What are the types of Rh typing reagents? Describe the preparation and the use of each. a.

Saline reactive reagents Saline reactive reagents, which contain IgM immunoglobulin, were the rst typing reagents available to test for the D antigen. Saline anti-D has the advantage of being low- proteinbased and can be used to test cells that are coated with IgG antibody. The primary disadvantages of saline typing reagents are their limited availability, cost of production, and lengthy incubation time. Because saline anti-D is an IgM immunoglobulin, it cannot be used for weak-D typing.

b. High-protein anti-D reagents High-protein anti-D reagents were developed in the 1940s. Human plasma containing high-titer D-specic antibody is used as the raw material. Potentiators of bovine albumin and macromolecular additives such as dextran or polyvinylpyrrolidone are added to the source material to optimize reactivity in the standard slide and rapid tube tests. These reagents are commonly referred to as high-protein reagents. The presence of potentiators and the higher protein concentration, however, increase the likelihood of false- positive reactions. To assess the validity of the high-protein Rh typing results, a control reagent was manufactured and had to be tested in parallel with each Rh test. If the control reacted, the test result was invalid and had to be repeated using a different technique or reagent anti-D. The major advantages of highprotein anti-D reagents are reduced incubation time and the ability to perform weak-D testing and slide typing with the same reagent. c. Chemically modied IgG anti-D reagents In the late 1970s, scientists chemically modied the IgG anti-D molecule by breaking the disulde bonds that maintain the antibodys rigid shape. This allows the antibody to relax and to span the distance between RBCs in a low-protein medium. The chemically modied reagents can be used for both slide and tube testing and do not require a separate, manufactured Rh control as long as the samples type as A, B, or O. When samples test AB Rh-positive or when the Rh test is performed by itself, a separate saline control must be used to ensure that the observed reactions are true agglutination and not a result of spontaneous agglutination. Fewer false- positive test reactions are obtained because of the lower- protein suspending medium. Because of its lower-protein base and ready availability, the chemically modied anti-D replaced the need for saline anti-D reagents. d. Monoclonal antibody reagents Monoclonal antibody reagents have become available more recently. These reagents are derived from single clones of antibody-producing cells. The antibody-producing cells are hybridized with myeloma cells to increase their reproduction rate and thereby to maximize their antibody-producing capabilities. Because the D antigen appears to be a mosaic and the monoclonal Rh antibodies have a narrow specicity, monoclonal anti-D reagents are usually a combination of mono- clonal anti-D reagents from several different clones to ensure reactivity with a broad spectrum of Rh-positive RBCs. Some companies also blend anti-IgM and anti-IgG anti-D to maximize visualization of reactions at immediate spin testing and to allow indirect antiglobulin testing for weak D with the same reagent. The monoclonal blends can be used for

slide, tube, microwell, and most automated Rh testing. Because these reagents are not humanderived, they lack all potential for transmitting infectious disease. Reference: Harmening, D. (2005). Modern blood banking and transfusion practices (5th Edition). P.142-143. Reporter: TOMITA, Melanie D.