title: Preparation and Characterization of a Monoclonal Antibody Against Morphine.
abstract: A monoclonal antibody (MAb) was produced by immunization of a BALB/c mouse with a conjugated morphine C6-hemisuccinated derivative (MHS) to cationized bovine serum albumin (cBSA). The hybridoma clones were screened by indirect ELISA using MHS-BSA. The best hybridoma clone was subcloned thrice by limiting dilution. This hybridoma was found to be of IgG2b class and subclass and contained lambda light chain. The affinity of the MAb to morphine was obtained 2.8×10(9) M(-1). The titer of the cell culture supernatant was at least 1:800. The MAb was cross-reacted with codeine (100%) and apomorphine (16.5%), but not with heroin, naloxone, naltrexone, or papaverine. Morphine was conjugated to HRP using a mixed anhydride method and a direct competitive ELISA was designed using anti-morphine MAb. The assay was sensitive over the 50 ng/mL to 5 μg/mL concentration range. In conclusion, this MAb is useful for the development of immunoassays to measure morphine in urine.
authors: 1 Department of Immunology, School of Medicine, International Campus, Shahid Sadoughi University of Medical Sciences , Yazd, Iran .
authors: Susan
authors: S
authors: Kashanian
authors: 1 Department of Immunology, School of Medicine, International Campus, Shahid Sadoughi University of Medical Sciences , Yazd, Iran .
authors: Ali
authors: A
authors: Shams
authors: 2 Department of Biochemistry, School of Medicine, Tehran University of Medical Sciences , Tehran, Iran .
authors: Hossein
authors: H
authors: Ghahremani
authors: 2 Department of Biochemistry, School of Medicine, Tehran University of Medical Sciences , Tehran, Iran .
authors: Maliheh
authors: M
authors: Paknejad
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000915:Antibody Affinity / Q000276:immunology; D001058:Apomorphine / Q000276:immunology; D003061:Codeine / Q000276:immunology; D003429:Cross Reactions / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D006825:Hybridomas / Q000276:immunology; D007114:Immunization / Q000379:methods; D007118:Immunoassay / Q000379:methods; D007074:Immunoglobulin G / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D009020:Morphine / Q000276:immunology*
chemical_list: D000911:Antibodies, Monoclonal; D007074:Immunoglobulin G; D009020:Morphine; D001058:Apomorphine; D003061:Codeine