Production and Characterization of Anti-Her2 Monoclonal Antibodies

Document Type : Original Article

Authors

1 Science and Research Branch, Islamic Azad University

2 Reproductive Biotechnology Research Center, Avicenna Research Institute

3 Immunology Research Center, Faculty of Medicine, Iran University of Medical Sciences

4 Cellular and Molecular Biology Department, Faculty of Biology, Paradise of Science, Tehran University

5 Institute Research Avicenna, Center Research N

6 Monoclonal Antibody Research Center, Avicenna Research Institute

7 Biochemistry Department, Faculty of Medical Sciences, Tarbiat Modarres University

Abstract

Objective: Breast cancer is the most common cancer among women in the world.
Early diagnosis of this cancer is a key element for its treatment. One of the approaches
for diagnosis of breast cancer is detection of its tumour-associated markers. Hence,
Her2 has been the main focus of the researches in the field.
Materials and Methods: For diagnosis of Her2 overexpression, monoclonal
antibodies (mAb) reacting against Her2 were produced in this study. For this
purpose, two peptides from extracellular domain of Her2 were selected and the
mAbs reacting against them were produced by hybrodoma technology. Reactivity
of these antibodies were then evaluated in different immunological assays including
ELISA, Immunoflurescence (IF), western blot (WB) and immunoprecipitation (IP).
Results: Total of 5 clones were produced from two separate fusions, and antibody
isotyping revealed that all clones were IgM. These mAbs showed appropriate
reactivities in the following assays: ELISA, immunofluresence by staining of breast
cancer cell line (SKBR3), WB and IP by detecting the 185 KD band of Her2.
Conclusion: In conclusion, it seems that the mAbs are useful diagnostic tools for
detection of Her2 expression in patients with breast cancer.

Keywords