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In Vivo Immunofluorescence Localization for Assessment of Therapeutic and Diagnostic Antibody Biodistribution in Cancer Research
Published on: September 16, 2019
[Monoclonal antibodies]
1Department of Obstetrics and Gynecology, Hyogo Medical College.
Since the first successful application of somatic cell fusion by Kohler and Milstein to generate hybridomas secreting monoclonal antibodies (Mabs), this hybridoma technique has been widely applied for production of Mabs against a wide variety of antigens. In the lecture, general procedures for establishment of hybridomas and characteristic properties of Mabs were briefly explained and followed by presentation of our clinical studies using Mabs produced in our laboratory to hCG, sperm and tumor cells. 1. Monoclonal antibodies to hCG. Three hybridomas (5D4, 6E4, 2F8) were established by fusing mouse myeloma cells (P3U1) with spleen cells from BALB/c mice immunized with partially purified hCG. Mabs were obtained either from hybridoma culture media or from ascites of mice inoculated the hybridoma cells intraperitoneally. Three Mabs obtained were all IgG1 and each had a unique binding specificity to hCG, hCG-beta, hCG-alpha and LH. They were applied for development of specific, sensitive and easy to perform hCG assays. A reverse passive hemagglutination assay of urinary hCG with a sensitivity of 12.5 IU/l and a sandwich-enzyme immunoassay of serum hCG with a sensitivity of 0.1 mIU/ml were presented. 2. Monoclonal antibodies to human spermatozoa. For elucidation of mechanisms for induction of auto- or iso-sperm immunization and infertility by antisperm antibodies, antigen analysis of seminal components was essential. For this purpose, several Mabs with strong sperm immobilizing (SI) and agglutinating (SA) activities were generated in mice, rats and humans and the corresponding antigens were analysed.(ABSTRACT TRUNCATED AT 250 WORDS)
Since the first successful application of somatic cell fusion by Kohler and Milstein to generate hybridomas secreting monoclonal antibodies (Mabs), this hybridoma technique has been widely applied for production of Mabs against a wide variety of antigens. In the lecture, general procedures for establishment of hybridomas and characteristic properties of Mabs were briefly explained and followed by presentation of our clinical studies using Mabs produced in our laboratory to hCG, sperm and tumor cells. 1. Monoclonal antibodies to hCG. Three hybridomas (5D4, 6E4, 2F8) were established by fusing mouse myeloma cells (P3U1) with spleen cells from BALB/c mice immunized with partially purified hCG. Mabs were obtained either from hybridoma culture media or from ascites of mice inoculated the hybridoma cells intraperitoneally. Three Mabs obtained were all IgG1 and each had a unique binding specificity to hCG, hCG-beta, hCG-alpha and LH. They were applied for development of specific, sensitive and easy to perform hCG assays. A reverse passive hemagglutination assay of urinary hCG with a sensitivity of 12.5 IU/l and a sandwich-enzyme immunoassay of serum hCG with a sensitivity of 0.1 mIU/ml were presented. 2. Monoclonal antibodies to human spermatozoa. For elucidation of mechanisms for induction of auto- or iso-sperm immunization and infertility by antisperm antibodies, antigen analysis of seminal components was essential. For this purpose, several Mabs with strong sperm immobilizing (SI) and agglutinating (SA) activities were generated in mice, rats and humans and the corresponding antigens were analysed.(ABSTRACT TRUNCATED AT 250 WORDS)
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