Journal of Mathematical Chemistry

, Volume 52, Issue 2, pp 575–587 | Cite as

Application of the log-normal model for long term high affinity antibody/antigen interactions using Bio-Layer Interferometry

  • Jakob WallnerEmail author
  • Manfred Kühleitner
  • Norbert Brunner
  • Gabriele Lhota
  • Karola Vorauer-Uhl
Original Paper


For more than 50 years, optical biosensors have been used to measure bio-molecular interactions. The most frequently applied binding model to fit biosensor data is the simple 1:1 binding model which requires the stabilization of the association phase to the equilibrium Req and the stabilization of the dissociation phase to the equilibrium zero. However, due to technical limitations many published biosensor measurements are finished before these requirements are fulfilled. In the present study, a long term binding interaction analysis with a monoclonal antibody, namely IgG 2F5 and UG37 a specific antigen with a promising biosensor platform, the Bio-Layer Interferometry, was performed. Data fitting with the simple 1:1 binding model to the association phase was inappropriate and the fitted parameters varied with the concentration and time, which contradicts the theory of the simple 1:1 binding model. Furthermore, extrapolation of the fits with individual times spans compared to 100 % of the obtained data systematically underestimated the actual observed binding curve. Interestingly, an alternative model based on the cumulative distribution function of the log-normal probability distribution remedied the aforementioned problems allowing \(\hbox {K}_\mathrm{L}\) (which is the analog to the affinity constant \(\hbox {K}_\mathrm{D}\)) to be estimated. We further demonstrate that this model fits the biosensor data far better and is essentially less affected by the stabilization of the association phase to the equilibrium (Req) and the stabilization of the dissociation phase to the equilibrium zero. Finally, extrapolation with the log-normal model predicts the actually observed binding curve in a proper manner.


Log-normal model Simple 1:1 interaction model Biosensor Bio-Layer Interferometry Curve fitting Antibody/antigen interactions 



The authors thank Polymun Scientific, Immunbiologische Forschung GmbH (Klosterneuburg, Austria), for kindly providing the recombinant human monoclonal antibody (IgG2F5) and the specific antigen UG37 in pharmaceutical grade.


