Improvement of the performance of an immunoaffinity extraction method via region-specific immobilization of IgG

Lin Li1, Jin Yan, Mei-Ping Zhao

  • 1Institute of Analytical Chemistry, College of Chemistry and Molecular Engineering, Peking University, Beijing 100871, China.

Using papaverine as a model target, an immunoaffinity column of high selectivity and binding capacity was prepared by utilizing covalent linkage between the Fc portion of IgG and the surface of Sepharose 4B support. Compared with the commonly used random coupling method, the binding capacity of the region-specific immobilized antibodies was increased from 0.04 to 0.2 mol of antigens/mol of antibodies and a much larger concentration factor was thus achieved. The obtained immunoaffinity column has been successfully used in pretreatment of pericarpium papaveris samples. The method offers an improved approach to immunoaffinity extraction that should be useful for purification and concentration of other targeted compounds in highly complex mixture.

Related Concept Videos

Immunoprecipitation01:20

Immunoprecipitation

Immunoprecipitation, or IP, is a widely used technique that employs protein-antibody interactions to isolate proteins or protein complexes in their native state for studying protein-protein interactions, quaternary structures, or supramolecular complexes. Various modifications of the technique, including chromatin IP, cross-linking IP, and fluorescence IP, are commonly used.
Chromatin Immunoprecipitation
Chromatin immunoprecipitation, also known as ChIP, is used to study protein-DNA or...
Immunocytochemistry and Immunohistochemistry01:22

Immunocytochemistry and Immunohistochemistry

Immunocytochemistry (ICC) and immunohistochemistry (IHC) are techniques that use antibodies to check for specific proteins or antigens in a sample. The technique was first published by Albert Coons in 1941 to detect the presence of pneumococcal antigen in tissue sections from mice infected with Pneumococcus. Immunocytochemistry helps localization of proteins or antigens in individual cells like blood cells, stem cells, etc., while immunohistochemistry does the same for tissue samples.
These...
Immunogold Electron Microscopy01:20

Immunogold Electron Microscopy

Immunoelectron microscopy utilizes immunogold labeling of endogenous proteins with specific antibodies to detect and localize these proteins in cells and tissues. The procedure provides insights into the distribution and quantification of protein under different stimulation conditions offering clues about their functions. Conjugating highly electron-dense gold particles with primary or secondary antibodies allow antigen detection on and within cells, with high resolution and specificity.