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Generation and Application of Monoclonal Antibody Against Lycopene.
- Source :
-
Monoclonal antibodies in immunodiagnosis and immunotherapy [Monoclon Antib Immunodiagn Immunother] 2017 Apr; Vol. 36 (2), pp. 62-67. Date of Electronic Publication: 2017 Apr 12. - Publication Year :
- 2017
-
Abstract
- A monoclonal antibody (Mab) against lycopene was developed from hybridoma clones obtained from BALB/c mice immunized with trans-isomer of lycopene (t-lycopene, t-LC) conjugated with colloidal gold particles. An alternating immunization schedule which included injection of both formulations of immunogen (without and with Freund's adjuvant) was most effective in the elucidation of a measurable immune response to the t-Lycopene conjugate. Selected hybridoma clones were able to produce an Mab positive in competition assay. In particular, preincubation of 6B9 Mabs with t-LC abolished the ability of 6B9 Mabs to bind LC in the competition assay. Mabs produced by other clones (4F10, 4A3, and 3B12) worked similarly. Analysis of antigen specificity showed that 6B9 Mab raised against t-LC did not recognize other carotenoids such as lutein and carotene. Mab 6B9 was shown to recognize lycopene on a glass surface and in the settings of indirect immunofluorescence experiments performed in cultured hepatocytes and alveolar macrophages incubated with and without lycopene, as well as in sebum and corneocyte specimens from the skin of volunteers supplemented with nutraceutical formulation of lycopene. Newly generated Mabs against lycopene may provide a valuable tool for different analytical assays of lycopene content in various biological, agricultural, and food products.
- Subjects :
- Animals
Antibodies, Monoclonal biosynthesis
Antibodies, Monoclonal chemistry
Antibody Specificity
Antigens administration & dosage
Antigens chemistry
Blotting, Western
Carotenoids administration & dosage
Carotenoids chemistry
Cells, Cultured
Enzyme-Linked Immunosorbent Assay
Fluorescent Antibody Technique, Indirect
Freund's Adjuvant administration & dosage
Gold Colloid administration & dosage
Gold Colloid chemistry
Hepatocytes chemistry
Hepatocytes ultrastructure
Humans
Hybridomas immunology
Immunoconjugates administration & dosage
Immunoconjugates chemistry
Lutein
Lycopene
Macrophages, Alveolar chemistry
Macrophages, Alveolar ultrastructure
Mice
Mice, Inbred BALB C
Spleen cytology
Spleen immunology
Antibodies, Monoclonal isolation & purification
Antigens immunology
Carotenoids immunology
Immunization, Secondary methods
Immunoconjugates immunology
Subjects
Details
- Language :
- English
- ISSN :
- 2167-9436
- Volume :
- 36
- Issue :
- 2
- Database :
- MEDLINE
- Journal :
- Monoclonal antibodies in immunodiagnosis and immunotherapy
- Publication Type :
- Academic Journal
- Accession number :
- 28402743
- Full Text :
- https://doi.org/10.1089/mab.2016.0046