Multiplex blood group typing by cellular surface plasmon resonance imaging

Abstract

BACKGROUND: Blood-group typing of donors and patients is essential to avoid incompatible transfusions. Transfusion of incompatible RBCs may result in alloimmunization complicating future transfusions or in the presence of antibodies in adverse reactions. With more than 300 blood group antigens identified, it is difficult to provide fully compatible blood. Currently, standard practice is to match for the most immunogenic antigens. While the current agglutination-based RBC-typing methods are reliable for testing a selected number of antigens, they are not easily adaptable for high-throughput multiplex blood typing beyond the current standard.STUDY DESIGN AND METHODS: Surface plasmon resonance (SPR) is a label-free method to follow molecular-and, very recently, also cellular-interactions in real time. Demonstration of binding of RBCs to blood group antigen-specific antibodies by SPR has already been achieved. Here, we demonstrate the generation of an SPR array equipped with clinically relevant blood group antibodies (A, B, and Rh blood groups). To validate this method, we blindly compared typing of 946 blood donors with results of current diagnostic agglutination-based methods.RESULTS: RBC typing was achieved by monitoring RBC binding to blood group-specific antibodies on the sensor simultaneously within 5 minutes per sample. Regeneration of the chip was robust, allowing for typing of at least 100 samples. The typing results gave a 100% match with classical serology with all antibodies tested besides anti-E/e monoclonals, which gave inconsistent results due to low antibody specificity.CONCLUSION: This study demonstrates that SPRbased RBC typing for multiple antigens can be realized simultaneously with high-quality antibodies, enabling reduced hands-on time and possibly improving cost efficiency.I ncompatible blood transfusion can lead to transfusion reactions or to immunization of the recipient, who generates antibodies that can lead to serious complications in subsequent transfusions. In the case of preexisting alloantibodies, blood transfusion can therefore be given only with RBCs negative for the corresponding blood group antigen. To date, more than 300 blood group antigens have been identified in 36 blood group systems, 1 with varying clinical significance. 2,3 Typing is standardly provided for only limited blood groups with naturally occurring anti-A and B antibodies in antigen-negative individuals, and for the highly immunogenic D antigen. These are universally accepted as important steps in pretransfusion testing. 4 Limited phenotype-matched blood transfusion (including C, ABBREVIATIONS: cSPRi = cellular surface plasmon resonance imaging; S = sedimentation response value; SPR = surface plasmon resonance; SPRi = surface plasmon resonance imaging; T = total response. From the c, E, e, and K antigens) is also recommended in some countries for women during and before childbearing years, to prevent formation of antibodies potentially causing hemolytic disease of the fetus and newborn. 5 Extended phenotype-matched ...

Blood Group Testing

“…In some applications, the sensitivity and versatility of the label-free, real-time technique outperform traditional methods like enzyme-linked immunosorbent assays or fluorescence (70). Generally, RBC antigen-specific IgM or IgG antibodies immobilized on the surface of SPR can be used to identify blood groups (71,72). The intensity of agglutination depends on RBC adhesion and wall shear stress (73).…”

FcγR Binding and ADCC Activity of Human IgG Allotypes

“…To further investigate the strong functional difference between L291-and P291-expression variants despite lack of noticeable affinity differences for FcγRIIIa, we used a cellular SPR setting which measures avidity rather than affinity (Figure 6A) (54)(55)(56). Small differences in affinity may not easily be measured directly, but may result in observable larger differences in avidity.…”

Flipped Quick-Response Code Enables Reliable Blood Grouping

“…Paper-based colorimetric assays, featuring rapidity and convenience, hold great promise for point-of-care (POC) blood typing with naked-eye readouts. − However, these approaches frequently encounter inevitable environmental interference and low precision during the process of readout. , Moreover, the simultaneously grouping of multiplexed blood types such as ABO and Rhesus (Rh) always depends on high-throughput signal transduction strategies and highly cost-effective readout methods. − Our group previously invented a dye-assisted grouping methodology for rapid blood typing by identifying blood component-induced color changes after staining these components using the dye bromocresol green (BCG); we also explored a disk-based approach for multiplexed blood grouping within 2 min by using machine learning-empowered spectrometry . However, the latter still faces difficulties in dealing with field-deployable samples because spectrometers with an integrating sphere represent a heavy burden to guarantee the stability and repeatability of a result when measuring the resultant reflectance spectra, which significantly lowers the on-site feasibility.…”