Roggenbuck D, König H, Niemann B, Schoenherr G, Jahn S, Porstmann T
Department of Medical Immunology, Medical School (Charité), Humboldt University of Berlin, Germany.
Scand J Immunol. 1994 Jul;40(1):64-70. doi: 10.1111/j.1365-3083.1994.tb03434.x.
Surface plasmon resonance (SPR) was used to investigate the kinetics of interactions between the human monoclonal polyreactive IgM antibody CB03, its Fab as well as its single-chain variable fragment (scFv) and different antigens. From these experiments apparent binding constants were determined and compared with binding constants obtained by ELISA experiments. In SPR studies with the complete antibody, the polyreactivity of the CB03 antibody as derived from ELISA experiments was confirmed. Interaction of scFv with kappa casein and human myoglobin is strong evidence for the location of polyreactivity within the variable domains of the antibody. Apparent binding constants of the complete antibody to immobilized kappa casein (9.2 x 10(7) M-1) and to human myoglobin (1.6 x 10(7) M-1) are up to 83 times higher than those of Fab. The binding constants of the scFv to the above mentioned antigens are again about 10 times lower when compared with Fab, which is mainly due to the lower association rates of the complexes formed by the scFv.
表面等离子体共振（SPR）用于研究人源单克隆多反应性IgM抗体CB03、其Fab片段以及单链可变片段（scFv）与不同抗原之间相互作用的动力学。通过这些实验确定了表观结合常数，并与酶联免疫吸附测定（ELISA）实验获得的结合常数进行了比较。在对完整抗体的SPR研究中，证实了ELISA实验中CB03抗体的多反应性。scFv与κ-酪蛋白和人肌红蛋白的相互作用有力地证明了抗体可变区内多反应性的位置。完整抗体与固定化κ-酪蛋白（9.2×10⁷ M⁻¹）和人肌红蛋白（1.6×10⁷ M⁻¹）的表观结合常数比Fab片段的表观结合常数高出83倍。与Fab片段相比，scFv与上述抗原的结合常数再次降低约10倍，这主要是由于scFv形成的复合物的缔合速率较低。
