Korean J Transplant.  2023 Sep;37(3):189-196. 10.4285/kjt.23.0033.

The role of acidification of the medium on the erythrocyte agglutinating and adsorbing properties of blood group-specific monoclonal antibodies

• Delevsky YP
• Zinchenko OA

• KMID: 2546485
• DOI: http://doi.org/10.4285/kjt.23.0033

Abstract

Background
Studies on the specificity of ABH antigen-antibody interactions at different pH values are rare. Therefore, the aim of this study was to estimate the effect of acidification of the reacting medium on the agglutinating ability of anti-A monoclonal antibodies (mABs) and their inhibition by glycoconjugates of red blood cell membranes.
Methods
Anti-A mABs were obtained from the Fourth International Workshop on Monoclonal Antibodies Against Human Red Blood Cell and Related Antigens (on July 19-20, 2002, in Paris, France). The glycoconjugates were isolated from erythrocytes' membranes. The inhibition of the lipid and protein isotypes of the blood group A antigen was assessed.
Results
The acidic medium decreased the agglutinating ability of acid immunoglobulin M (IgM) anti-A mABs, in contrast to alkaline immunoglobulin G (IgG) mABs. Meanwhile, at pH 6.5, IgM antibodies exhibited a high adsorption capacity, while IgG antibodies demonstrated a strong adsorption capacity at an alkaline pH. mABs 2-19, 2-28, 2-22, and 2-8 were inhibited by the acidic lipid and protein glycotopes of erythrocyte membranes. However, mAB 2-8 was inhibited by both acidic and alkaline glycotope variants.
Conclusions
The agglutinating and adsorbing abilities of mABs, which are revealed at opposing pH values, should be taken into account during studies of antigen-antibody interactions.

Keyword

Antibody; Adsorption; Erythrocyte; Transferase; Charge

Figure

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  Fig. 1 (A) Anti-A 2-8 monoclonal antibody (mAB) with red blood cells (RBCs) with the blood group A antigen at 4 °C; 4+. (B) Anti-A mAB 2-8 with RBCs with the blood group A antigen at 37 °C; 3+. (C) mAB 2-19 with RBCs with the blood group A antigen at 37 °C at pH 6.0; 2+. (D) Anti-A mAB 2-23 with RBCs with the blood group A antigen at 37 °C; 1+. (E) Adsorbed anti-A mAB 2-8 with RBCs with the blood group A antigen at 37 °C; w+ (3 pt). (F) Adsorbed mAB 2-23 at 37 °C with RBCs with the blood group A antigen; w+ (1 pt) (A–F) ×400. pt, points.

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