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Article | 15-February-2021

ZZAP treatment of red blood cells

Principle ZZAP is a mixture of a sulfhydryl reagent (dithiothreitol [DTT]) and a proteolytic enzyme (papain or ficin). It was first described by Branch and Petz in their 1982 article in the American Journal of Clinical Pathology.1 ZZAP is used to dissociate IgG and complement from red blood cells (RBCs), an action that neither reagent can achieve alone. According to Branch and Petz, it is believed that ZZAP “reduces interchain disulfide linkages, increasing exposure of the IgG polypeptides to

S.I. Marckwardt

Immunohematology, Volume 35 , ISSUE 1, 9–10

Article | 14-October-2020

Low-incidence MNS antigens associated with single amino acid changes and their susceptibility to enzyme treatment

MNS antigens are carried on glycophorin A (GPA), glycophorin B (GPB), or their variants. Antigens at the N-terminus of GPA are sensitive to cleavage by ficin, papain, and trypsin but are resistant to α-chymotrypsin. Antigens at the N-terminus of GPB are sensitive to cleavage by ficin, papain, and α-chymotrypsin but are resistant to trypsin treatment. These characteristics have been used to aid in the identification of blood group alloantibodies. Recent molecular analyses have

Marion E. Reid, Jill Storry

Immunohematology, Volume 17 , ISSUE 3, 76–81

Case report | 01-December-2019

A case of masquerading alloantibodies:  the value of a multitechnique approach

, D+ blood type showing strong reactivity with all cells tested  in the forward and reverse ABO, in the D testing as well as in a three-cell antibody screen. The initial assumption was that the plasma contained a cold autoantibody. Subsequent testing, including the use of gel column technology, ficin-treated cells, and antisera for phenotyping, showed the apparent cold autoantibody to be a red herring. Additional tube testing at immediate spin, 37°C, and indirect antiglobulin test (IAT

Paula M.S. Wennersten, Laurie J. Sutor

Immunohematology, Volume 30 , ISSUE 3, 117–120

Article | 27-April-2020

On a much higher than reported incidence of anti-c in R1R1 patients with anti-E

solution for tube IATs (15 min at 37°C,anti-IgG),ficin-tube IATs (30 min at 37°C,anti-IgG + anti-C3), and gel IATs (untreated or ficin-treated RBCs or both, anti-IgG gels). LISS-tube or gel IATs with untreated RBCs revealed anti-c in 32 patients with anti-E. When gel-IAT and ficin-pretreated RBCs were used, 21 additional patients with anti-E were found to have anti-c. In samples from 26 R1R1 patients with anti-E,anti-c was not demonstrable by ficin-gel IATs,and in 3 cases,the ficin-gel tests

W. John Judd, Louann R. Dake, Robertson D. Davenport

Immunohematology, Volume 21 , ISSUE 3, 94–96

Review | 01-December-2019

Warm autoadsorption with enzyme-treated red blood cells

Farai Tsimba-Chitsva, Susanne Bishop, Kelly Kezeor

Immunohematology, Volume 28 , ISSUE 3, 88–90

Article | 06-December-2020

Anti-Uz found in mother's serum and child's eluate

A saline-reactive antibody, anti-Uz, that reacted stronger with S+ than with S- red blood cells (RBCs) and failed to react with U- or ficin-treated RBCs has been previously reported. We describe an antibody of similar specificity in the postpartum serum of an untransfused woman and the eluate from her fourth child's cord RBCs. The mother's RBCs typed S-s+U+, He+(weak), and appeared to have normal glycophorin A and B content, as determined by immunoblotting. The direct antiglobulin test

Sandra M. Read, Mary M. Taylor, Marion E. Reid, Mark A. Popovsky

Immunohematology, Volume 9 , ISSUE 2, 47–49

Article | 03-November-2020

The gel test: use in the identification of unexpected antibodies to blood group antigens

The IgG GEL test was compared with the LISS tube test (Löw and Messeter’s low-ionic-strength saline) for antibody identification. The suitability of red blood cells (RBCs) pretreated with ficin, dithiothreitol (DTT), or chloroquine diphosphate (CDP) also was assessed for use in the GEL test. In addition, time-in-motion studies were performed comparing GEL (12 panels per batch) with polyethylene glycol (PEG) tube tests (3 panels per batch). In 57 antibody identification studies, there

