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Production of Monoclonal Antibodies to Bovine Leukocyte Cell-Surface Components

Mastitis Research Laboratory, North Louisiana Hill Farm Experiment Station, Louisiana State University, Route 1, Box 10, Homer 71040

ABSTRACT

Fusion of murine myeloma cells with syngeneic spleen lymphocytes has led to development of hybridomas secreting antibodies. Hybrid cells retain the immortality and clonability of myeloma parents as well as the antibody-producing property of lymphocytes. Specificity of monoclonal antibody produced is based on the one lymphocyte-one antibody phenomenon and represents the most effective process for producing specific antisera. Spleen cells from mice immunized with desired antigen are hybridized with nonsecreting mouse myeloma cells. Resulting hybrids are cloned and culture fluids tested for specific antibody activity to the antigen. Positive clones are cultivated in vitro and injected into mice for monoclonal antibody production. This technology has been extended to the bovine species to obtain monoclonal antisera to immunoglobulins and cell-surface components of leukocytes for study of mammary gland immunity. Recent progress in monoclonal research has led to interspecific fusion of murine myelomas with bovine lymphocytes, resulting in hybridomas that produce monoclonal bovine immunoglobulins. Monoclonal antibodies will be useful in investigations applicable to bovine research including purification of immunoglobulins, determining immunoglobulin concentration in colostrum and milk, reference reagents for bovine serology, antibody localization in tissue, gene sequencing, characterizing histo-compatibility antigens, distinguishing and quantitating cell types in blood, milk, and udder tissue, and elucidating role of cell subpopulations in the immune response.

¹ Milk Secretion and Mastitis Laboratory, USDA, ASI-SEA-AR, BARC-East, Beltsville, MD 20705.

² Milk Secretion and Mastitis Laboratory, USDA, ASI-SEA-AR, BARC-East, Beltsville, MD 20705.

³ Biology Department, Amherst College, Amherst, MA 01002.

⁴ Biology Department, Amherst College, Amherst, MA 01002.