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Characterizing High Affinity Antigen/Antibody Complexes by Kinetic and Equilibrium Based Methods

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Book cover Current Trends in Monoclonal Antibody Development and Manufacturing

Part of the book series: Biotechnology: Pharmaceutical Aspects ((PHARMASP,volume XI))

Abstract

The development and application of monoclonal antibodies as drugs requires accurate and precise characterization of the antigen/antibody binding constants. Anticipating the required affinity for a therapeutically efficacious monoclonal antibody (mAb) is complex. Generally, the equilibrium dissociation constant for the antigen/mAb complex should be less than 100–1,000 picomolar (pM), depending upon the nature of the target, the desired function of the antibody, and the localized concentration of the antigen in the diseased tissue, among other factors. Measurement of the equilibrium dissociation constant (K D), association rate constant (k a), and dissociation rate constant (k d) for these high affinity antibodies is difficult because of three independent reasons: (1) the time for the antigen/antibody complex to reach equilibrium can be very long, on the order of days (2) usually, the k d for such a tight complex is extremely low, requiring long periods of data collection in order to discern enough information to predict complex stability, and (3) in cases where the k d is easily measurable (greater than 5×10−4s−1), the k a can be very fast, greater than 1×107M−1s−1.

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Abbreviations

pM:

picomolar

nM:

nanomolar

fM:

femtomolar

mAb:

monoclonal antibody

KD :

equilibrium dissociation constant

ka :

association rate constant

kd :

dissociation rate constant

Fc:

Flow cell

RU:

resonance units

Da:

Dalton

cy5:

Indodicarbocyanine

pAb:

polyclonal antibody

surfactant P-20:

poly(oxyethylene)(20)-sorbitane monolaureate

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Acknowledgments.

The computer algorithm for the simulation of the time to equilibrium for a 1:1 reversible interaction was graciously provided to us by Steve Lackie and Tom Glass of Sapidyne, Inc, Boise, ID.

Author information

Authors and Affiliations

  1. Amgen Fremont, Inc., Fremont, CA, USA

    Andrew W. Drake

  2. Center for Biomolecular Interaction Analysis, University of Utah, Salt Lake City, UT, USA

    David G. Myszka

  3. Takeda San Francisco, S. San Francisco, CA, USA

    Scott L. Klakamp

Authors
  1. Andrew W. Drake
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  2. David G. Myszka
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  3. Scott L. Klakamp
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Corresponding author

Correspondence to Scott L. Klakamp .

Editor information

Editors and Affiliations

  1. Genentech Inc., DNA Way 1, San Francisco, 94080, U.S.A.

    Steven J. Shire

  2. Omeros Corporation, Fifth Ave Ste 2600 1420, Seattle, 98101, U.S.A.

    Wayne Gombotz

  3. Boehringer Ingelheim Pharma KG, Birkendorfer Str. 65, Biberach, 88397, Germany

    Karoline Bechtold-Peters

  4. Genentech, Inc., DNA Way 1, South San Francisco, 94080, U.S.A.

    James Andya

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© 2010 American Association of Pharmaceutical Scientists

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Drake, A.W., Myszka, D.G., Klakamp, S.L. (2010). Characterizing High Affinity Antigen/Antibody Complexes by Kinetic and Equilibrium Based Methods. In: Shire, S., Gombotz, W., Bechtold-Peters, K., Andya, J. (eds) Current Trends in Monoclonal Antibody Development and Manufacturing. Biotechnology: Pharmaceutical Aspects, vol XI. Springer, New York, NY. https://doi.org/10.1007/978-0-387-76643-0_11

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