Molecular identity of a pan cancer marker, CA215.
The molecular nature of cancer-associated antigen, CA215 which reacts with RP215 monoclonal antibody and its unique epitope(s)was characterized. RP215 was initially selected and produced from one of 3,000 hybridomas which were generated from mice immunized with the cell extract of OC-3-VGH ovarian cancer cells. This cancer-associated antigen from various sources including cancer cell extract, shed culture medium and affinity-purified forms was analyzed by MALDI-TOF MS (Matrix Adsorption Laser Desorption Ionization-Time of Flight Mass Spectrometry), Western blot, carbohydrate profiling as well as enzyme immunoassays. The results of this study showed that CA215 is homologous to the heavy chains of human immunoglobulins with molecular sizes ranging from 50 to 70 KDa, when probed with RP215 or anti-human immunoglobulin G, A or M. Treatments of cancer cells with NaIO(4) drastically reduce RP215 binding to the carbohydrate-associated epitope(s) of CA215 located on the variable domain of the human immunoglobulin heavy chains. Further studies indicated that CA215 is predominantly expressed by cancer cells in both secreted and membrane-bound monomeric forms. The carbohydrate-associated epitope(s) with pH-sensitive immunoactivity appear to be present only in cancer cell-derived immunoglobulins, but not in normal human immunoglobulins. Compared to normal immunoglobulin G, CA215 contains a significantly higher percentage of N-acetyl and N-glycoyl neuraminic acid (28% vs. 8%) in the O-linked glycans, but a lower content of N-acetylglucosamine (28% vs. 41%) in the N-linked ones. It was concluded from this study that RP215 reacts specifically with carbohydrate-associated epitope(s) of immunoglobulin heavy chains expressed by various human cancer cells.
Lee
Gregory
G
Andrology Laboratory, Department of Obstetrics and Gynaecology, The University of British Columbia, Vancouver, Canada. andr@interchange.ubc.ca
Laflamme
Emily
E
Chien
Chin-Hsiang
CH
Ting
Hong Hoi
HH
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal; D000951:Antigens, Neoplasm / Q000032:analysis / Q000737:chemistry / Q000235:genetics*; D003429:Cross Reactions; D015180:Electrophoresis, Gel, Two-Dimensional; D000939:Epitopes / Q000032:analysis; D005260:Female; D007118:Immunoassay; D007143:Immunoglobulin Heavy Chains / Q000032:analysis / Q000737:chemistry; D051379:Mice; D008969:Molecular Sequence Data; D009369:Neoplasms / Q000175:diagnosis*; D010051:Ovarian Neoplasms / Q000276:immunology*; D010446:Peptide Fragments / Q000032:analysis / Q000737:chemistry; D011247:Pregnancy; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
D000911:Antibodies, Monoclonal; D000951:Antigens, Neoplasm; C541379:CA215 antigen, human; D000939:Epitopes; D007143:Immunoglobulin Heavy Chains; D010446:Peptide Fragments