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Biologicals. 2016 Jul 24. pii: S1045-1056(16)30031-8. doi: 10.1016/j.biologicals.2016.05.006. [Epub ahead of print]
Recombinant production and characterization of human anti-influenza virus monoclonal antibodies identified from hybridomas fused with human lymphocytes.
Misaki R1, Fukura N2, Kajiura H2, Yasugi M3, Kubota-Koketsu R4, Sasaki T5, Momota M6, Ono KI7, Ohashi T2, Ikuta K5, Fujiyama K8.
Author information
Abstract
In previous studies, hybridomas producing human immunoglobulin G, the antibodies 5E4 and 5A7 against influenza A and B virus were established using a novel human lymphocyte fusion partner, SPYMEG. In the present study, we succeeded in achieving the recombinant production and secretion of 5E4 and 5A7 in Chinese hamster ovary cells. Our N-glycan analysis by intact-mass detection and liquid chromatography mass spectrometry showed that recombinant 5E4 and 5A7 have one N-glycan and the typical mammalian-type N-glycan structures similar to those in hybridomas. However, the glycan distribution was slightly different among these antibodies. The amount of high-mannose-type structures was under 10% of the total N-glycans of recombinant 5E4 and 5A7, compared to 20% of the 5E4 and 5A7 produced in hybridomas. The amount of galactosylated N-glycans was increased in recombinants. Approximately 80% of the N-glycans of all antibodies was fucosylated, and no sialylated N-glycan was found. Recombinant 5E4 and 5A7 neutralized pandemic influenza A virus specifically, and influenza B virus broadly, quite similar to the 5E4 and 5A7 produced in hybridomas, respectively. Here we demonstrated that recombinants of antibodies identified from hybridomas fused with SPYMEG have normal N-glycans and that their neutralizing activities bear comparison with those of the original antibodies.
Copyright ? 2016 International Alliance for Biological Standardization. Published by Elsevier Ltd. All rights reserved.
KEYWORDS:
Chinese hamster ovary; Glycosylation; Human antibody production; SPYMEG
PMID: 27464991 DOI: 10.1016/j.biologicals.2016.05.006
[PubMed - as supplied by publisher]
Recombinant production and characterization of human anti-influenza virus monoclonal antibodies identified from hybridomas fused with human lymphocytes.
Misaki R1, Fukura N2, Kajiura H2, Yasugi M3, Kubota-Koketsu R4, Sasaki T5, Momota M6, Ono KI7, Ohashi T2, Ikuta K5, Fujiyama K8.
Author information
Abstract
In previous studies, hybridomas producing human immunoglobulin G, the antibodies 5E4 and 5A7 against influenza A and B virus were established using a novel human lymphocyte fusion partner, SPYMEG. In the present study, we succeeded in achieving the recombinant production and secretion of 5E4 and 5A7 in Chinese hamster ovary cells. Our N-glycan analysis by intact-mass detection and liquid chromatography mass spectrometry showed that recombinant 5E4 and 5A7 have one N-glycan and the typical mammalian-type N-glycan structures similar to those in hybridomas. However, the glycan distribution was slightly different among these antibodies. The amount of high-mannose-type structures was under 10% of the total N-glycans of recombinant 5E4 and 5A7, compared to 20% of the 5E4 and 5A7 produced in hybridomas. The amount of galactosylated N-glycans was increased in recombinants. Approximately 80% of the N-glycans of all antibodies was fucosylated, and no sialylated N-glycan was found. Recombinant 5E4 and 5A7 neutralized pandemic influenza A virus specifically, and influenza B virus broadly, quite similar to the 5E4 and 5A7 produced in hybridomas, respectively. Here we demonstrated that recombinants of antibodies identified from hybridomas fused with SPYMEG have normal N-glycans and that their neutralizing activities bear comparison with those of the original antibodies.
Copyright ? 2016 International Alliance for Biological Standardization. Published by Elsevier Ltd. All rights reserved.
KEYWORDS:
Chinese hamster ovary; Glycosylation; Human antibody production; SPYMEG
PMID: 27464991 DOI: 10.1016/j.biologicals.2016.05.006
[PubMed - as supplied by publisher]