  1. 1.
    A. Giannetti, B. Koch, M. Browner, J. Med. Chem. 51, 574 (2008)CrossRefGoogle Scholar
  2. 2.
    R.L. Rich, G.A. Papalia, P.J. Flynn, J. Furneisen, J. Quinn, J.S. Klein, P.S. Katsamba, M.B. Waddell, M. Scott, J. Thompson, J. Berlier, S. Corry, M. Baltzinger, G. Zeder-Lutz, A. Schoenemann, A. Clabbers, S. Wieckowski, M.M. Murphy, P. Page, T.E. Ryan, J. Duffner, T. Ganguly, J. Corbin, S. Gautam, G. Anderluh, A. Bavdek, D. Reichmann, S.P. Yadav, E. Hommema, E. Pol, A. Drake, S. Klakamp, T. Chapman, D. Kernaghan, K. Miller, J. Schuman, K. Lindquist, K. Herlihy, M.B. Murphy, R. Bohnsack, B. Andrien, P. Brandani, D. Terwey, R. Millican, R.J. Darling, L. Wang, Q. Carter, J. Dotzlaf, J. Lopez-Sagaseta, I. Campbell, P. Torreri, S. Hoos, P. England, Y. Liu, Y. Abdiche, D. Malashock, A. Pinkerton, M. Wong, E. Lafer, C. Hinck, K. Thompson, C.D. Primo, A. Joyce, J. Brooks, F. Torta, A.B. Bagge Hagel, J. Krarup, J. Pass, M. Ferreira, S. Shikov, M. Mikolajczyk, Y. Abe, G. Barbato, A.M. Giannetti, G. Krishnamoorthy, B. Beusink, D. Satpaev, T. Tsang, E. Fang, J. Partridge, S. Brohawn, J. Horn, O. Pritsch, G. Obal, S. Nilapwar, B. Busby, G. Gutierrez-Sanchez, R.D. Gupta, S. Canepa, K. Witte, Z. Nikolovska-Coleska, Y.H. Cho, R. D’Agata, K. Schlick, R. Calvert, E.M. Munoz, M.J. Hernaiz, T. Bravman, M. Dines, M.H. Yang, D.G. Myszka, Anal. Biochem. 386, 194 (2009)CrossRefGoogle Scholar
  3. 3.
    D. Myszka, X. He, M. Dembo, T. Morton, B. Goldstein, Biophys. J. 75, 583 (1998)CrossRefGoogle Scholar
  4. 4.
    S.P. Yadav, S. Bergqvist, M.L. Doyle, T.A. Neubert, A.P. Yamniuk, J. Biomol. Tech. 23, 94 (2012)CrossRefGoogle Scholar
  5. 5.
    A. Onell, K. Andersson, J. Mol. Recognit. 18, 307 (2005)CrossRefGoogle Scholar
  6. 6.
    Y.N. Abdiche, Biosensors 302, 81 (2010)Google Scholar
  7. 7.
    D.G. Myszka, J. Mol. Recognit. 12, 279 (1999)CrossRefGoogle Scholar
  8. 8.
    D.G. Myszka, M.D. Jonsen, B.J. Graves, Anal. Biochem. 265, 326 (1998)CrossRefGoogle Scholar
  9. 9.
    R. Barbour, M.P. Bova, Bioanalysis 4, 619 (2012)CrossRefGoogle Scholar
  10. 10.
    I. Navratilova, E. Eisenstien, D.G. Myszka, Anal. Biochem. 344, 295 (2005)CrossRefGoogle Scholar
  11. 11.
    B. Ma, M. Alam, E. Go, X. Lu, H. Desaire, G. Tomaras, C. Bowman, L. Sutherland, R. Scearce, S. Santra, N. Letvin, T. Kepler, H. Liao, B. Haynes, Plos Pathogens 7, 1 (2011)CrossRefGoogle Scholar
  12. 12.
    T. Do, F. Ho, B. Heidecker, K. Witte, L. Chang, L. Lerner, Protein Expr. Purif. 60, 150 (2008)CrossRefGoogle Scholar
  13. 13.
    D.J. O’Shannessy, M. Brigham-Burke, K.K. Soneson, P. Hensley, I. Brooks, Anal. Biochem. 212, 457 (1993)CrossRefGoogle Scholar
  14. 14.
    J. Tintner, M. Kühleitner, E. Binner, N. Brunner, E. Smidt, Biodegradation 23, 407 (2012)CrossRefGoogle Scholar
  15. 15.
    A. Holmberg, A. Blomstergren, O. Nord, M. Lukacs, J. Lundeberg, M. Uhlen, Electrophoresis 26, 501 (2005)CrossRefGoogle Scholar
  16. 16.
    Y.N. Abdiche, D.G. Myszka, Anal. Biochem. 328, 233 (2004)CrossRefGoogle Scholar
  17. 17.
    J.R. Mascola, M.K. Louder, T.C. VanCott, C.V. Sapan, J.S. Lambert, L.R. Muenz, B. Bunow, D.L. Birx, M.L. Robb, J. Virol. 71, 7198 (1997)Google Scholar
  18. 18.
    A. Dey, K. David, P. Klasse, J. Moore, Virology 360, 199 (2007)CrossRefGoogle Scholar
  19. 19.
    Y. Abdiche, D. Malashock, A. Pinkerton, J. Pons, Anal. Biochem. 377, 209 (2008)CrossRefGoogle Scholar
  20. 20.
    J. Wallner, G. Lhota, D. Jeschek, A. Mader, K. Vorauer-Uhl, J. Pharm. Biomed. Anal. 72, 150 (2013)CrossRefGoogle Scholar
  21. 21.
    P.S. Katsamba, I. Navratilova, M. Calderon-Cacia, L. Fan, K. Thornton, M. Zhu, T.V. Bos, C. Forte, D. Friend, I. Laird-Offringa, G. Tavares, J. Whatley, E. Shi, A. Widom, K.C. Lindquist, S. Klakamp, A. Drake, D. Bohmann, M. Roell, L. Rose, J. Dorocke, B. Roth, B. Luginbühl, D.G. Myszka, Anal. Biochem. 352, 208 (2006)CrossRefGoogle Scholar
  22. 22.
    D.J. O’Shannessy, M. Brigham-Burke, K.K. Soneson, P. Hensley, I. Brooks, Methods Enzymol. 240, 323 (1994)Google Scholar

Copyright information

© Springer Science+Business Media New York 2013

Authors and Affiliations

  • Jakob Wallner
    • 1
    Email author
  • Manfred Kühleitner
    • 2
  • Norbert Brunner
    • 2
  • Gabriele Lhota
    • 1
  • Karola Vorauer-Uhl
    • 1
  1. 1.Department of BiotechnologyUniversity of Natural Resources and Life SciencesViennaAustria
  2. 2.Department of Integrative Biology, Institute of MathematicsUniversity of Natural Resources and Life SciencesViennaAustria

Personalised recommendations