Sequence analysis of feline immunoglobulin mRNAs and the development of a felinized monoclonal antibody specific to feline panleukopenia virus
| Autor: | Zhengchun Lu, John S. L. Parker, M. Julia B. Felippe, Rebecca L. Tallmadge, Heather M. Callaway |
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| Rok vydání: | 2017 |
| Předmět: |
0301 basic medicine
Feline Panleukopenia medicine.drug_class Sequence analysis lcsh:Medicine Feline panleukopenia Antibodies Viral Monoclonal antibody Immunoglobulin light chain Peripheral blood mononuclear cell Article Virus Immunoglobulin kappa-Chains 03 medical and health sciences Immunoglobulin lambda-Chains medicine Animals RNA Messenger lcsh:Science B-Lymphocytes Multidisciplinary biology Sequence Analysis RNA lcsh:R Antibodies Monoclonal biology.organism_classification Virology Molecular biology Immunoglobulin A 030104 developmental biology Immunoglobulin G Cats biology.protein lcsh:Q Feline Panleukopenia Virus Antibody Immunoglobulin Heavy Chains Clone (B-cell biology) |
| Zdroj: | Scientific Reports Scientific Reports, Vol 7, Iss 1, Pp 1-15 (2017) |
| ISSN: | 2045-2322 |
| DOI: | 10.1038/s41598-017-12725-5 |
| Popis: | In response to immunization, B-cells generate a repertoire of antigen-specific antibodies. Antibody-based immunotherapies hold great promise for treating a variety of diseases in humans. Application of antibody-based immunotherapy in cats is limited by the lack of species-specific complete sequences for mRNAs encoding rearranged heavy and light chain immunoglobulins in B cells. To address this barrier, we isolated mRNAs from feline peripheral blood mononuclear cells (PBMCs), and used available immunoglobulin sequences and 5′ and 3′ RACE to clone and sequence heavy and light chain immunoglobulin mRNAs. We recovered mRNA from PBMCs from two cats, cloned and sequenced the variable and constant domains of the feline heavy chains of IgG1a (IGHG1a), IgG2 (IGHG2), and IgA (IGHA), and the light chains (lambda and kappa). Using these sequences, we prepared two bicistronic vectors for mammalian expression of a representative feline heavy (IGHG1a) together with a light (lambda or kappa) chain. Here we report novel feline Ig sequences, a technique to express antigen-specific felinized monoclonal antibodies, and the initial characterization of a functional felinized monoclonal antibody against feline panleukopenia virus. |
| Databáze: | OpenAIRE |
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