Micro-sequencing strategies for the human A33 antigen, a novel surface glycoprotein of human gastrointestinal epithelium Journal Article


Authors: Moritz, R. L.; Ritter, G.; Catimel, B.; Cohen, L. S.; Welt, S.; Old, L. J.; Burgess, A. W.; Nice, E. C.; Simpson, R. J.
Article Title: Micro-sequencing strategies for the human A33 antigen, a novel surface glycoprotein of human gastrointestinal epithelium
Abstract: Monoclonal antibody (mAb) A33, which recognizes a M-r similar to 43000 differentiation antigen (A33) expressed in normal human colonic and small bowel epithelium as well as in 95% of colon cancers, shows specific targeting of colon cancer in humans and is currently being evaluated for clinical use. Here, we describe strategies for the purification and structural analysis of the A33 antigen from the human colorectal carcinoma cell lines LIM1215 and SW1222. Edman degradation of the intact protein and nine peptides, derived by proteolytic digestion of the A33 antigen with Asp-N endoproteinase, thermolysin, trypsin and pepsin followed by micropreparative reversed-phase high-performance liquid chromatography, allowed the unambiguous sequence assignment of 153 amino acid residues; these data reveal one N-glycosylation sequeon in Asp-N endoproteinase peptide D4, and a disulfide linkage between peptides D1 and D4. This amino acid sequence information has facilitated the cloning and subsequent sequencing of a cDNA for the A33 antigen which demonstrates that it is a novel human cell surface molecule of the immunoglobulin superfamily. (C) 1998 Elsevier Science B.V.
Keywords: proteins; antigens; peptides; glycoproteins; advanced; colon-cancer; cell-line; purification; phase i/ii; performance liquid-chromatography; monoclonal-antibody a33; biosensor; columns
Journal Title: Journal of Chromatography A
Volume: 798
Issue: 1-2
ISSN: 0021-9673
Publisher: Elsevier B.V.  
Date Published: 1998-03-06
Start Page: 91
End Page: 101
Language: English
ACCESSION: WOS:000072695800011
DOI: 10.1016/s0021-9673(97)01031-5
PROVIDER: wos
PUBMED: 9542130
Notes: Article; Proceedings Paper -- Presented at 21st International Symposium on High Performance Liquid Phase Separations and Related Techniques, which took place 1997 Jun 22-27 in Burmingham, England -- Source: Wos
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MSK Authors
  1. Leonard Cohen
    18 Cohen
  2. Gerd Ritter
    158 Ritter
  3. Lloyd J Old
    386 Old
  4. Sydney   Welt
    57 Welt