Characterization of protein conjugates using capillary electrophoresis.
With the aim of generating antibodies, a calix[4]arene-crown-6 was coupled to bovine serum albumin. For that purpose, a complete procedure to optimize and characterize the coupling of hydrophobic haptens based on capillary electrophoresis (CE) was developed. We demonstrated the existence of a polynomial relationship between the electrophoretic mobility (mu(ep)) and the hapten density. This correlation was used not only to study the coupling reaction in terms of optimization and kinetics but also to determine the average coupling molar ratio of any given conjugate. An estimation of the heterogeneity of these conjugates by simulation of experimental peaks was also proposed.
Safi
Samir
S
Laboratoire de Chimie Analytique et Minérale, IPHC-DSA (UMR 7178), ULP, CNRS, ECPM, 25 rue Becquerel, F-67087 Strasbourg Cedex, France.
Asfari
Zouhair
Z
Hagège
Agnès
A
D000818:Animals; D002417:Cattle; D019075:Electrophoresis, Capillary / Q000379:methods*; D006241:Haptens / Q000032:analysis / Q000737:chemistry; D008958:Models, Molecular; D008968:Molecular Conformation; D011506:Proteins / Q000032:analysis / Q000737:chemistry*; D012710:Serum Albumin, Bovine / Q000032:analysis / Q000737:chemistry
D006241:Haptens; D011506:Proteins; D012710:Serum Albumin, Bovine