Monoclonal antibodies to human thyroglobulin: production and characterization

Human thyroglobulin was used as an antigen for the development of monoclonal antibodies by the hybridoma technique. Spleen cells of BALB/c mice immunized with human thyroglobulin were fused with SP2/0 mouse myeloma cells. Seven clones secreting specific monoclonal antibodies to thyroglobulin were established. Two of these monoclonal antibodies have been purified and characterized. Their equilibrium association constants (Ka) as determined by Scatchard analysis were 0.24 X 10(11) L/M and 1.4 X 10(11) L/M respectively. The specificity of both these antibodies was validated by immunohistochemical staining of human tissues (normal human thyroid, brain, salivary gland, skeletal and smooth muscle, mucous membrane, parathyroid, adrenal) obtained from autopsy material. Only follicles and follicular cells of thyroid tissue were stained by both the monoclonal antibodies. H10 I monoclonal antibody was used for constructing a standard curve for in vitro immunoassay using a solid phase ELISA technique. The minimum amount detectable was 7.8 ng/ml. Thirty six sera from patients of various thyroid disorders were evaluated using ELISA and compared with conventional RIA. A good agreement was seen (r = 0.92) between the two techniques. These specific monoclonal antibodies may prove to be valuable for in vitro immunoassays and in vivo immunoscintigraphy.