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The Journal of Immunology, 2007, 179, 3841-3850
Copyright © 2007 by The American Association of Immunologists, Inc.
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Kinetic, Affinity, and Diversity Limits of Human Polyclonal Antibody Responses against Tetanus Toxoid

Tine Rugh Poulsen, Per-Johan Meijer, Allan Jensen, Lars S. Nielsen and Peter S. Andersen1

Department for Antibody Discovery, Symphogen A/S, Lyngby, Denmark

Due to technical limitations, little knowledge exists on the composition of Ag-specific polyclonal Ab responses. Hence, we here present a molecular analysis of two representative human Ab repertoires isolated by using a novel single-cell cloning approach. The observed genetic diversity among tetanus toxoid-specific plasma cells indicate that human polyclonal repertoires are limited to the order of 100 B cell clones and hypermutated variants thereof. Affinity and kinetic binding constants are log-normally distributed, and median values are close to the proposed affinity ceilings for positive selection. Abs varied a million-fold in affinity but were restricted in their off-rates with an upper limit of 2 x 10–3 s–1. Identification of Abs of high affinity without hypermutations in combination with a modest effect of hypermutations on observed affinity increases indicate that Abs selected from the naive repertoire are not only of low affinity but cover a relatively large span in affinity, reaching into the subnanomolar range.

The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked advertisement in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

1 Address correspondence and reprint requests to Dr. Peter S. Andersen, Symphogen A/S, Elektrovej Bygning 375, Lyngby, Denmark. E-mail address: psa{at}symphogen.com

2 Abbreviations used in this paper: VH, H chain variable domain; JH, H chain joining region; SPR, surface plasmon resonance; TT, tetanus toxoid; V{kappa}, {kappa}-chain variable domain; J{kappa}, {kappa}-chain joining region.






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