Factors influencing the reaction of alpha 1-fetoprotein with concanavalin A and Lens culinaris agglutinin in crossed affinoimmunoelectrophoresis.
Concanavalin A (Con A) and lentil lectin (LCA) analysis of alpha-fetoprotein (AFP) glycosylation heterogeneity is used in a variety of clinical situations. We studied the influence of analytical conditions on the separation of AFP glycoforms by using lectin-crossed affinoimmunoelectrophoresis, regardless of the AFP concentration, which can vary over a wide range in biological fluids. We defined the optimal concentration of Con A (2 g/L) and LCA (0.35 g/L) in the first-dimension gel, together with the optimum antigen (AFP)/antibody ratio in the second-dimension gel. The presence of protein in the diluent used for AFP samples was found to change the shape of crossed affinoimmunoelectrophoresis patterns without changing the percentage composition of AFP fractions. The within-run CV was less than 4% for both lectins, and the between-run CV was less than 6.3%. The minimal quantity of AFP that provided a visible pattern with both lectins was 4 ng, corresponding to 50 microL of an 80 micrograms/L AFP sample. These technical conditions allow the cellular origin of AFP to be determined, regardless of the concentration in the sample. Typical AFP lectin patterns of secreting tumors are compared with fetal and cord serum AFP.
Magne
D
D
Laboratoire de Biochimie, Hôpital Necker, Paris, France.
Seta
N
N
Lebrun
D
D
Durand
G
G
Durand
D
D
D000653:Amniotic Fluid / Q000737:chemistry; D006528:Carcinoma, Hepatocellular / Q000097:blood; D003208:Concanavalin A; D005312:Fetal Blood / Q000737:chemistry; D006801:Humans; D007106:Immune Sera; D007123:Immunoelectrophoresis, Two-Dimensional / Q000379:methods* / Q000706:statistics & numerical data; D037102:Lectins; D008113:Liver Neoplasms / Q000097:blood; D037121:Plant Lectins; D013724:Teratoma / Q000097:blood; D000509:alpha-Fetoproteins / Q000032:analysis* / Q000276:immunology
D007106:Immune Sera; D037102:Lectins; D037121:Plant Lectins; D000509:alpha-Fetoproteins; C033619:lentil lectin; D003208:Concanavalin A