Antibodies to the protein core of the small cell lung cancer workshop antigen cluster-w4 and to the leucocyte workshop antigen CD24 recognize the same short protein sequence leucine-alanine-proline.
We recently described the identity of the small cell lung cancer (SCLC) cluster-w4 antigen and the human B cell differentiation marker CD24, a glycosylphosphatidylinositol (GPI)-anchored, highly glycosylated surface molecule of only 31-35 amino acids [15]. The specificities of three anti-cluster-w4 and of eleven anti-CD24 MoAbs have been investigated with respect to their binding capacity to the protein core of cluster-w4/CD24 antigen. Four overlapping peptides spanning this protein core were synthesized. MoAbs shown to bind to two overlapping peptides by antibody binding inhibition using the cluster-w4/CD24-positive SCLC cell line SW2 and by direct peptide binding detected in an ELISA were investigated in more detail. To determine the exact epitopes recognized by these MoAbs, an epitope mapping assay using peptides synthesized onto polyethylene pins was established. The three anti-cluster-w4 MoAbs SWA11, SWA21 and SWA22 and the anti-CD24 MoAbs OKB2 and ALB9 recognized the same short leucine-alanine-proline (LAP) sequence in an area without potential glycosylation sites close to the GPI anchor of the protein core of the cluster-w4/CD24 antigen.
Weber
E
E
Department of Medicine, University Hospital, Zurich, Switzerland.
Lehmann
H P
HP
Beck-Sickinger
A G
AG
Wawrzynczak
E J
EJ
Waibel
R
R
Folkers
G
G
Stahel
R A
RA
D000595:Amino Acid Sequence; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000912:Antibodies, Neoplasm / Q000276:immunology; D015703:Antigens, CD / Q000276:immunology*; D000943:Antigens, Differentiation / Q000276:immunology; D000951:Antigens, Neoplasm / Q000276:immunology*; D051927:CD24 Antigen; D018288:Carcinoma, Small Cell / Q000276:immunology*; D000939:Epitopes / Q000032:analysis*; D006801:Humans; D008175:Lung Neoplasms / Q000276:immunology*; D008562:Membrane Glycoproteins; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000276:immunology*
D000911:Antibodies, Monoclonal; D000912:Antibodies, Neoplasm; D015703:Antigens, CD; D000943:Antigens, Differentiation; D000951:Antigens, Neoplasm; D051927:CD24 Antigen; C497889:CD24 protein, human; D000939:Epitopes; D008562:Membrane Glycoproteins; D010446:Peptide Fragments