Paper-based assay for red blood cell antigen typing by the indirect antiglobulin test

Abstract

A rapid and simple paper-based elution assay for red blood cell antigen typing by the indirect antiglobulin test (IAT) was established. This allows to type blood using IgG antibodies for the important blood groups in which IgM antibodies do not exist. Red blood cells incubated with IgG anti-D were washed with saline and spotted onto the paper assay pre-treated with anti-IgG. The blood spot was eluted with an elution buffer solution in a chromatography tank. Positive samples were identified by the agglutinated and fixed red blood cells on the original spotting area, while red blood cells from negative samples completely eluted away from the spot of origin. Optimum concentrations for both anti-IgG and anti-D were identified to eliminate the washing step after the incubation phase. Based on the no-washing procedure, the critical variables were investigated to establish the optimal conditions for the paper-based assay. Two hundred ten donor blood samples were tested in optimal conditions for the paper test with anti-D and anti-Kell. Positive and negative samples were clearly distinguished. This assay opens up new applications of the IAT on paper including antibody detection and blood donor-recipient crossmatching and extends its uses into non-blood typing applications with IgG antibody-based diagnostics. Graphical abstract A rapid and simple paper-based assay for red blood cell antigen typing by the indirect antiglobulin test.

Indirect antiglobulin paper test for red blood cell antigen typing by flow-through method

“…However, IgG immunoglobulin antibodies (IgG) have performed very poorly. 12 This simple paper test validates the concept of agglutinating IgG-sensitised RBCs on paper. RBC antigen typing with IgG antibodies is essential as some critical blood group antibodies are rarely IgM antibodies, such as for anti-Duffy antibodies (anti-Fy a ).…”

“…12 Removing the washing step requires a precisely controlled ratio between the quantity of antibody, antigen on RBC and anti-IgG. 12 Removing the washing step requires a precisely controlled ratio between the quantity of antibody, antigen on RBC and anti-IgG.…”

“…4,7,9,12 Li et al 7 underlined the signicance of pH of the ushing buffer at about 7 as the positive charges can enhance RBC and antibody interactions. 4,7,9,12 Li et al 7 underlined the signicance of pH of the ushing buffer at about 7 as the positive charges can enhance RBC and antibody interactions.…”

“…Test papers and RBC samples were prepared based on the no-wash method reported by Yeow et al 12 Firstly, 10 mL of anti-IgG FFMU in a 1/4 dilution (25% v/v in PBS) was spotted on 2 cm  2 cm Kleenex paper towel and le to dry in laboratory conditions for 40 minutes (min). Test papers and RBC samples were prepared based on the no-wash method reported by Yeow et al 12 Firstly, 10 mL of anti-IgG FFMU in a 1/4 dilution (25% v/v in PBS) was spotted on 2 cm  2 cm Kleenex paper towel and le to dry in laboratory conditions for 40 minutes (min).…”

Paper and Fiber-Based Bio-Diagnostic Platforms: Current Challenges and Future Needs

“…It requires specifically tailored physical properties of the paper/fiber that can highly encourage analyte-surface interaction through a multitude of forces. In spite of the significant advances in fabrication strategies, paper-based platforms suffer from a lack of control over their physical properties [4,77]. Paper segments in most of the developed platforms are selected from commercially available products (Table 1) possessing certain physical characteristics, including capillary flow rate, surface area, porosity, permeability, and wettability.…”