W. John Judd, E.Ann Steiner, Pamela C. Knaf, Colleen Masters

Immunohematology, Volume 14 , ISSUE 2, 59–62

Article | 16-November-2020

A second example of anti-Esa, an antibody to a high-incidence Cromer antigen

A blood sample contained an antibody to a high-incidence antigen that reacted with all red blood cells (RBCs) tested by the indirect antiglobulin test (IAT). The antibody reacted with papain-, ficin-, and trypsin-treated RBCs, but not with α-chymotrypsin-treated RBCs. This pattern of reactivity suggested the possibility that the antibody was recognizing an antigen in the Cromer blood group system. Tests against RBCs deficient in decay-accelerating factor (which carries the Cromer antigens

Marion E. Reid, Roselyn Marfoe, Anita Mueller, Patricia A. Arndt, Laima Sausais, Peggy Spruell

Immunohematology, Volume 12 , ISSUE 3, 112–114

Article | 30-November-2020

First example of Rh:-32,-46 red cell phenotype

The red cells of a white male blood donor typed as Rh:-1, -2, -3, w4, w5, 6, -17, w19, -31, -32, -34, and -46. Although the donor has no history of transfusion, his serum contains an alloantibody that is weakly reactive with most red blood cells (RBCs) tested. Only Rhnull and D-- RBCs are nonreactive. Reactivity is enhanced with ficin- or papain-treated RBCs and is unaffected by AET or DTT treatment of the RBCs. Previously described Rh:-46 RBCs have been of deletion types D--, D•&bull

Jill Storry, Michael Gorman, Nancy I. Maddox, Ella Toy, Peter D. Issitt, Delores M. Mallory

Immunohematology, Volume 10 , ISSUE 4, 130–133

Report | 14-March-2020

Allogeneic adsorptions: a comparison of the traditional method with a modified PEG adsorption method

The purpose of this study is to demonstrate the benefits of enhancing adsorptions with PEG. Allogeneic adsorptions were performed on 20 patient samples containing warm reactive autoantibodies with two volumes of adsorbing RBCs; results using unenhanced adsorptions were compared with those using PEG-enhanced adsorptions and with using untreated adsorbing RBCs and ficin-treated adsorbing RBCs. Two volumes of adsorbing RBCs, one volume of serum, and one volume of PEG were used. The number of

Mandy E. Etem, Barbara Laird-Fryer, Marie P. Holub, John J. Hedl, Daniel B. Symington, Dolores Figueroa

Immunohematology, Volume 26 , ISSUE 3, 104–108

Article | 03-November-2020

Warm autoimmune hemolytic anemia associated with an IgM autoanti-Ge

nonreactive. An acid eluate was only weakly reactive, but a 56°C heat eluate strongly agglutinated untreated RBCs (3+). Ficin-treated RBCs were nonreactive. En(a−) RBCs were strongly reactive, but Ge− RBCs were nonreactive. The anti-Ge in the eluate was IgM. The patient’s untreated RBCs were shown, by flow cytometry, to be weakly Ge+. This is the first report of IgM-mediated warm AIHA associated with autoanti-Ge.

Thom S. Sererat, Douglas W. Veidt, Patricia A. Arndt, George Garratty

Immunohematology, Volume 14 , ISSUE 1, 26–29

Article | 20-December-2020

Two cases of autoantibodies that demonstrate mimicking specificity in the Duffy blood group system

(3+), Fy:1,2 (3+) RBCs at the antiglobulin phase (AGT) but were nonreactive with Fy: -1,- 2 RBCs. The anti­bodies in both cases were nonreactive with papain- or ficin-treated RBCs and reacted with Rhnull RBCs, which ruled out anti-Fy3 and anti-Fy5, respectively. These cases suggest mimicking autoantibody specificities in the Duffy system. One case may indicate that the degree of reactivity is dependent on the Duffy phenotype of the RBCs tested (Fy: -1,2 > Fy:1,2 > Fy: -1, -2). An

