|
Hybridoma
Preparation and Characterization of Monoclonal Antibody Against Melatonin
To cite this article:
Mohammad Soukhtanloo, Mohammad Ansari, Maliheh Paknejad, Mohammad Reza Parizadeh, Mohammad Javad Rasaee.
Hybridoma.
June 2008,
27(3): 205-209.
doi:10.1089/hyb.2008.0002.
Mohammad Soukhtanloo Department of Clinical Biochemistry, School of Medicine, Medical Sciences/University of Tehran, Iran. Mohammad Ansari Department of Clinical Biochemistry, School of Medicine, Medical Sciences/University of Tehran, Iran. Maliheh Paknejad Department of Clinical Biochemistry, School of Medicine, Medical Sciences/University of Tehran, Iran. Mohammad Reza Parizadeh Department of Biochemistry and Nutrition, Faculty of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran. Mohammad Javad Rasaee Department of Medical Biotechnology, School of Medical Sciences, Tarbiat Modarres University, Tehran, Iran. Abstract Anti-melatonin monoclonal antibodies (MAb) were prepared following coupling melatonin to bovine serum albumin (BSA) by Mannich reaction. Balb/c mice were immunized via injection of the melatonin-BSA intraperitonally. The spleen cells producing high titer of antibody were fused with myeloma cells of SP2/0 origin. After two limiting dilutions, two stable clones (AS-H10 and AS-D26) exhibiting best properties were selected for further studies. The class and subclass of two MAbs were found to be IgG1 and IgG2a with λ and κ light chains, respectively. Antibodies secreted by these two clones showed high affinity of about 109M−1. Study of the specificity criteria showed that these clones had no cross reactivity with indolic, aromatic, and imidazole ring-containing compounds, and had high specificity towards melatonin. The calibration curve was constructed with a sensitivity range of 10 ng/mL to 10 μg/mL. In conclusion, these MAbs may be useful for immunoassay of melatonin.
|
|
TOC Alert
CiteULike
Delicious
Digg This
Facebook
Newsvine