%0 Journal Article %1 citeulike:763096 %A Nygren, H. %A Czerkinsky, C. %A Stenberg, M. %D 1985 %J J Immunol Methods %K dissociation affinity analysis monovalent %N 1 %P 87--95 %T Dissociation of antibodies bound to surface-immobilized antigen. %U http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=3908564 %V 85 %X The dissociation of antibodies bound to surface-immobilized antigen was investigated by the ELISA, using a hapten (TNP) as antigen. Antibody binding was found to be stable, and no half-time of dissociation could be defined within 69 h. The role of the bivalence of antibodies and the difference between a homogeneous and a heterogeneous reaction was investigated by comparing the dissociation rate of antigen-antibody complexes formed by monovalent Fab fragments from surface-immobilized antigen and the dissociation rate of TNP-lysine from antibodies in a homogeneous liquid phase. Fab fragments were found to dissociate with a half-time value of about 16 h, whereas the homogeneous binding of TNP-antibody dissociated with a half-time of less than 4 h, indicating that both the bivalence of antibodies and the solid phase contributed to the stability of surface-bound antigen-antibody complexes. Qualitative differences between antibodies produced by different clones in a polyclonal antibody response to TNP was investigated by a spot assay. The results indicated that a minority of the antibodies produced had the capacity of binding practically irreversibly to solid-phase-immobilized antigen. The impact of the results on the interpretation of data from solid-phase assays is discussed together with the biological importance of the findings.

@article{citeulike:763096, abstract = {The dissociation of antibodies bound to surface-immobilized antigen was investigated by the ELISA, using a hapten (TNP) as antigen. Antibody binding was found to be stable, and no half-time of dissociation could be defined within 69 h. The role of the bivalence of antibodies and the difference between a homogeneous and a heterogeneous reaction was investigated by comparing the dissociation rate of antigen-antibody complexes formed by monovalent Fab fragments from surface-immobilized antigen and the dissociation rate of TNP-lysine from antibodies in a homogeneous liquid phase. Fab fragments were found to dissociate with a half-time value of about 16 h, whereas the homogeneous binding of TNP-antibody dissociated with a half-time of less than 4 h, indicating that both the bivalence of antibodies and the solid phase contributed to the stability of surface-bound antigen-antibody complexes. Qualitative differences between antibodies produced by different clones in a polyclonal antibody response to TNP was investigated by a spot assay. The results indicated that a minority of the antibodies produced had the capacity of binding practically irreversibly to solid-phase-immobilized antigen. The impact of the results on the interpretation of data from solid-phase assays is discussed together with the biological importance of the findings.}, added-at = {2006-07-31T20:01:51.000+0200}, author = {Nygren, H. and Czerkinsky, C. and Stenberg, M.}, biburl = {https://www.bibsonomy.org/bibtex/2abf9f4076996b4adc5d20faf148f9597/biblio24}, citeulike-article-id = {763096}, interhash = {9311c098eea283ac288ba3ee20d0a0cd}, intrahash = {abf9f4076996b4adc5d20faf148f9597}, issn = {0022-1759}, journal = {J Immunol Methods}, keywords = {dissociation affinity analysis monovalent}, month = {December}, number = 1, pages = {87--95}, priority = {2}, timestamp = {2006-07-31T20:01:51.000+0200}, title = {Dissociation of antibodies bound to surface-immobilized antigen.}, url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve\&db=pubmed\&dopt=Abstract\&list_uids=3908564}, volume = 85, year = 1985 }