1998 Volume 10 Issue 55 Pages 393-403
When an affinity ligand is linked to a soluble ionic matrix, the conjugate can specifically change the electrophoretic mobility of a protein which has affinity for the ligand. We refer to the conjugate as an “affinophore” and the electrophoresis using the affinophore “affinophoresis”. Succinylated glutathione is useful as a matrix for monoliganded affinophores.
Iodoacetylated p-aminophenyl glycosides can be coupled to the thiol of the matrix to form affinophores for lectins. The affinophore having a mannoside as a ligand induced mobility change of pea lectin. The analysis of the competitive inhibition of the affinophoresis by neutral sugars using capillary electro-phoresis allowed the determination of the affinity constants for these sugars with very small amounts of a lectin sample and with high precision. The principle of the analysis, preparation of the affinophore and salient points of the experimental procedure ares described.
