Effect of Ionic Strength on the Serologic Behavior
of Red Cell Isoantibodies

Elliot, Margaret; Bossom, Edith L.; Dupuy, Mary Edith; Masouredis, S.P.

doi:10.1159/000464880

Cited by 5 publications

(6 citation statements)

References 7 publications

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“…We observed the highest antiparallel β‐sheet contents at low salt concentrations. It is well‐known that low ionic strength increases antibody binding . In fact, low ionic strength solutions are routinely used in the red cell laboratory for detection of anti‐red cell antibodies.…”

Section: Discussionmentioning

confidence: 99%

Reactivity of platelet‐activating and nonplatelet‐activating anti‐PF4/heparin antibodies in enzyme immunosorbent assays under different conditions

Nguyen

Wesche

Raschke

et al. 2019

Journal of Thrombosis and Haemostasis

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Background: Enzyme immunosorbent assays (EIA) are widely used to detect human antiplatelet factor 4/heparin antibodies (aPF4/H Abs) to rule out heparin-induced thrombocytopenia. EIAs cannot differentiate between clinically relevant, plateletactivating, and nonrelevant, nonplatelet-activating Abs and only ~50% of patients' sera testing positive by EIA contain antibodies that activate platelets. Recently, we have shown platelet-activating aPF4/H Abs bind more strongly to PF4/H complexes than nonplatelet-activating antibodies. Antigen-antibody interactions are known to depend on electrostatic interactions governed by pH, heat, and ionic strength. We tested whether changes in pH and ionic strength can improve the specificity of EIAs detecting aPF4/H Abs. Methods:We investigated first the conformational change of PF4 when binding to heparin under various pH and salt conditions using circular dichroism spectroscopy, and then the binding of aPF4/H Abs to PF4/H complexes by EIA. Results:Maximal conformational change of PF4 on complexation with heparin was identified at low pH and low salt concentrations. EIA tested with a large number of sera at 50 mmol/L NaCl, pH 6.0 shows a potential to increase the specificity for the detection of platelet-activating aPF4/H Abs. Conclusion:Changing physicochemical conditions may become an approach to better discriminate the signal of platelet-activating and nonactivating PF4/H Abs in antigen tests. K E Y W O R D SaPF4/H antibody, CD spectroscopy, enzyme immunosorbent assays, human PF4, ionic strength, pH

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Section: Discussionmentioning

confidence: 99%

Reactivity of platelet‐activating and nonplatelet‐activating anti‐PF4/heparin antibodies in enzyme immunosorbent assays under different conditions

Nguyen

Wesche

Raschke

et al. 2019

Journal of Thrombosis and Haemostasis

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“…A tally of the blood group antigens listed in the index of Peter D. Issitt's Applied Blood Group Serology published in 1970 12 is a reminder of the impact that reagent red blood cell (RBC) screening and antibody identification had to immunohematology. During this era, the standards of practice were adapted regularly as clinically significant blood group systems were identified and enhancement reagents were introduced, like albumin, 13 low‐ionic‐strength saline (LISS), 14‐16 and hexadimethrine bromide (Polybrene, Sigma‐Aldrich, St Louis, MO) 17,18 . The use of compounds to alter the RBC membrane, like enzymes and ZZAP, also became more common 19‐22 .…”

mentioning

confidence: 99%

Applying molecular immunohematology discoveries to standards of practice in blood banks: now is the time

Denomme

Flegel

2008

Transfusion

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Lessons from more than 100 years of immunohematology exemplify that many critical discoveries were made serendipitously and their more rapid implementation could have benefited transfusion recipients and pregnancies. Constituents of blood that are not essential for the attempted therapeutic benefit of a transfusion are largely removed from today's blood products. We are now moving on to avoid unnecessary exposure to potentially harmful constituents of the therapeutically required cells, like blood group antigens that are foreign to the patient. Cost efficacy needs to be kept in mind but may eventually prove much better than anticipated, once hidden benefits are captured, as we show by examples from past immunohematologic developments. Here, we detail clinical applications for molecular immunohematology advances including "dry-matching" that will improve transfusion outcomes and argue for their widespread implementation by rapid timelines through standards of practice.

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“…Most workers have found that with anti-A and anti-B results are similar with low-ionic-strength medium and with saline (Elliot et al 1964;Löw and Messeter 1974;Moore and Mollison 1976). The advantages of using a low-ionic-strength medium are seen chiefly in tests with IgG antibodies (see section on antiglobulin tests, below).…”

Section: Low Ionic Strengthmentioning

confidence: 89%

“…With many alloantibodies, the titre was found to be increased when red cells were suspended in low-ionic-strength medium but the effect on undiluted serum was not tested (Elliot et al 1964;Hughes-Jones et al 1964b). As discussed above, the uptake of antibody is much more rapid at low ionic strength.…”

Section: The Uptake Of Antibodymentioning

confidence: 99%