Cover Image

When monoclonal antibodies are not monospecific: Hybridomas frequently express additional functional variable regions

Monoclonal antibodies are commonly assumed to be monospecific, but anecdotal studies have reported genetic diversity in antibody heavy chain and light chain genes found within individual hybridomas. As the prevalence of such diversity has never been explored, we analyzed 185 random hybridomas, in a...

Full description

Saved in:
Bibliographic Details
Main Authors: Bradbury, Andrew (Author) , Trinklein, Nathan D. (Author) , Thie, Holger (Author) , Wilkinson, Ian C. (Author) , Tandon, Atul K. (Author) , Anderson, Stephen (Author) , Bladen, Catherine L. (Author) , Jones, Brittany (Author) , Aldred, Shelley Force (Author) , Bestagno, Marco (Author) , Burrone, Oscar (Author) , Maynard, Jennifer (Author) , Ferrara, Fortunato (Author) , Trimmer, James S. (Author) , Görnemann, Janina (Author) , Glanville, Jacob (Author) , Wolf, Philipp (Author) , Frenzel, Andre (Author) , Wong, Julin (Author) , Koh, Xin Yu (Author) , Eng, Hui-Yan (Author) , Lane, David (Author) , Lefranc, Marie-Paule (Author) , Clark, Mike (Author) , Dübel, Stefan (Author)
Format: Article (Journal)
Language:English
Published: 29 Mar 2018
In: mAbs
Year: 2018, Volume: 10, Issue: 4, Pages: 539-546
ISSN:1942-0870
DOI:10.1080/19420862.2018.1445456
Online Access:Verlag, lizenzpflichtig, Volltext: https://doi.org/10.1080/19420862.2018.1445456
Get full text
Author Notes:Andrew R.M. Bradbury, Nathan D. Trinklein, Holger Thie, Ian C. Wilkinson, Atul K. Tandon, Stephen Anderson, Catherine L. Bladen, Brittany Jones, Shelley Force Aldred, Marco Bestagno, Oscar Burrone, Jennifer Maynard, Fortunato Ferrara, James S. Trimmer, Janina Görnemann, Jacob Glanville, Philipp Wolf, Andre Frenzel, Julin Wong, Xin Yu Koh, Hui-Yan Eng, David Lane, Marie-Paule Lefranc, Mike Clark & Stefan Dübel
Description
Summary:Monoclonal antibodies are commonly assumed to be monospecific, but anecdotal studies have reported genetic diversity in antibody heavy chain and light chain genes found within individual hybridomas. As the prevalence of such diversity has never been explored, we analyzed 185 random hybridomas, in a large multicenter dataset. The hybridomas analyzed were not biased towards those with cloning difficulties or known to have additional chains. Of the hybridomas we evaluated, 126 (68.1%) contained no additional productive chains, while the remaining 59 (31.9%) contained one or more additional productive heavy or light chains. The expression of additional chains degraded properties of the antibodies, including specificity, binding signal and/or signal-to-noise ratio, as determined by enzyme-linked immunosorbent assay and immunohistochemistry. The most abundant mRNA transcripts found in a hybridoma cell line did not necessarily encode the antibody chains providing the correct specificity. Consequently, when cloning antibody genes, functional validation of all possible VH and VL combinations is required to identify those with the highest affinity and lowest cross-reactivity. These findings, reflecting the current state of hybridomas used in research, reiterate the importance of using sequence-defined recombinant antibodies for research or diagnostic use.
Item Description:Gesehen am 02.12.2020
Physical Description:Online Resource
ISSN:1942-0870
DOI:10.1080/19420862.2018.1445456

Similar Items