Isolation of Llama Antibody Fragments for Prevention of Dandruff by Phage Display in Shampoo

• Published in: Applied and Environmental Microbiology Vol. 71; no. 1; pp. 442 - 450
• Main Authors: Dolk, Edward, van der Vaart, Marcel, Lutje Hulsik, David, Vriend, Gert, de Haard, Hans, Spinelli, Silvia, Cambillau, Christian, Frenken, Leon, Verrips, Theo
• Format: Journal Article
• Language: English
• Published: Washington, DC American Society for Microbiology 2005
• Subjects: Amino Acid Sequence; Amino acids; Animals; Antibodies; Antibodies, Fungal - chemistry; Antibodies, Fungal - genetics; Antibodies, Fungal - immunology; Antibodies, Fungal - isolation & purification; Antibody Specificity; Arginine; Binding sites; Biological and medical sciences; Biotechnology; Camelids, New World - immunology; Cell surface; Cell walls; Dermatitis, Seborrheic - microbiology; Dermatitis, Seborrheic - prevention & control; Dermatomycoses - microbiology; Dermatomycoses - prevention & control; Feasibility studies; Fundamental and applied biological sciences. Psychology; Fungal Proteins - administration & dosage; Fungal Proteins - immunology; Fungi; Hair Preparations; Immunoglobulin Fragments - chemistry; Immunoglobulin Fragments - genetics; Immunoglobulin Fragments - immunology; Immunoglobulin Fragments - isolation & purification; Immunoglobulin Heavy Chains - chemistry; Immunoglobulin Heavy Chains - genetics; Immunoglobulin Heavy Chains - immunology; Immunoglobulin Variable Region - chemistry; Immunoglobulin Variable Region - genetics; Immunoglobulin Variable Region - immunology; Malassezia - immunology; Malassezia furfur; Molecular Sequence Data; Organisms; Peptide Library; Phage display; Physical growth; Protein engineering; Public Health Microbiology; Q1; Q2; Scalp Dermatoses - microbiology; Scalp Dermatoses - prevention & control; Shampoos; Surfactants; Urea; Fungi; Malassezia furfur; Prevention; Phage display; Yeast; Antibody; Shampoo; Isolation; Fungi Imperfecti; Dandruff; Thallophyta
• ISSN: 0099-2240; 1098-5336
• DOI: 10.1128/aem.71.1.442-450.2005

As part of research exploring the feasibility of using antibody fragments to inhibit the growth of organisms implicated in dandruff, we isolated antibody fragments that bind to a cell surface protein of Malassezia furfur in the presence of shampoo. We found that phage display of llama single-domain antibody fragments (VHHs) can be extended to very harsh conditions, such as the presence of shampoo containing nonionic and anionic surfactants. We selected several VHHs that bind to the cell wall protein Malf1 of M. furfur, a fungus implicated in causing dandruff. In addition to high stability in the presence of shampoo, these VHHs are also stable under other denaturing conditions, such as high urea concentrations. Many of the stable VHHs were found to contain arginine at position 44. Replacement of the native amino acid at position 44 with arginine in the most stable VHH that lacked this arginine resulted in a dramatic further increase in the stability. The combination of the unique properties of VHHs together with applied phage display and protein engineering is a powerful method for obtaining highly stable VHHs that can be used in a wide range of applications.