Structural differences among monoclonal antibodies with distinct fine specificities and kinetic properties

T. B. Lavoie, S. Mohan, C. A. Lipschultz, Jean-Charles B. Grivel, Y. Li, C. R. Mainhart, L. N W Kam-Morgan, W. N. Drohan, S. J. Smith-Gill

Research output: Contribution to journalArticle

29 Citations (Scopus)

Abstract

The mAbs HyHEL-8, HyHEL-26 (HH8, and HH26, respectively) recognize epitopes on hen egg-white lysozyme (HEL) highly overlapping with the structurally defined HH10 epitope, while the structurally related XRPC-25 is specific for DNP and does not bind HEL. All four Abs appear to use the same V(k)23 germ line gene, and all but HH8 use the same V(H)36-60 germ line gene. Of the three anti-HEL Abs, the sequences of HH26 variable regions are closest to those encoded by the respective germ line sequences. HH8 utilizes a different member of the V(H)36-60 gene family. Thus, the same V(k) and V(H) genes, combined with somatically derived sequence differences, are used to recognize the unrelated Ags HEL and DNP. In contrast, different V(H)36- 60 germ line genes are used to bind the same antigen (e.g. HH8 vs HH10 and HH26). While the affinities of HH10, HH8, and HH26 for HEL vary by less than 10-fold, their affinities for mutated Ag vary over several orders of magnitude. Analyses of Fab binding kinetics with natural species variants and site-directed mutants of lysozyme indicate that these cross-reactivity differences reflect the relative sensitivities of both the association and dissociation rates to antigenic mutation: HH8 has relatively mutation-insensitive association and dissociation rates, HH10 has a relatively mutation-sensitive association rate but more variable dissociation rates, and HH26 has variable association and dissociation rates. Only a few amino acid differences among the antibodies produce the observed differences in the robustness of the association and dissociation rates. Our results suggest that association and dissociation rates and mutation sensitivity of these rates may be independently modulated during antibody repertoire development. (C) Published by Elsevier Science Ltd.

Original languageEnglish
Pages (from-to)1189-1205
Number of pages17
JournalMolecular Immunology
Volume36
Issue number17
DOIs
Publication statusPublished - Dec 1999
Externally publishedYes

Fingerprint

Egg White
Germ Cells
Monoclonal Antibodies
Genes
Mutation
Epitopes
Antibodies
Mutation Rate
Muramidase
hen egg lysozyme
Antigens
Amino Acids

Keywords

  • Affinity maturation
  • Antibodies
  • Antigen binding
  • Antigen-antibody interactions
  • Binding kinetics

ASJC Scopus subject areas

  • Molecular Biology
  • Immunology

Cite this

Structural differences among monoclonal antibodies with distinct fine specificities and kinetic properties. / Lavoie, T. B.; Mohan, S.; Lipschultz, C. A.; Grivel, Jean-Charles B.; Li, Y.; Mainhart, C. R.; Kam-Morgan, L. N W; Drohan, W. N.; Smith-Gill, S. J.

In: Molecular Immunology, Vol. 36, No. 17, 12.1999, p. 1189-1205.

Research output: Contribution to journalArticle

Lavoie, TB, Mohan, S, Lipschultz, CA, Grivel, J-CB, Li, Y, Mainhart, CR, Kam-Morgan, LNW, Drohan, WN & Smith-Gill, SJ 1999, 'Structural differences among monoclonal antibodies with distinct fine specificities and kinetic properties', Molecular Immunology, vol. 36, no. 17, pp. 1189-1205. https://doi.org/10.1016/S0161-5890(99)00130-3
Lavoie, T. B. ; Mohan, S. ; Lipschultz, C. A. ; Grivel, Jean-Charles B. ; Li, Y. ; Mainhart, C. R. ; Kam-Morgan, L. N W ; Drohan, W. N. ; Smith-Gill, S. J. / Structural differences among monoclonal antibodies with distinct fine specificities and kinetic properties. In: Molecular Immunology. 1999 ; Vol. 36, No. 17. pp. 1189-1205.
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abstract = "The mAbs HyHEL-8, HyHEL-26 (HH8, and HH26, respectively) recognize epitopes on hen egg-white lysozyme (HEL) highly overlapping with the structurally defined HH10 epitope, while the structurally related XRPC-25 is specific for DNP and does not bind HEL. All four Abs appear to use the same V(k)23 germ line gene, and all but HH8 use the same V(H)36-60 germ line gene. Of the three anti-HEL Abs, the sequences of HH26 variable regions are closest to those encoded by the respective germ line sequences. HH8 utilizes a different member of the V(H)36-60 gene family. Thus, the same V(k) and V(H) genes, combined with somatically derived sequence differences, are used to recognize the unrelated Ags HEL and DNP. In contrast, different V(H)36- 60 germ line genes are used to bind the same antigen (e.g. HH8 vs HH10 and HH26). While the affinities of HH10, HH8, and HH26 for HEL vary by less than 10-fold, their affinities for mutated Ag vary over several orders of magnitude. Analyses of Fab binding kinetics with natural species variants and site-directed mutants of lysozyme indicate that these cross-reactivity differences reflect the relative sensitivities of both the association and dissociation rates to antigenic mutation: HH8 has relatively mutation-insensitive association and dissociation rates, HH10 has a relatively mutation-sensitive association rate but more variable dissociation rates, and HH26 has variable association and dissociation rates. Only a few amino acid differences among the antibodies produce the observed differences in the robustness of the association and dissociation rates. Our results suggest that association and dissociation rates and mutation sensitivity of these rates may be independently modulated during antibody repertoire development. (C) Published by Elsevier Science Ltd.",
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AU - Lavoie, T. B.

AU - Mohan, S.

AU - Lipschultz, C. A.

AU - Grivel, Jean-Charles B.

AU - Li, Y.

AU - Mainhart, C. R.

AU - Kam-Morgan, L. N W

AU - Drohan, W. N.

AU - Smith-Gill, S. J.

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AB - The mAbs HyHEL-8, HyHEL-26 (HH8, and HH26, respectively) recognize epitopes on hen egg-white lysozyme (HEL) highly overlapping with the structurally defined HH10 epitope, while the structurally related XRPC-25 is specific for DNP and does not bind HEL. All four Abs appear to use the same V(k)23 germ line gene, and all but HH8 use the same V(H)36-60 germ line gene. Of the three anti-HEL Abs, the sequences of HH26 variable regions are closest to those encoded by the respective germ line sequences. HH8 utilizes a different member of the V(H)36-60 gene family. Thus, the same V(k) and V(H) genes, combined with somatically derived sequence differences, are used to recognize the unrelated Ags HEL and DNP. In contrast, different V(H)36- 60 germ line genes are used to bind the same antigen (e.g. HH8 vs HH10 and HH26). While the affinities of HH10, HH8, and HH26 for HEL vary by less than 10-fold, their affinities for mutated Ag vary over several orders of magnitude. Analyses of Fab binding kinetics with natural species variants and site-directed mutants of lysozyme indicate that these cross-reactivity differences reflect the relative sensitivities of both the association and dissociation rates to antigenic mutation: HH8 has relatively mutation-insensitive association and dissociation rates, HH10 has a relatively mutation-sensitive association rate but more variable dissociation rates, and HH26 has variable association and dissociation rates. Only a few amino acid differences among the antibodies produce the observed differences in the robustness of the association and dissociation rates. Our results suggest that association and dissociation rates and mutation sensitivity of these rates may be independently modulated during antibody repertoire development. (C) Published by Elsevier Science Ltd.

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