AAPS PharmSci

, 5:1 | Cite as

Development of a multidose formulation for a humanized monoclonal antibody using experimental design techniques



The purpose of this study was to identify optimal preservatives for a multidose formulation of a humanized monoclonal antibody using experimental design techniques. The effect of antimicrobial parenteral preservatives (benzyl alcohol, chlorobutanol, methyl paraben, propylparaben, phenol, and m-cresol) on protein stability was assessed using size-exclusion chromatography, differential scanning calorimetry, right-angle light scattering, UV spectroscopy, and potency testing using a cell-based fluorescence-activated cell sorting method. A quick, cost-effective preservative screening test was designed. Combinations of preservatives were examined using an I-optimal experimental design. The protein was most stable in the presence of methylparaben and propylparaben, and was compatible with benzyl alcohol and chlorobutanol at low concentrations. Phenol and m-cresol were not compatible with the protein. The I-optimal experimental design indicated that as an individual preservative, benzyl alcohol was promising. The model also indicated several effective combinations of preservatives that satisfied the antimicrobial efficacy and physical stability constraints. The preservative screening test and the experimental design approach were effective in identifying optimal concentrations of antimicrobial preservatives for a multidose protein formulation; (1) benzyl alcohol, and (2) the combination of methylparaben and chlorobutanol were screened as potential candidates to satisfy the regulatory requirements of various preservative efficacy tests.


multidose formulation preservative experimental design monoclonal antibody protein 


  1. 1.
    McGoff P, Scher DS. Solution formulation of proteins/peptides. In: McNally EJ, ed. Protein Formulation and Delivery. New York, NY: Marcel Dekker, 2000:139–158.Google Scholar
  2. 2.
    Akers MJ, Defelippis MR. Peptides and proteins as parenteral solutions. In: Frokjaer S, Hovgaard L, eds. Pharmaceutical Formulation Development of Peptides and Proteins. Philadelphia, PA: Taylor and Francis; 2000:145–177.Google Scholar
  3. 3.
    Bontempo JA. Formulation development. In: Bontempo JA, ed. Development of Biopharmaceutical Parenteral Dosage Forms. New York, NY: Marcel Dekker; 1997:109–142.Google Scholar
  4. 4.
    Lachman L, Weinstein S, Hopkins G, Slack S, Eisman P, Cooper J. Stability of antibacterial preservatives in parenteral solutions, I. Factors Influencing the Loss of Antimicrobial Agents from Solutions in Rubber-Stoppered Containers, J Pharm Sci. 1962;51(3):224–232.PubMedCrossRefGoogle Scholar
  5. 5.
    Kakemi K, Sezaki H, Arakawa E, Ideda K. Interaction of parabens and other pharmaceutical adjuvants with plastic containers. J Chem Pharm Bull. 1971;19(12):2523–2529.CrossRefGoogle Scholar
  6. 6.
    Akers MJ. Considerations in selecting antimicrobial preservatives for parenteral product development. Pharm Technol. 1984;8(5):36–44.Google Scholar
  7. 7.
    Beasley CRW, Rafferty P, Holgate ST. Bronchoconstrictor properties of preservatives in ipratropium bromide (Atrovent) nebulizer solution. Br Med J. 1987;294:1197–1198.CrossRefGoogle Scholar
  8. 8.
    Maa YF, Hsu CC. Aggregation of recombinant human growth hormone induced by phenolic compounds. Int J Pharm. 1996;140:155–168.CrossRefGoogle Scholar
  9. 9.
    Kim Y, Rose CA, Liu Y, Ozaki Y, Datta G, Ut AT. FT-IR and near-infrared FT-Raman studies of the secondary structure of insulinotropin in the solid state: α-helix to β-sheet conversion by phenol and/or high shear force. J Pharm Sci. 1994;83:1175–1180.PubMedCrossRefGoogle Scholar
  10. 10.
    Lam XM, Patapoff TW, Nguyen TH. The effect of benzyl alcohol on recombinant human interferon-γ. Pharm Res. 1997;14:725–729.PubMedCrossRefGoogle Scholar

Copyright information

© American Association of Pharmaceutical Scientists 2003

Authors and Affiliations

  1. 1.Analytical and Formulation SciencesProtein Design Labs, IncFremont

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