Malmborg A C, Michaëlsson A, Ohlin M, Jansson B, Borrebaeck C A
Department of Immunotechnology, Lund University, Sweden.
Scand J Immunol. 1992 Jun;35(6):643-50. doi: 10.1111/j.1365-3083.1992.tb02970.x.
Surface plasmon resonance, i.e. detection of changes in refractive index on a surface, was used in a biosensor to evaluate the dissociation/association rate and affinity constants of human monoclonal IgG and IgM antibodies and Fab fragments. The results showed that an observed difference in affinity constants between intact and fragmented IgG anti-tetanus antibody was related to approximately 10-fold differences in dissociation rate constants, since the association rate constants were in the same range, i.e. 2-3 x 10(5) (M-1 s-1). Affinity constants, as determined by conventional solid phase enzyme immunoassay, were substantially higher than the constants produced by the biosensor. Human monoclonal IgM anti-Tn alpha antibodies showed, furthermore, one order of magnitude higher association rate constants, as compared with the IgG antibodies, but since the dissociation rate constants were more than ten times higher, the resulting affinity constants of the anti-carbohydrate IgM antibodies were still somewhat lower than those of the IgG antibodies.
表面等离子体共振，即检测表面折射率的变化，被用于生物传感器中，以评估人单克隆IgG和IgM抗体以及Fab片段的解离/缔合速率和亲和常数。结果表明，完整的和片段化的抗破伤风IgG抗体之间观察到的亲和常数差异与解离速率常数约10倍的差异有关，因为缔合速率常数在相同范围内，即2 - 3×10⁵（M⁻¹ s⁻¹）。通过传统的固相酶免疫测定法测定的亲和常数显著高于生物传感器产生的常数。此外，与人单克隆IgG抗体相比，人单克隆IgM抗Tnα抗体的缔合速率常数高一个数量级，但由于解离速率常数高出十多倍，抗碳水化合物IgM抗体的最终亲和常数仍略低于IgG抗体。
