title: Monoclonal antibodies to mouse butyrylcholinesterase.
abstract: Our immunization strategy introduced recombinant mouse butyrylcholinesterase (BChE) to naïve BChE knockout mice. An extraordinarily strong immune reaction gave rise to a whole spectrum of antibodies with different properties. Two selective and highly efficient monoclonal anti-mouse BChE antibodies 4H1 (IgG1) and 4 C9 (IgG2a), with Kd values in the nanomolar range were generated. ELISA detected BChE in as little as 20-50 nl of mouse plasma using 2 μg (4H1) or 4 μg (4C9). Both antibodies cross-reacted with BChE in dog plasma but only 4 H1 reacted with rat BChE, suggesting that the antibodies are targeted towards different epitopes. Surprisingly, neither recognized human BChE. The anti-mouse BChE antibodies were used in immunohistochemistry analysis of mouse muscle where they specifically stained the neuromuscular junction. The antibodies enable visualization of the BChE protein in the mouse tissue, thus complementing activity assays. They can be used to study a long-lasting question about the existence of mixed acetylcholinesterase/BChE oligomers in mouse tissues. Moreover, monoclonal anti-mouse BChE antibodies can provide a simple, fast and efficient way to purify mouse BChE from small amounts of starting material by using a single-step immunomagnetic bead-based protocol.
authors.affiliation: Faculty of Pharmacy, Dept of Pharmacology and Toxicology, Comenius University, Bratislava, Slovakia.
authors.forename: Katarina
authors.initials: K
authors.lastname: Mrvova
authors.forename: Lucia
authors.initials: L
authors.lastname: Obzerova
authors.forename: Emmanuelle
authors.initials: E
authors.lastname: Girard
authors.forename: Eric
authors.initials: E
authors.lastname: Krejci
authors.forename: Anna
authors.initials: A
authors.lastname: Hrabovska
mesh_terms: D000818:Animals; D058846:Antibodies, Monoclonal, Murine-Derived / Q000096:biosynthesis* / Q000378:metabolism; D000915:Antibody Affinity; D000918:Antibody Specificity; D002091:Butyrylcholinesterase / Q000172:deficiency / Q000235:genetics / Q000276:immunology*; D003429:Cross Reactions; D004285:Dogs; D006801:Humans; D007114:Immunization; D051379:Mice; D018345:Mice, Knockout; D051381:Rats; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology; D013045:Species Specificity
chemical_list: D058846:Antibodies, Monoclonal, Murine-Derived; D011994:Recombinant Proteins; D002091:Butyrylcholinesterase