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Isoelectric focusing analysis of antibody clonotype changes occurring during immune responses using immobilized pH gradients
Author(s) -
Knisley Keith A.,
Rodkey L. Scott
Publication year - 1988
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.1150090406
Subject(s) - isoelectric focusing , antibody , immune system , toxoid , isoelectric point , chromatography , affinity chromatography , chemistry , biology , tetanus , antibody response , antiserum , immunology , microbiology and biotechnology , biochemistry , vaccination , immunization , enzyme
Serum was collected from rabbits at 2‐day intervals following a single injection with tetanus toxoid or at weekly intervals following multiple injections with Micrococcus lysodeikticus cell walls. These sera were analyzed for the presence of individual clonotypes of specific anti‐tetanus or anti‐micrococcal antibodies by isoelectric focusing in immobilized pH gradients with added carrier ampholytes followed by affinity immunoblotting. The affinity immunoblots obtained clearly defined both the rapid disappearance and late appearance of distinct subsets of antibody clonotypes during the response. These data demonstrate the application of affinity immunoblotting combined with immobilized pH gradients for detecting the subtle changes in specific antibody clonotype patterns which occur during an immune response.