tetano
Editor, Senior Moderator
Biologicals. 2016 Jul 24. pii: S1045-1056(16)30031-8. doi: 10.1016/j.biologicals.2016.05.006. [Epub ahead of print]
[h=1]Recombinant production and characterization of human anti-influenza virus monoclonal antibodies identified from hybridomas fused with human lymphocytes.[/h] Misaki R[SUP]1[/SUP], Fukura N[SUP]2[/SUP], Kajiura H[SUP]2[/SUP], Yasugi M[SUP]3[/SUP], Kubota-Koketsu R[SUP]4[/SUP], Sasaki T[SUP]5[/SUP], Momota M[SUP]6[/SUP], Ono KI[SUP]7[/SUP], Ohashi T[SUP]2[/SUP], Ikuta K[SUP]5[/SUP], Fujiyama K[SUP]8[/SUP].
[h=3]Author information[/h]
[h=3]Abstract[/h] 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.
[h=4]KEYWORDS:[/h] Chinese hamster ovary; Glycosylation; Human antibody production; SPYMEG
PMID: 27464991 DOI: 10.1016/j.biologicals.2016.05.006
[PubMed - as supplied by publisher]
[h=1]Recombinant production and characterization of human anti-influenza virus monoclonal antibodies identified from hybridomas fused with human lymphocytes.[/h] Misaki R[SUP]1[/SUP], Fukura N[SUP]2[/SUP], Kajiura H[SUP]2[/SUP], Yasugi M[SUP]3[/SUP], Kubota-Koketsu R[SUP]4[/SUP], Sasaki T[SUP]5[/SUP], Momota M[SUP]6[/SUP], Ono KI[SUP]7[/SUP], Ohashi T[SUP]2[/SUP], Ikuta K[SUP]5[/SUP], Fujiyama K[SUP]8[/SUP].
[h=3]Author information[/h]
[h=3]Abstract[/h] 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.
[h=4]KEYWORDS:[/h] Chinese hamster ovary; Glycosylation; Human antibody production; SPYMEG
PMID: 27464991 DOI: 10.1016/j.biologicals.2016.05.006
[PubMed - as supplied by publisher]