%0 Journal Article %1 citeulike:4545205 %A Allen, D. %A Simon, T. %A Sablitzky, F. %A Rajewsky, K. %A Cumano, A. %D 1988 %J The EMBO journal %K antibody, germinalcenter, immunology %N 7 %P 1995--2001 %T Antibody engineering for the analysis of affinity maturation of an anti-hapten response. %U http://view.ncbi.nlm.nih.gov/pubmed/3138111 %V 7 %X The influence of structural variation, previously observed in a panel of V186.2 VH/V lambda 1-expressing anti-NP antibodies from the secondary response, on the affinity of these antibodies was examined by site-specific mutagenesis and recombinant antibody construction. A tryptophan----leucine exchange at position 33 in the VH segment of all but one of the high-affinity antibodies is the most frequently observed somatic mutation and by itself leads to a 10-fold higher affinity; all other somatic exchanges are irrelevant for affinity selection. In the single case of a high-affinity antibody without this common exchange, high affinity is mediated by a combination of mutations (including a one-codon deletion) in VH and the particular D-JH rearrangement carried by this antibody. The data indicate that the pattern of somatic diversification through hypermutation is shaped by affinity selection, but that only a single point mutation is available in the VH and the VL gene of lambda 1 chain-bearing anti-NP antibodies which by itself leads to an increase of hapten-binding affinity. Based on the analysis of two secondary response antibodies from which somatic mutations in VH and VL have been eliminated, it is also concluded that the recruitment of B cell clones into the pathway of hypermutation involves a mechanism which is not based upon affinity differences towards the antigen.

@article{citeulike:4545205, abstract = {The influence of structural variation, previously observed in a panel of V186.2 VH/V lambda 1-expressing anti-NP antibodies from the secondary response, on the affinity of these antibodies was examined by site-specific mutagenesis and recombinant antibody construction. A tryptophan----leucine exchange at position 33 in the VH segment of all but one of the high-affinity antibodies is the most frequently observed somatic mutation and by itself leads to a 10-fold higher affinity; all other somatic exchanges are irrelevant for affinity selection. In the single case of a high-affinity antibody without this common exchange, high affinity is mediated by a combination of mutations (including a one-codon deletion) in VH and the particular D-JH rearrangement carried by this antibody. The data indicate that the pattern of somatic diversification through hypermutation is shaped by affinity selection, but that only a single point mutation is available in the VH and the VL gene of lambda 1 chain-bearing anti-NP antibodies which by itself leads to an increase of hapten-binding affinity. Based on the analysis of two secondary response antibodies from which somatic mutations in VH and VL have been eliminated, it is also concluded that the recruitment of B cell clones into the pathway of hypermutation involves a mechanism which is not based upon affinity differences towards the antigen.}, added-at = {2009-05-19T18:00:18.000+0200}, author = {Allen, D. and Simon, T. and Sablitzky, F. and Rajewsky, K. and Cumano, A.}, biburl = {https://www.bibsonomy.org/bibtex/24459ebb6057c8e4a6613fceca27b4043/earthfare}, citeulike-article-id = {4545205}, description = {CiteULike: Everyone's library}, interhash = {2f2b23ff0c13b61e214de6e58b061393}, intrahash = {4459ebb6057c8e4a6613fceca27b4043}, issn = {0261-4189}, journal = {The EMBO journal}, keywords = {antibody, germinalcenter, immunology}, month = {July}, number = 7, pages = {1995--2001}, posted-at = {2009-05-19 15:50:39}, priority = {2}, timestamp = {2009-05-19T18:03:27.000+0200}, title = {Antibody engineering for the analysis of affinity maturation of an anti-hapten response.}, url = {http://view.ncbi.nlm.nih.gov/pubmed/3138111}, volume = 7, year = 1988 }