Journal
JOURNAL OF BIOMEDICINE AND BIOTECHNOLOGY
Volume -, Issue -, Pages -Publisher
HINDAWI PUBLISHING CORPORATION
DOI: 10.1155/2012/597647
Keywords
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Funding
- Higher Education Research Promotion and National Research University Project of Thailand
- Office of the Higher Education Commission, through the Center of Excellence in Specific Health Problems in Greater Mekong Subregion Cluster (SHeP-GMS) [NRU 542022]
- Centre for Research and Development of Medical Diagnostic Laboratories (CMDL), Khon Kaen University (KKU)
- KKU through SHeP-GMS
- [H-2533-Ph.d-04]
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Major histocompatibility complex class I chain-related gene A (MICA) is an NKG2D ligand that is over-expressed under cellular stress including cancer transformation and viral infection. High expression of MICA in cancer tissues or patients' sera is useful for prognostic or follow-up markers in cancer patients. In this study, phage display technology was employed to improve antigen-binding activities of anti-MICA monoclonal antibodies (WW2G8, WW6B7, and WW9B8). The 12 amino acid residues in the complementarity determining regions (CDRs) on the V domain of the heavy chain CDR3 (HCDR3) of these anti-MICA antibodies were modified by PCR-random mutagenesis, and phages displaying mutated anti-MICA Fab were constructed. After seven rounds of panning, five clones of phages displaying mutant anti-MICA Fab which exhibited 3-7-folds higher antigen-binding activities were isolated. Two clones of the mutants (phage-displayed mutant Fab WW9B8.1 and phage-displayed mutant Fab WW9B8.21) were confirmed to have antigen-binding specificity for cell surface MICA proteins by flow cytometry. These phage clones are able to recognize MICA in a native formaccording to positive results obtained by indirect ELISA and flow cytometry. Thus, these phage particles could be potentially used for further development of nanomedicine specifically targeting cancer cells expressing MICA proteins.
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