Production, Characterization, and Functional Analysis of Newly Established CD99 Monoclonal Antibodies MT99/1 and MT99/2

Abstract

The leukocyte surface molecule CD99 is an integral membrane glycoprotein encoded by the E2/MIC2 gene. This molecule is broadly expressed on cells of the hematopoietic system and displays two surface forms, a long 32 kDa form and a short 28 kDa form. While the complete function of the CD99 molecule is unclear, it has been reported to be involved in regulation of cell adhesion, migration, and apoptosis. Thus, several CD99 monoclonal antibodies (MAbs) have been generated for biochemical and functional studies of the CD99 molecule. In the present study two CD99 MAbs, MT99/1 and MT99/2, were produced. The MAbs recognized different epitopes of the CD99 molecule. MAb MT99/1, but not MT99/2, was appropriate for biochemical characterization. Binding of MAb MT99/1 with its epitope led to the induction of cell adhesion and apoptosis. The generated MAbs can be used for future study of the function and mechanism of the CD99 molecule, including its role in the immune system, and may have application in tumor diagnosis and treatment.

Protein expression and gene promoter hypermethylation of CD99 in transitional cell carcinoma of urinary bladder

“…The MIC2 gene product CD99 was a 32-kD cell surface antigen with broad cellular expression, but its clear biological function and signaling pathway are not fully understood (Moreira et al 2008;Khunkaewla et al 2007). It is known that CD99 is originally described as a diagnostically useful marker for Ewing sarcoma and involved in cell-cell adhesion during hematopoietic cell diVerentiation (Gil et al 2002), apoptosis of immature thymocytes (Rochard et al 2004), maintenance of cellular morphology (Cerisano et al 2004), and transport of transmembrane proteins (Vestweber 2007).…”

A modified hybridoma technique for production of monoclonal antibodies having desired isotypes

“…In addition, purification of IgG mAbs is simpler than for the IgM isotype. However, in the conventional hybridoma technique, instead of obtaining the intended IgG mAbs, some mAbs of IgM isotype are always undecided obtained (Chiampanichayakul et al 2006;Khunkaewla et al 2007;our unpublished observations).…”

Generation of a novel Antibody-Drug Conjugate targeting endosialin: potent and durable antitumor response in sarcoma

“…After additional seven days, mice were boosted intravenously with the immunogen, and spleens were removed for cell fusion 3 days later. Somatic cell hybrids were prepared by fusion of immune splenocytes with the murine nonsecreting myeloma cell line NS-1 by standard procedures [ 24 ]. Hybridoma cell lines expressing antibodies recognizing the ECD of human recombinant endosialin in ELISA were subcloned four to six times to ensure that the cell line was monoclonal and stable.…”