Structural probing of human lutropin using antibodies raised against synthetic peptides constructed by classical and multiple antigen peptide system approaches.
Antibodies were elicited against a synthetic peptide which encompassed two different regions of the human lutropin beta-subunit (hLH-beta). These antibodies were raised against either the peptide which was assembled using a conventional approach and conjugated to the tetanus toxoid, or with the peptide assembled using the multiple antigen peptide system approach. Automated simultaneous synthesis of the two forms of the immunizing peptide was successfully achieved. Animal injected with the peptide conjugated to tetanus toxoid produced high titers of antibodies to the synthetic peptide, but did not bind to the native hLH-beta subunit. In contrast, antisera induced by the peptide in its MAP form displayed reactivity with both the peptide and the native hLH-beta subunit; these latter antisera appeared to preferentially recognize the beta 47-55 portion of the molecule and were able to bind to the beta-subunit of human choriogonadotropin. Present results demonstrate that the beta 47-55 region is accessible to antibody binding and appears to be located at the surface of both hLH-beta and hLH. Moreover, this study confirms that the MAP approach provides a chemically unambiguous method for obtaining antibodies of predetermined specificity, capable of recognizing cognate sequences of various native proteins.
Unité d'Immunochimie, Institut Gustave Roussy, Villejuif, France.
F
F
Troalen
A
A
Razafindratsita
A
A
Puisieux
T
T
Voeltzel
C
C
Bohuon
D
D
Bellet
J M
JM
Bidart
D000595:Amino Acid Sequence; D000918:Antibody Specificity / Q000276:immunology; D002850:Chromatography, Gel; D000939:Epitopes / Q000032:analysis; D006801:Humans; D007106:Immune Sera / Q000096:biosynthesis; D007986:Luteinizing Hormone / Q000276:immunology*; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000138:chemical synthesis / Q000276:immunology*; D013329:Structure-Activity Relationship
D000939:Epitopes; D007106:Immune Sera; D010446:Peptide Fragments; D007986:Luteinizing Hormone