A newly developed gel centrifugation test for quantification of RBC‐bound IgG antibodies and their subclasses IgG1 and IgG3: comparison with flow cytometry

Lynen, R.; Krone, Olaf; Legler, Tobias J.; Köhler, Michael; Mayr, Wolfgang R.

doi:10.1046/j.1537-2995.2002.00076.x

Cited by 27 publications

(32 citation statements)

References 15 publications

(28 reference statements)

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“…It has been seen that in so called “DAT negative AIHA”, more sensitive techniques such as enzyme linked DAT, flow cytometry (FC) and gel cards can detect IgG or C3d molecules coating the red cells. [89]…”

Section: Introductionmentioning

confidence: 99%

Autoimmune hemolytic anemia: From lab to bedside

Chaudhary¹,

Das²

2014

Asian J Transfus Sci

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Autoimmune hemolytic anemia (AIHA) is not an uncommon clinical disorder and requires advanced, efficient immunohematological and transfusion support. Many AIHA patients have underlying disorder and therefore, it is incumbent upon the clinician to investigate these patients in detail, as the underlying condition can be of a serious nature such as lymphoproliferative disorder or connective tissue disorder. Despite advances in transfusion medicine, simple immunohematological test such as direct antiglobulin test (DAT) still remains the diagnostic hallmark of AIHA. The sensitive gel technology has enabled the immunohematologist not only to diagnose serologically such patients, but also to characterize red cell bound autoantibodies with regard to their class, subclass and titer in a rapid and simplified way. Detailed characterization of autoantibodies is important, as there is a relationship between in vivo hemolysis and strength of DAT; red cell bound multiple immunoglobulins, immunoglobulin G subclass and titer. Transfusing AIHA patient is a challenge to the immunohematologist as it is encountered with difficulties in ABO grouping and cross matching requiring specialized serological tests such as alloadsorption or autoadsorption. At times, it may be almost impossible to find a fully matched unit to transfuse these patients. However, transfusion should not be withheld in a critically ill patient even in the absence of compatible blood. The “best match” or “least incompatible units” can be transfused to such patients under close supervision without any serious side-effects. All blood banks should have the facilities to perform the necessary investigations required to issue “best match” packed red blood cells in AIHA. Specialized techniques such as elution and adsorption, which at times are helpful in enhancing blood safety in AIHA should be established in all transfusion services.

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Section: Introductionmentioning

confidence: 99%

Autoimmune hemolytic anemia: From lab to bedside

Chaudhary¹,

Das²

2014

Asian J Transfus Sci

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“…This difference in results between the two studies may have been due to the increased number of samples in the latter study. Lynen et al [6] found a significant difference in the amount of RBC-bound IgG1 when studying 12 patients with and without severe IHA, but later did not find a significant difference when studying 65 patients with and without IHA [7].…”

Section: Detection and Quantitation Of Globulins Red Blood Cell-boundmentioning

confidence: 98%

“…There is a number of methods used to determine the subclass of IgG antibodies, and FC has been shown to be useful [6,7,15,16]. Garratty et al [16] showed that 12 antibodies that could not be subclassed using a routine capillary method [17] (i.e., all four subclass antisera were nonreactive with the antibody-coated RBCs) could be subclassed using FC.…”

Section: Detection and Quantitation Of Globulins Red Blood Cell-boundmentioning

confidence: 99%

“…Thus FC appeared to be a more sensitive method in some cases. Lynen et al [7] compared subclassing by the gel centrifugation test with FC and found that the two methods were comparable when testing the more strongly sensitized RBCs, but FC was better at obtaining results when testing the more weakly sensitized RBCs. FC has also been used in the detection of RBC-bound C3, IgM and IgA [18][19][20][21][22].…”

Section: Detection and Quantitation Of Globulins Red Blood Cell-boundmentioning

confidence: 99%

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Flow Cytofluorometric Analysis in Red Blood Cell Immunology

Arndt¹,

Garratty²

2004

Transfus Med Hemother

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Flow cytofluorometry (FC) is primarily used in the clinical laboratory for the analysis of white blood cells but also has applications in the study of red blood cell (RBC) immunology. When studying RBCs by FC it is important to avoid agglutination of the RBCs, which ironically is the endpoint for most immunohematologic assays. When studying low levels of RBC-bound antibody or RBC antigens with low copy number, background noise due to autofluorescence and nonspecific binding need to be taken into account. It is also important to ensure that reagents (e.g. antisera) used by FC have been standardized for use by FC. Applications of FC in RBC immunology include i) detection and quantitation of globulin (RBC-bound or free in serum), ii) detection and quantitation of RBC antigens, and iii) detection and quantitation of mixed cell populations. FC can be used to differentiate the amount of IgG on strongly IgG-coated RBCs, as well as to detect small amounts of RBC-bound IgG. FC has been used to quantitate antibody that is free in serum, e.g. anti-D in a pregnant woman or in immune globulin preparations. Many RBC antigens have been studied by FC to determine the number of antigen sites, the density distribution, and phenotype-related differences (e.g. zygosity). FC has been found to be very useful for studying mixed cell populations; applications include fetomaternal hemorrhage quantitation, determination of survival/ clearance of transfused RBCs, phenotyping of transfused patients’ RBCs, evaluation of bone marrow transplant engraftment, and study of RBC chimerism and mosaicism.

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“…Flow cytometry (FC) has been previously utilized for antibody detection, including the detection of red cell autoantibodies (4,5), HLA alloantibodies (6,7), platelet crossmatching (8)(9)(10), and other immmunohematological applications (11,12). Recently, FC was also introduced for the screening of antibodies to red cells for automation by Roback et al (3,13).…”

Section: Introductionmentioning

confidence: 99%