Glycosylation Profiles of the human colorectal cancer A33 antigen naturally expressed in the human colorectal cancer cell line SW1222 and expressed as recombinant protein in different insect cell lines Journal Article

  

Authors:	Joosten, C. E.; Cohen, L. S.; Ritter, G.; Batt, C. A.; Shuler, M. L.
Article Title:	Glycosylation Profiles of the human colorectal cancer A33 antigen naturally expressed in the human colorectal cancer cell line SW1222 and expressed as recombinant protein in different insect cell lines
Abstract:	The A33 antigen is a cell surface glycoprotein expressed in human gastrointestinal epithelium and in 95% of colorectal cancers. We have compared the N-linked glycosylation profile of A33 antigen naturally expressed in a human colorectal cancer cell line with recombinant human A33 antigen (rA33) produced in insect cell culture using the baculovirus expression vector. N-Linked glycans were enzymatically released from the protein, and glycan composition was analyzed by HPLC. In three insect cell lines tested (Sf-21, Tn5B1-4, and Tn-4s), glycosylation of rA33 was dominated by high mannose structures (M 5Gn2 to M9Gn2; 78-95% of total N-linked glycans), with M8-Gn2 being the single most abundant glycoform. A33 antigen naturally expressed in the SW1222 human colon cancer cell line (A33) also possessed a high abundance of high mannose glycans (72%). No complex glycosylation was detected on rA33 expressed in insect cells. Natural A33 was galactosylated to a small extent (6%). These results illustrate a case of similar glycosylation of a glycoprotein between a recombinant version produced in insect cell culture and its counterpart naturally expressed in human cell culture.
Keywords:	proteins; animal; cytology; metabolism; animals; pathology; cell line, tumor; colorectal neoplasms; blotting, western; immunology; antigens; cell culture; tumors; membrane glycoproteins; colorectal tumor; recombinant proteins; membrane protein; recombinant protein; tumor cell line; western blotting; glycosylation; high performance liquid chromatography; chromatography, high pressure liquid; enzymes; polyacrylamide gel electrophoresis; electrophoresis, polyacrylamide gel; insect; insects; vectors; unidentified baculovirus; insecta; humans; human; article; gpa33 protein, human; gastrointestinal epithelium; glycosylation profiles
Journal Title:	Biotechnology Progress
Volume:	20
Issue:	4
ISSN:	8756-7938
Publisher:	American Chemical Society
Date Published:	2004-07-01
Start Page:	1273
End Page:	1279
Language:	English
DOI:	10.1021/bp034378n
PROVIDER:	scopus
PUBMED:	15296462
DOI/URL:	http://www.scopus.com/inward/record.url?eid=2-s2.0-4043176166&partnerID=40&md5=d27adaaf81e8f702dec4fd87fb926b87
Notes:	Biotechnol. Prog. -- Cited By (since 1996):7 -- Export Date: 16 June 2014 -- CODEN: BIPRE -- Source: Scopus

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