Immunochemistry

The ability to measure the interactions between antigens and antibodies is fundamental to work done by the laboratory. Immunochemical technologies in routine use include

  1. double (Ouchterlony) immunodiffusion
  2. quantitative precipitin reactions
  3. agglutination including passive hemagglutination and latex agglutination
  4. enzyme linked immunosorbent assay (ELISA)
  5. various forms of immunoblot (one dimensional and two dimensional)

A plate reader and plate washer are available for ELISA. Various power packs and electrophoretic transfer equipment are used for immunoblots. Visualization of immunoblots is done with a Bio-Rad ChemiDoc XRS+ imaging system.

Purification and characterization of antigens and antibodies is done with a full array of systems for liquid chromatography and electrophoresis. Antibody purification uses four Isco low pressure liquid chromatography systems. Antigen purification is done with a Bio-Rad high pressure liquid chromatography system.

Antibody labeling is done in house on a routine basis. Labels include various enzymes, fluorophores and biotin/streptavidin. Antibody fragmentation is also done in house to prepare Fab and F(ab')2 fragments.

Highly immunogenic constructs of peptides and polysaccharides are prepared by conjugation of haptens to protein carriers. A variety of coupling chemistries are used to optimize conjugate formation and conjugate immunogenicity.

Imaging of the interactions between antibodies and microbes is a core technology for the laboratory. Several images have been used for cover art for scientific journals and textbooks. Fluorescence microscopy is done with a Zeiss LSM 700 laser scanning microscope.