Open AccessComparison of VHH ‐Fc antibody production in A rabidopsis thaliana , N icotiana benthamiana and P ichia pastoris
Author(s) -
De Meyer Thomas,
Laukens Bram,
Nolf Jonah,
Van Lerberge Els,
De Rycke Riet,
De Beuckelaer Ans,
De Buck Sylvie,
Callewaert Nico,
Depicker Ann
Publication year - 2015
Publication title -
plant biotechnology journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.525
H-Index - 115
eISSN - 1467-7652
pISSN - 1467-7644
DOI - 10.1111/pbi.12330
Subject(s) - nicotiana benthamiana , kdel , biology , microbiology and biotechnology , glycosylation , endoplasmic reticulum , biochemistry , golgi apparatus , gene
Summary VHH s or nanobodies are widely acknowledged as interesting diagnostic and therapeutic tools. However, for some applications, multivalent antibody formats, such as the dimeric VHH ‐Fc format, are desired to increase the functional affinity. The scope of this study was to compare transient expression of diagnostic VHH ‐Fc antibodies in N icotiana benthamiana leaves with their stable expression in A rabidopsis thaliana seeds and P ichia pastoris . To this end, VHH ‐Fc antibodies targeting green fluorescent protein or the A . thaliana seed storage proteins (albumin and globulin) were produced in the three platforms. Differences were mainly observed in the accumulation levels and glycosylation patterns. Interestingly, although in plants oligomannosidic N‐glycans were expected for KDEL ‐tagged VHH ‐Fcs, several VHH ‐Fcs with an intact KDEL ‐tag carried complex‐type N‐glycans, suggesting a dysfunctional retention in the endoplasmic reticulum. All VHH ‐Fcs were equally functional across expression platforms and several outperformed their corresponding VHH in terms of sensitivity in ELISA .