Teresa Y. Harris

Immunohematology, Volume 6 , ISSUE 4, 87–91

Article | 26-October-2020

Anti-Lu9: the finding of the second example after 25 years

:-1,2,6,9 antibody-screening red blood cells (RBCs) using either a low-ionic-saline additive solution or polyethylene glycol for enhancement. Lu:6,9 RBCs were reactive with the serum when ficin- or EDTA/glycine-acid-treated, but nonreactive when trypsin- or α-chymotrypsin-treated. Six known examples of Lu:9 RBCs were reactive with the GR serum. His serum did not contain anti-Lua, anti-HLA-B7 (-Bga) or antibodies to 34 low-incidence antigens tested. We have identified the second example of anti-Lu9

Kayla D. Champagne, Marilyn Moulds, Jo Schmidt

Immunohematology, Volume 15 , ISSUE 3, 113–116

Article | 14-October-2020

Warm autoimmune hemolytic anemia with mimicking anti-c and -E specificities

out. Upon adsorption of the patient’s serum with ficin-treated Cde/Cde RBCs, both antibody specificities were adsorbed; however, the antibodies were not adsorbed with native (untreated) Cde/Cde RBCs. Furthermore, the autoantibody was not adsorbed by Rhnull cells, thereby suggesting Rh specificity. The serum was incompatible with cde/cde RBCs and compatible with Cde/Cde RBCs. The patient was successfully transfused with Cde/Cde RBCs followed by resolution of his anemia, as evidenced by an

Hsin-Yeh Hsieh, Diana L. Moroney, Deanne E. Naumann, D. Jane Hata, Nancy C. Vosnidou, Rovenna L. Kessinger, Nassir Shahab, Nasrollah Hakami, Daniel S. Smith

Immunohematology, Volume 18 , ISSUE 1, 19–22

Article | 14-October-2020

PEG adsorption of autoantibodies causes loss of concomitant alloantibody

Use of polyethylene glycol (PEG) to promote adsorption of autoantibodies is reported to give good recovery of concomitant alloantibodies. In initial experiments, PEG and ZZAP (Ficin and DTT) adsorption procedures were compared for removal of autoantibody and recovery of alloantibody. Postadsorption studies (n = 11) were performed and hemagglutination scores compared. In subsequent studies, equal volumes of alloantibody containing sera, PEG, and antigen-negative red blood cells (RBCs) were used

W. John Judd, Louann Dake

Immunohematology, Volume 17 , ISSUE 3, 82–85

Article | 18-October-2020

Red blood cell diluent composition is important for detection of some anti-E

PEG and/or ficin-treated RBCs only. As a comparison, 25 archived examples of anti-E were detected using RBCs suspended in MTS2 and Ortho 0.8% reagent RBCs. These data show that some anti-E are not detected by Ortho reagent RBCs in MTS Anti-IgG gel cards. However, these anti-E have characteristics of antibodies of questionable clinical significance.

Dania D. Yaskanin, Janice L. Jakway, David J. Ciavarella

Immunohematology, Volume 16 , ISSUE 4, 142–146

Article | 26-October-2020

Naturally-occurring anti-Jka in infant twins

with ficin- or papain-treated RBCs. Monocyte monolayer assays using Jk(a+) RBCs sensitized by either twins' serum were nonreactive (0%). RBCs from both parents typed as Jk(a+b+). Both parents’ antibody detection test results by SPRCA assay were negative. The absence of a history of exposure to allogeneic RBCs or possible passive transfer of maternal or other alloantibody classifies these antibodies as naturally-occurring anti-Jka.

Dawn H. Rumsey, Sandra J. Nance, Mary Rubino, S. Gerald Sandler

Immunohematology, Volume 15 , ISSUE 4, 159–162

Report | 01-December-2019

The production, serologic evaluation, and epitope mapping of ten murine monoclonal Dombrock antibodies

recognized epitopes that were resistant to treatment with ficin, pronase, α-chymotrypsin, and neuraminidase, but sensitive to trypsin and 0.2 M dithiothreitol (DTT). Five have anti-Dob-like specificity. The epitope recognized by MIMA-52 was neuraminidase sensitive, and MIMA-127 epitope recognized a DTT-resistant, linear epitope 90QKNYFRMWQK99 of the Dombrock polypeptide. MIMA-127 was the only one of the ten Dombrock MoAbs mapped to a specific sequence of the Dombrock glycoprotein; the other nine

Magdalena Grodecka, Kazimiera Wasniowska, Gregory Halverson, Karina Yazdanbakhsh, Marion E. Reid, Elwira Lisowska

Immunohematology, Volume 28 , ISSUE 4, 124–129

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