Development of high-affinity monoclonal antibody using CD44 overexpressed cells as a candidate for targeted immunotherapy and diagnosis of acute myeloid leukemia
Affiliations: [a] National Cell Bank of Iran, Pasteur Institute of Iran, Tehran, Iran | [b] Immunology Department, Hybridoma Laboratory, Pasteur Institute of Iran, Tehran, Iran | [c] Department of Mycobacteriology, Pasteur Institute of Iran, Tehran, Iran
Correspondence:
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Corresponding author: Mohammad Ali Shokrgozar, National Cell Bank of Iran, Pasteur Institute of Iran, Tehran, Iran. Tel.: +98 21 66492595; Fax: +98 21 66492595; E-mail:[email protected];[email protected]
Abstract: AIM: CD44s antigens have been suggested as an efficient biomarker for cancer stem
cells. Current study aimed to develop a hybridoma that producing a high
affinity murine anti-human CD44 monoclonal antibody for early diagnostic
laboratory tests of some cancer. MATERIALS AND METHODS: To make hybridoma against CD44, mice were immunized with MDA-MB-468 cells. Resulted hybridomas using three culture media were screened by indirect
ELISA, then cloned by limiting dilution, and isotype was determined after
obtaining ascitic fluid and antibody purification. RESULTS: We obtained a stable secreting clone, capable of secreting a high-affinity
monoclonal antibody against CD44 protein, IgG2a kappa, with the affinity of
5.4 × 10-8 M without cross-reactivity. CONCLUSION: We could establish a hybridoma in a native form. This stable and
high-affinity anti-CD44 mAb has a potential for diagnostic procedures and
laboratory research. Thus, it could be exploited as a suitable tool for
target-specific diagnosis and even treatment in several cancers.
