Only selected light chains combine with a given heavy chain to confer specificity for a model glycopeptide antigen.
The M and N human blood group glycopeptide Ags are carried on RBCs by glycophorin A. Previous results suggested that the murine humoral immune response against the N, but not the M, Ag is restricted. In addition, these results suggested that particular highly homologous heavy chains might be able to combine promiscuously with various light chains to yield anti-N specificity. To examine this, the current study used Fab phage methodology to couple an array of light chains, obtained from cDNA libraries isolated from immunized mice, to single Fd obtained from N61, N92, and 425/2B hybridomas. Interestingly, for the chimeric Fab to retain M or N specificity, the new light chains needed to belong to the same Vk gene family as the light chain from the parental, hybridoma-derived mAb. In some cases the new light chains modified the Fab affinity and fine specificity. For example, library-derived light chains coupled with the N92 Fd yielded chimeric Fab with increased affinity. In particular, the affinity of these univalent chimeric Fab for the N Ag was equivalent to that of the bivalent parental IgG mAb. Taken together, these results demonstrate that particular structures formed by the light chain V region are required to cooperate with a particular heavy chain V region to create a functional binding site for these glycopeptide Ags. They also demonstrate a lack of heavy chain promiscuity in the formation of murine anti-M and anti-N Abs.
Czerwinski
M
M
Ludwik Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland.
Siemaszko
D
D
Siegel
D L
DL
Spitalnik
S L
SL
D000595:Amino Acid Sequence; D000818:Animals; D000918:Antibody Specificity / Q000235:genetics* / Q000276:immunology; D001483:Base Sequence; D015723:Gene Library; D006020:Glycopeptides / Q000276:immunology*; D006801:Humans; D007143:Immunoglobulin Heavy Chains / Q000235:genetics* / Q000276:immunology; D007147:Immunoglobulin Light Chains / Q000235:genetics* / Q000276:immunology; D008951:MNSs Blood-Group System / Q000276:immunology*; D051379:Mice; D008969:Molecular Sequence Data; D017421:Sequence Analysis
D006020:Glycopeptides; D007143:Immunoglobulin Heavy Chains; D007147:Immunoglobulin Light Chains; D008951:MNSs Blood-Group System