1. Reduced immunogenicity of beta-lactoglobulin by conjugating with chitosan
- Author
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Tomomi Aoki, Koji Takahashi, Makoto Hattori, Tadashi Yoshida, and Silvia Iskandar
- Subjects
Antigenicity ,medicine.drug_class ,Enzyme-Linked Immunosorbent Assay ,Lactoglobulins ,Monoclonal antibody ,Applied Microbiology and Biotechnology ,Biochemistry ,Epitope ,Analytical Chemistry ,chemistry.chemical_compound ,Mice ,medicine ,Animals ,Vitamin A ,Molecular Biology ,Carbodiimide ,Chitosan ,Mice, Inbred C3H ,integumentary system ,Linear epitope ,Molecular Structure ,Chemistry ,Immunogenicity ,Organic Chemistry ,General Medicine ,Molecular biology ,Carbodiimides ,Isoelectric point ,Cross-Linking Reagents ,Epitopes, B-Lymphocyte ,Cattle ,Biotechnology ,Conjugate - Abstract
Bovine beta-lactoglobulin (beta-LG) was conjugated with chitosan (CHS) by means of a water-soluble carbodiimide to reduce the immunogenicity of beta-LG. Each beta-LG-CHS conjugate was purified by ion-exchange chromatography and hydrophobic chromatography. The conjugation between beta-LG and CHS was confirmed by SDS-PAGE, the isoelectric point of the conjugate being higher than that of beta-LG. Two types of the beta-LG-CHS conjugate were obtained with molar ratios of beta-LG to CHS of 1:1 (F1) and 1:2 (F2). Structural analyses by fluorescence measurement, ELISA with monoclonal antibodies and retinol-binding activity indicated that the conjugates had almost maintained the native structure of beta-LG. The antigenicity of the beta-LG-CHS conjugates was similar to that of beta-LG in C3H/He mice. Reduction of the immunogenicity of beta-LG was achieved by conjugation with CHS. In particular, F2 showed very low immunogenicity. B cell epitopes of beta-LG and the conjugates recognized in C3H/He mice were determined with 15-mer multi-pin peptide; the linear epitope profiles of the conjugates were found to be similar to those of beta-LG, while the antibody response to each epitope was dramatically reduced. Conjugation of beta-LG with chitosan was effective for reducing the immunogenicity of beta-LG.
- Published
- 2006