David S. Wrenn

[13] Immunology of elastin

Publisher Summary This chapter describes different methods for the preparation of polyclonal and monoclonal antibodies to insoluble or solubilized elastin and will detail their use in a variety of immunological procedures. Antisera have been prepared to insoluble and solubilized elastin using a variety of antigen preparations and animal hosts. The quantities of antigen used to produce these antisera are widely variable and there are insufficient data available for comparison of antibody response at each dose. Accordingly, there is no optimum concentration of elastin that should be used to immunize the host animal. It is important to note that antibodies to insoluble or solubilized forms of elastin may not form precipitable complexes. Therefore, immunoelectrophoresis or immunodiffusion is not the most effective way to monitor antiserum titer or to quantify the amounts of antigen in experimental samples. Passive hemagglutination and competitive protein binding assays are more useful for characterization of antisera and antigen quantification.

Characterization of elastin protein and mRNA from salmonid fish (Oncorhynchus kisutch)

1. Elastin was isolated from the bulbus arteriosus of a salmonid fish. Monoclonal and polyclonal antibodies, elicited against a CNBr digest of this protein, immunoprecipitated a polypeptide of Mr 43,000 from fish cell culture medium. 2. Cell-free translation of salmon poly A+ RNA produced a protein of approximately 43 kD that was immunoprecipitated with anti-elastin antibodies. The corresponding mRNA had an approximate Mr of 2 kb. 3. Despite similarities in amino acid composition, the differences in Mr between mammalian and salmon mRNA and protein suggest a divergence of fish and higher vertebrate elastins from an earlier ancestral gene.