Molecular evolution of specific human antibody against MUC1 mucin results in improved recognition of the antigen on tumor cells.
The MUC1 mucin is differentially expressed and glycosylated in cancer tissue as opposed to healthy tissue. Due to these differences, MUC1 is considered a potential biomarker suitable for cancer diagnosis and therapy. In a previous study, the human MUC1-specific antibody 12ESC-6 was able to bind a sequence variant of the tandem repeat of MUC1 that is not recognized by many other MUC1-specific antibodies. It was also found to bind efficiently to MUC1-carrying cells. We have now used 12ESC-6 as starting point for random mutagenesis to isolate variants with improved ability to bind MUC1 in human tumor tissue. The resulting 12ESC-6 variants were shown to recognize not only the naked MUC1 tandem repeat but even more so glycosylated variants thereof, in particular those carrying the GalNAc (Tn) glycoform. Selected variants of 12ESC-6 demonstrated improved staining of MUC1 on cell lines using flow cytometry and improved staining of the antigen in breast tumor tissue by immunohistochemistry. Molecular evolution and specific fine-tuning thus have the potential to improve the performance of antibody specificities targeting tumor-associated epitopes on MUC1 mucin.
Department of Immunotechnology, Lund University, Lund, Sweden.
Jonas
J
Persson
Malin
M
Bäckström
Henrik
H
Johansson
Karin
K
Jirström
Gunnar C
GC
Hansson
Mats
M
Ohlin
D000595:Amino Acid Sequence; D019943:Amino Acid Substitution; D000818:Animals; D000906:Antibodies / Q000235:genetics*; D000918:Antibody Specificity; D000941:Antigens / Q000032:analysis; D001943:Breast Neoplasms / Q000276:immunology / Q000473:pathology; D016466:CHO Cells; D045744:Cell Line, Tumor; D002850:Chromatography, Gel; D006224:Cricetinae; D003412:Cricetulus; D000939:Epitopes / Q000032:analysis; D019143:Evolution, Molecular; D005260:Female; D006801:Humans; D008297:Male; D008969:Molecular Sequence Data; D018396:Mucin-1 / Q000032:analysis* / Q000235:genetics / Q000276:immunology*; D020411:Oligonucleotide Array Sequence Analysis; D010446:Peptide Fragments / Q000737:chemistry; D011471:Prostatic Neoplasms / Q000276:immunology / Q000473:pathology; D012016:Reference Values; D020449:Repetitive Sequences, Amino Acid; D013601:T-Lymphocytes / Q000276:immunology
D000906:Antibodies; D000941:Antigens; D000939:Epitopes; D018396:Mucin-1; D010446:Peptide Fragments