title: Quantitative blood group typing using surface plasmon resonance.
abstract: The accurate and reliable typing of blood groups is essential prior to blood transfusion. While current blood typing methods are well established, results are subjective and heavily reliant on analysis by trained personnel. Techniques for quantifying blood group antibody-antigen interactions are also very limited. Many biosensing systems rely on surface plasmon resonance (SPR) detection to quantify biomolecular interactions. While SPR has been widely used for characterizing antibody-antigen interactions, measuring antibody interactions with whole cells is significantly less common. Previous studies utilized SPR for blood group antigen detection, however, showed poor regeneration causing loss of functionality after a single use. In this study, a fully regenerable, multi-functional platform for quantitative blood group typing via SPR detection is achieved by immobilizing anti-human IgG antibody to the sensor surface, which binds to the Fc region of human IgG antibodies. The surface becomes an interchangeable platform capable of quantifying the blood group interactions between red blood cells (RBCs) and IgG antibodies. As with indirect antiglobulin tests (IAT), which use IgG antibodies for detection, IgG antibodies are initially incubated with RBCs. This facilitates binding to the immobilized monolayer and allows for quantitative blood group detection. Using the D-antigen as an example, a clear distinction between positive (>500 RU) and negative (<100 RU) RBCs is achieved using anti-D IgG. Complete regeneration of the anti-human IgG surface is also successful, showing negligible degradation of the surface after more than 100 regenerations. This novel approach is validated with human-sourced whole blood samples to demonstrate an interesting alternative for quantitative blood grouping using SPR analysis.
authors.affiliation: Bioresource Research Institute of Australia (BioPRIA), Australian Pulp and Paper Institute (APPI), Department of Chemical Engineering, Faculty of Engineering, Monash University, Clayton, VIC 3800, Australia.
authors.forename: Whui Lyn
authors.initials: WL
authors.lastname: Then
authors.affiliation: Department of Biochemistry and Molecular Biology, Faculty of Medicine, Nursing and Heath Sciences, Monash University, Clayton, VIC 3800, Australia.
authors.forename: Marie-Isabel
authors.initials: MI
authors.lastname: Aguilar
authors.affiliation: Bioresource Research Institute of Australia (BioPRIA), Australian Pulp and Paper Institute (APPI), Department of Chemical Engineering, Faculty of Engineering, Monash University, Clayton, VIC 3800, Australia. Electronic address: gil.garnier@monash.edu.
authors.forename: Gil
authors.initials: G
authors.lastname: Garnier
mesh_terms: D055910:Antibodies, Immobilized / Q000276:immunology; D001788:Blood Grouping and Crossmatching / Q000379:methods* / Q000706:statistics & numerical data; D004912:Erythrocytes / Q000276:immunology; D006801:Humans; D007141:Immunoglobulin Fc Fragments / Q000276:immunology; D007074:Immunoglobulin G / Q000276:immunology; D012204:Rh-Hr Blood-Group System / Q000276:immunology; D018029:Rho(D) Immune Globulin / Q000276:immunology; D020349:Surface Plasmon Resonance / Q000379:methods* / Q000706:statistics & numerical data
chemical_list: D055910:Antibodies, Immobilized; D007141:Immunoglobulin Fc Fragments; D007074:Immunoglobulin G; C061961:RHO(D) antibody; D012204:Rh-Hr Blood-Group System; D018029:Rho(D) Immune Globulin; C026410:Rho(D) antigen
keywords: Blood group typing; Diagnostic; IgG antibody; Red blood cells (RBCs); Rh phenotype; Surface plasmon resonance