)
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Murine monoclonal anti-s and other anti-glycophorin B antibodies resulting from immunizations with a GPB.s peptide.
- 2009
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Ingår i: Transfusion. - : Wiley. - 1537-2995 .- 0041-1132. ; 49, s. 485-494
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Tidskriftsartikel (refereegranskat)abstract
- BACKGROUND: The blood group antigens S and s are defined by amino acids Met or Thr at position 29, respectively, on glycophorin B (GPB). Commercial anti-s reagents are expensive to produce because of the scarcity of human anti-s serum. Our aim was to develop hybridoma cell lines that secrete reagent-grade anti-s monoclonal antibodies (MoAbs) to supplement the supply of human anti-s reagents. STUDY DESIGN AND METHODS: Mice were immunized with the GPB(s) peptide sequence TKSTISSQTNGETGQLVHRF. Hybridomas were produced by fusing mouse splenocytes with mouse myeloma cells (X63.Ag8.653). Screening for antibody production was done on microtiter plates by hemagglutination. Characterization of the MoAbs was done by hemagglutination, immunoblotting, and epitope mapping. RESULTS: Eight immunoglobulin G MoAbs were identified. Five antibodies are specific by hemagglutination for s and two MoAbs, when diluted, are anti-S-like, but additional analyses shows a broad range of reactivity for GPB. Typing red blood cells (RBCs) for s from 35 donors was concordant with molecular analyses as were tests on RBCs with a positive direct antiglobulin test (DAT) from 15 patients. The anti-s MoAbs are most reactive with peptides containing the (31)QLVHRF(36) motif, with (29)Thr. By Pepscan analyses, the anti-S-like MoAbs reacted within the same regions as did anti-s, but independently of (29)Met. One antibody was defined serologically as anti-U; however, its epitope was identified as (21)ISSQT(25), a sequence common for both GPA and GPB. CONCLUSION: In addition to their value as typing reagents, these MoAbs can be used to phenotype RBCs with a positive DAT without pre-test chemical modification.
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| 2. |
- Hult, Annika, et al.
(författare)
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Flow cytometry evaluation of red blood cells mimicking naturally occurring ABO subgroups after modification with variable amounts of function-spacer-lipid A and B constructs.
- 2012
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Ingår i: Transfusion. - : Wiley. - 1537-2995 .- 0041-1132. ; 52, s. 247-251
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Tidskriftsartikel (refereegranskat)abstract
- BACKGROUND: Kodecytes bearing synthetic blood group A and B antigens are increasingly being used in transfusion laboratories as serologic mimics of red blood cell (RBC) A(weak) and B(weak) subtypes. The aim of this study was to compare the flow cytometry profile of kodecytes with native ABO subgroups. STUDY DESIGN AND METHODS: A series of A/B kodecytes, each with decreasing A or B antigen expression, were prepared from group O RBCs that were modified with dilutions of function-spacer-lipid KODE technology (FSL) constructs representing a wide serologic range. Using an established flow cytometry method designed for the detection of low levels of A/B antigens, kodecyte profiles were compared with those of native subgroup cells. RESULTS: Kodecytes with positive tube serology from 4+ to 1+ were created with 15 to 2 µg/mL FSL-A or 78 to 10 µg/mL FSL-B transformation solutions. The kodecytes created with higher concentrations of FSL constructs revealed a uniform and/or even distribution of antigens as seen by a single flow cytometry peak more narrow than the broader peaks produced with lower FSL concentrations similar to those found in native A(x) and most B(weak) subgroups. CONCLUSIONS: Although kodecytes are created artificially, they can be designed to mimic the serologic and flow cytometric profiles of native ABO subgroup RBCs.
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