Production of Monoclonal Antibody That Recognizes Zika Virus and Other Flaviviruses in Serum-Free Conditions
| Autor: | Raquel de Souza Martins, Sotiris Missailidis, Sheila Maria Barbosa de Lima, Patricia Barbosa Jurgilas, Hilton Jorge Nascimento, Ana Carolina dos Reis Albuquerque Cajaraville, Márcia Arissawa, Fernando de Paiva Conte |
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| Rok vydání: | 2017 |
| Předmět: |
0301 basic medicine
medicine.drug_class Immunology Dengue virus medicine.disease_cause Monoclonal antibody Culture Media Serum-Free Zika virus 03 medical and health sciences Epitopes 0302 clinical medicine Antigen Chlorocebus aethiops medicine Immunology and Allergy Animals 030212 general & internal medicine Vero Cells Mice Inbred BALB C Hybridomas biology Flavivirus Antibodies Monoclonal Zika Virus biology.organism_classification Virology 030104 developmental biology Batch Cell Culture Techniques Monoclonal biology.protein Electrophoresis Polyacrylamide Gel Antibody Fetal bovine serum |
| Zdroj: | Monoclonal antibodies in immunodiagnosis and immunotherapy. 36(6) |
| ISSN: | 2167-9436 |
| Popis: | With the recent outbreaks of Zika and Dengue virus infections in various countries worldwide, production of vaccines or diagnostic kits is an urgent public health demand. Production of a monoclonal antibody (mAb) that specifically binds to a common antigen shared by the Flavivirus genus will be necessary for new diagnostic kits or characterization and viral identity tests during vaccine development. This study aimed to cultivate, in serum-free conditions, the 4G2 hybridoma that produces an mAb, which recognizes a shared epitope from the Flavivirus genus. We compared 4G2 hybridoma growth and biochemical profiles between cells cultivated in batch mode over 10 days in roller bottles containing Dulbecco's modified Eagle's medium high glucose containing 10% fetal bovine serum medium or hybridomas directly adapted to Ex-Cell serum-free medium. Cellular parameters such as specific growth rate (μ), maximum cell concentration, specific l-lactate, and glucose and IgG rates were evaluated. Thereafter, we also compared total mAb volumetric productivity, purification yield, and mAb staining of Vero cells infected with Zika and Dengue-2 virus. Direct adaptation to serum-free conditions did not change hybridoma growth rate and mAb production under the conditions tested. Instead, serum-free mAb purification showed a higher yield with no alterations on mAb structure or mAb staining of Zika and Dengue Vero-infected cells. |
| Databáze: | OpenAIRE |
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