Debbia Martine, Brossard Yves, Lambin Patrick
Immunology Transfusion Unit, National Institute of Blood Transfusion, Paris, France.
Transfusion. 2005 Jun;45(6):975-83. doi: 10.1111/j.1537-2995.2005.04313.x.
Few data are available on the affinity of maternal anti-D responsible for hemolytic disease of the fetus and the newborn (HDN) and of anti-D used for the prophylaxis of that disease. A method was recently described to measure the affinity (K(a)) of untagged anti-D monoclonal antibodies (MoAbs). In this work, the same method was applied to determine the K(a) of polyclonal anti-D.
O R(1)r red blood cells (RBCs) were sensitized with increasing concentrations of native anti-D in serum samples from immunized mothers and donors and in RhIG preparations. At equilibrium, the amount of anti-D bound to RBCs was measured by enzyme-linked immunosorbent assay. Scatchard and Langmuir equations were used to determine Ka.
The experimental data fitted well with the Scatchard equation (mean r2=0.95) but a better correlation was observed with the Langmuir equation (mean r2=0.99). The mean Ka of anti-D in 11 maternal serum samples, in 6 immunized donors, and in 5 lots of RhIG were 5.6x10(8) per M (from 2.8x10(8) to 12x10(8)/M), 3.9x10(8) per M (from 1.5x10(8) to 6.8x10(8)/M), and 3.4x10(8) per M (from 3.1x10(8) to 4.2x10(8)/M), respectively. The comparison of anti-D affinity in 5 cases of HDN with fetal anemia and in 6 cases of HDN with postnatal anemia showed no significant difference.
The method previously described for anti-D MoAbs was applied to polyclonal anti-D present in the serum of immunized subjects and in immunoglobulin preparations. The experimental data fitted well with the Langmuir equation, and the affinity of polyclonal of anti-D was measured with accuracy.
关于引起胎儿和新生儿溶血病（HDN）的母体抗-D以及用于预防该疾病的抗-D的亲和力，现有数据较少。最近描述了一种测量未标记抗-D单克隆抗体（MoAbs）亲和力（K(a)）的方法。在本研究中，应用相同方法测定多克隆抗-D的K(a)。
用来自免疫母亲和供体的血清样本以及Rh免疫球蛋白（RhIG）制剂中浓度递增的天然抗-D致敏O R(1)r红细胞（RBCs）。在平衡时，通过酶联免疫吸附测定法测量与RBCs结合的抗-D量。使用Scatchard方程和Langmuir方程确定Ka。
实验数据与Scatchard方程拟合良好（平均r2 = 0.95），但与Langmuir方程的相关性更好（平均r2 = 0.99）。11份母体血清样本、6名免疫供体和5批RhIG中抗-D的平均Ka分别为每摩尔5.6×10(8)（范围为2.8×10(8)至12×10(8)/摩尔）、每摩尔3.9×10(8)（范围为1.5×10(8)至6.8×10(8)/摩尔）和每摩尔3.4×10(8)（范围为3.1×10(8)至4.2×10(8)/摩尔）。5例伴有胎儿贫血的HDN和6例伴有产后贫血的HDN中抗-D亲和力的比较显示无显著差异。
先前描述的针对抗-D MoAbs的方法应用于免疫个体血清和免疫球蛋白制剂中的多克隆抗-D。实验数据与Langmuir方程拟合良好，多克隆抗-D的亲和力得以准确测定。
