Human IgA Monoclonal Antibodies That Neutralize Poliovirus, Produced by Hybridomas and Recombinant Expression

Autor: Diana Kouiavskaia, Fetweh H. Al-Saleem, Chandana Devi Kattala, Konstantin Chumakov, Rama Devudu Puligedda, Jiang-Yang Zhao, Vladimir Vigdorovich, Scott K. Dessain, D. Noah Sather
Rok vydání: 2020
Předmět:
Zdroj: Antibodies
Volume 9
Issue 1
Antibodies, Vol 9, Iss 1, p 5 (2020)
ISSN: 2073-4468
Popis: Poliovirus (PV)-specific intestinal IgAs are important for cessation of PV shedding in the gastrointestinal tract following an acute infection with wild type or vaccine-derived PV strains. We sought to produce IgA monoclonal antibodies (mAbs) with PV neutralizing activity. We first performed de novo IgA discovery from primary human B cells using a hybridoma method that allows assessment of mAb binding and expression on the hybridoma surface: On-Cell mAb Screening (OCMS&trade
). Six IgA1 mAbs were cloned by this method
three potently neutralized type 3 Sabin and wt PV strains. The hybridoma mAbs were heterogeneous, expressed in monomeric, dimeric, and aberrant forms. We also used recombinant methods to convert two high-potency anti-PV IgG mAbs into dimeric IgA1 and IgA2 mAbs. Isotype switching did not substantially change their neutralization activities. To purify the recombinant mAbs, Protein L binding was used, and one of the mAbs required a single amino acid substitution in its &kappa
LC in order to enable protein L binding. Lastly, we used OCMS to assess IgA expression on the surface of hybridomas and transiently transfected, adherent cells. These studies have generated potent anti-PV IgA mAbs, for use in animal models, as well as additional tools for the discovery and production of human IgA mAbs.
Databáze: OpenAIRE
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