Development of monoclonal antibodies against bovine mucin core 2 β6 N-acetylglucosaminyltransferase

Cheng Ming Li, Nirmal Joshee, Kenneth B. Adler, Pi Wan Cheng

Research output: Contribution to journalArticle

3 Scopus citations

Abstract

Molecular cloning techniques have been used to produce abundant amounts of recombinant glycosyltransferases for biochemical studies. We recently cloned a cDNA which encoded bovine mucin core 2 β6N-acetylglucosaminyl transferase (C2TF). poly-histidine-C2TF fusion protein was generated from the cloned cDNA in the E. coil Xpress system and used to produce monoclonal antibodies (MAbs). We obtained seven hybridomas which secreted MAbs against bovine C2TF in mouse ascites with titers ranging from 1:1280 to 1:40960 as assessed by immunofluorescence assay (IF). Isotyping revealed that all seven MAbs were IgG (4 IgG1, 2 IgG2b and 1 IgG2a). The affinity constants (M-1) for these MAbs range from 5.4 x 107 to 1.2 x 109. These MAbs recognized bovine C2TF in tissue sections and on Western blottings. Six of these MAbs reacted with human core 2-M enzyme and one with both core 2-L and core 2-M enzymes on Western blottings. Therefore, These antibodies should be useful for further study of bovine and human core 2 enzymes.

Original languageEnglish (US)
Pages (from-to)555-562
Number of pages8
JournalGlycoconjugate Journal
Volume16
Issue number9
DOIs
StatePublished - 1999

Keywords

  • Core 2 N-acetylglucosaminyltransferase
  • Glycosyltransferase
  • Monoclonal antibodies
  • Mucin

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

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