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Characterization of Mono‐ and Polyclonal Antibodies Against Highly Purified Choline Acetyltransferase: A Monoclonal Antibody Shows Reactivity in Human Brain
Author(s) -
OstermannLatif Christel,
Mäder Michael,
Unger Jürgen W.,
Bartke Ilse,
Naujoks Kurt,
Peters Johann H.,
Felgenhauer Klaus
Publication year - 1992
Publication title -
journal of neurochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.75
H-Index - 229
eISSN - 1471-4159
pISSN - 0022-3042
DOI - 10.1111/j.1471-4159.1992.tb09362.x
Subject(s) - monoclonal antibody , polyclonal antibodies , choline acetyltransferase , microbiology and biotechnology , antibody , affinity chromatography , biology , subclass , isoelectric focusing , monoclonal , biochemistry , enzyme , immunology , cholinergic , neuroscience
Choline acetyltransferase (ChAT) from porcine brain was purified by immunoaffinity chromatography, and the highly purified enzyme was subsequently used for immunization of mice and rabbits. After fusion of mouse spleen cells, 32 cultures producing monoclonal antibodies directed against ChAT were detected by an enzyme‐linked immu‐nosorbent assay (ELISA) with immunoaffinity‐purified ChAT. Of these original 32, the most active 11 cultures were cloned and used for ascites production. The 11 clones generated monoclonal antibodies of the immunoglobulin (Ig) M class (three), the IgGl subclass (seven), and the IgG2b subclass (one). The isoelectric points of the antibodies of the IgG class were different in each case. The monoclonal antibodies exhibited different binding characteristics in the above ELISA and on western blots. Two monoclonal antibodies demonstrated excellent immunohistological results with neurons of rat brain and spinal cord. One of them reacted well immu‐nohistochemically with neurons of human brain and also recognized partially purified human placenta ChAT in the ELISA.