Production of Monoclonal Antibody to Oxytocin for Immunohistochemical Use

Abstract

This study was performed to develop a hybrid cell line secreting antibody to oxytocin for immunohistochemical localization of oxytocin neuron in the brain. Synthetic oxytocin was conjugated to bovine thyroglobulin and used in immunization of Balb/c mice. The spleen cells from immunized mice were fused with NS1 murine myeloma cells using polyethylene glycol, and the hybrid cells were selected by HAT medium. A radiobinding test was used to detect the presence of antibody in the culture fluid of hybrids, and specificity of the antibodies was characterized by both modified enzyme linked immunosorbent assay and immunocytochemical method. Two hybrid cell lines were found to secrete specific anti-oxytocin antibodies. These monospecific antibodies did not cross-react with arginine vasopressin or vasotocin. By light immunocytochemistry these antibodies were able to localize differentially the oxytocin neurons from the vasopressin neurons in the paraventricular nucleus, supraoptic nucleus, median eminence and posterior pituitary gland.