Production and Selection of Monoclonal Antibodies for Detection of Small Molecular Weight Compounds.
Summary rept. 1 Oct 86-30 Sep 87,
CALIFORNIA UNIV OAKLAND NAVAL BIOSCIENCES LAB
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Production of useful anti-hapten monoclonal antibodies requires integrating carefully chosen immunization, selection, screening, and cell management techniques. This talk will survey principles and methods that many laboratories use but few systematically evaluate. Design and hapten density of the conjugates used for immunizing and screening are critical for successfully hybridoma production. Immunization protocols should be set up to avoid causing suppression of hapten-specific responses. Unrelated mouse strains should be immunized because various inbred lines express different immuno-globulin variable-region genes, and other immunoregulatory genes affect response to hapten. Adjuvants can be chosen with respect to their behavior as immunostimulators, solvents, and depots for hapten-carrier complexes. Immune responses modifiers may be used to stimulate helper T-cell responses or inhibit suppressor T-cell responses to the hapten, or to promote antibody production in B cells. Use of conditioned media instead of feeder cells simplifies hybridoma culture and allows growth at low cell densities. This maximizes detectability and minimizes loss of good clones.
- Medicine and Medical Research
- Genetic Engineering and Molecular Biology