Enhancement of toxin- and virus-neutralizing capacity of single-domain antibody fragments by N-glycosylation
| Autor: | M.M. Harmsen, C. B. van Solt, H.P.D. Fijten |
|---|---|
| Rok vydání: | 2009 |
| Předmět: |
binding
Glycosylation N-glycosylation medicine.disease_cause Applied Microbiology and Biotechnology Epitope chemistry.chemical_compound Enterotoxins Epitopes N-linked glycosylation Chlorocebus aethiops Biotechnologically Relevant Enzymes and Proteins Immunoglobulin Fragments Escherichia coli Proteins Bacteriologie Cholera toxin Bacteriology Host Pathogen Interaction & Diagnostics General Medicine Virology & Molecular Biology secretion Foot-and-Mouth Disease Virus saccharomyces-cerevisiae lipids (amino acids peptides and proteins) vivo Camelids New World Biotechnology cholera-toxin Recombinant Fusion Proteins monoclonal-antibodies Bacterial Toxins Molecular Sequence Data macromolecular substances Saccharomyces cerevisiae in-vitro Biology Heat-labile enterotoxin Neutralization expression medicine Animals Amino Acid Sequence Vero Cells heat-labile enterotoxin Host Pathogen Interaction & Diagnostics Bacteriology Molecular biology Fusion protein Antibodies Neutralizing Host Pathogen Interactie & Diagnostiek proteins Yeast Virologie & Moleculaire Biologie carbohydrates (lipids) Single-domain antibody chemistry Recombinant antibody Foot-and-Mouth Disease Bacteriologie Host Pathogen Interactie & Diagnostiek Nanobody |
| Zdroj: | Applied Microbiology and Biotechnology Applied Microbiology and Biotechnology 84 (2009) 6 Applied Microbiology and Biotechnology, 84(6), 1087-1094 |
| ISSN: | 1432-0614 0175-7598 |
| Popis: | Single-domain antibody fragments (VHHs) have several beneficial properties as compared to conventional antibody fragments. However, their small size complicates their toxin- and virus-neutralizing capacity. We isolated 27 VHHs binding Escherichia coli heat-labile toxin and expressed these in Saccharomyces cerevisiae. The most potent neutralizing VHH (LT109) was N-glycosylated, resulting in a large increase in molecular mass. This suggests that N-glycosylation of LT109 improves its neutralizing capacity. Indeed, deglycosylation of LT109 decreased its neutralizing capacity three- to fivefold. We also studied the effect of glycosylation of two previously isolated VHHs on their ability to neutralize foot-and-mouth disease virus. For this purpose, these VHHs that lacked potential N-glycosylation sites were genetically fused to another VHH that was known to be glycosylated. The resulting fusion proteins were also N-glycosylated. They neutralized the virus at at least fourfold-lower VHH concentrations as compared to the single, non-glycosylated VHHs and at at least 50-fold-lower VHH concentrations as compared to their deglycosylated counterparts. Thus, we have shown that N-glycosylation of VHHs contributes to toxin- and virus-neutralizing capacity. |
| Databáze: | OpenAIRE |
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