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When monoclonal antibodies are not monospecific: Hybridomas frequently express additional functional variable regions.
Bradbury, Andrew R M; Trinklein, Nathan D; Thie, Holger; Wilkinson, Ian C; Tandon, Atul K; Anderson, Stephen; Bladen, Catherine L; Jones, Brittany; Aldred, Shelley Force; Bestagno, Marco; Burrone, Oscar; Maynard, Jennifer; Ferrara, Fortunato; Trimmer, James S; Görnemann, Janina; Glanville, Jacob; Wolf, Philipp; Frenzel, Andre; Wong, Julin; Koh, Xin Yu; Eng, Hui-Yan; Lane, David; Lefranc, Marie-Paule; Clark, Mike; Dübel, Stefan.
Afiliação
  • Bradbury ARM; a Specifica Inc. , 1512 Pacheco St, Santa Fe , NM , USA.
  • Trinklein ND; b SwitchGear Genomics , 1914 Palomar Oaks Way, Carlsbad , CA USA.
  • Thie H; c Miltenyi Biotec GmbH , Friedrich-Ebert-Str. 68, Bergisch Gladbach , Germany.
  • Wilkinson IC; d Absolute Antibody, Wilton Centre , Redcar , Cleveland TS10 4RF , United Kingdom.
  • Tandon AK; e NeoBiotechnologies , 2 Union Square, Union City , CA , USA.
  • Anderson S; d Absolute Antibody, Wilton Centre , Redcar , Cleveland TS10 4RF , United Kingdom.
  • Bladen CL; d Absolute Antibody, Wilton Centre , Redcar , Cleveland TS10 4RF , United Kingdom.
  • Jones B; e NeoBiotechnologies , 2 Union Square, Union City , CA , USA.
  • Aldred SF; b SwitchGear Genomics , 1914 Palomar Oaks Way, Carlsbad , CA USA.
  • Bestagno M; f International Centre for Genetic Engineering and Biotechnology (ICGEB) , Padriciano 99, Trieste , Italy.
  • Burrone O; f International Centre for Genetic Engineering and Biotechnology (ICGEB) , Padriciano 99, Trieste , Italy.
  • Maynard J; g The University of Texas at Austin, Cockrell School of Engineering , McKetta Department of Chemical Engineering , 200 E Dean Keeton St. Stop C0400, Austin , Texas , USA.
  • Ferrara F; a Specifica Inc. , 1512 Pacheco St, Santa Fe , NM , USA.
  • Trimmer JS; h Department of Physiology and Membrane Biology , University of California , Davis, One Shields Avenue, Davis , CA , USA.
  • Görnemann J; i Institute for Molecular Genetics , University of Heidelberg , Im Neuenheimer Field 260, Heidelberg , Germany.
  • Glanville J; j Stanford University, School of Medicine , Stanford , California , USA.
  • Wolf P; k Department of Urology , Medical Center, University of Freiburg , Breisacher Str. 66, Freiburg , Germany.
  • Frenzel A; l Yumab GmbH , Inhoffenstr. 7, Braunschweig , Germany.
  • Wong J; p Technische Universität Braunschweig, Institute of Biochemistry, Biotechnology and Bioinformatics , Spielmannstr. 7, Braunschweig , Germany.
  • Koh XY; m A*Star p53 laboratory , 06-06 Immunos, Singapore , Singapore.
  • Eng HY; m A*Star p53 laboratory , 06-06 Immunos, Singapore , Singapore.
  • Lane D; m A*Star p53 laboratory , 06-06 Immunos, Singapore , Singapore.
  • Lefranc MP; m A*Star p53 laboratory , 06-06 Immunos, Singapore , Singapore.
  • Clark M; n IMGT®, the international ImMunoGeneTics information system®, Laboratoire d'ImmunoGénétique Moléculaire LIGM, Institut de Génétique Humaine IGH, UPR CNRS 1142, Montpellier University , Montpellier cedex 5 , France.
  • Dübel S; o Clark Antibodies Ltd , 10 Wellington Street, Cambridge , CB1 1HW , United Kingdom.
MAbs ; 10(4): 539-546, 2018.
Article em En | MEDLINE | ID: mdl-29485921
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.
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Texto completo: 1 Base de dados: MEDLINE Assunto principal: Hibridomas / Anticorpos Monoclonais / Especificidade de Anticorpos Idioma: En Ano de publicação: 2018 Tipo de documento: Article País de afiliação: Estados Unidos

Texto completo: 1 Base de dados: MEDLINE Assunto principal: Hibridomas / Anticorpos Monoclonais / Especificidade de Anticorpos Idioma: En Ano de publicação: 2018 Tipo de documento: Article País de afiliação: Estados Unidos