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  Evaluation of Recombinant Antibodies on Protein Microarrays Applying the Multiple Spotting Technique

Konthur, Z., & Wilde, J. (2010). Evaluation of Recombinant Antibodies on Protein Microarrays Applying the Multiple Spotting Technique. In R. Kontermann, & S. Dübel (Eds.), Antibody Engineering (pp. 447-460). Berlin et al: Springer-Verlag.

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 Creators:
Konthur, Zoltán1, Author              
Wilde, Jeannine2, Author              
Affiliations:
1In vitro Ligand Screening (Zoltán Konthur), Dept. of Vertebrate Genomics (Head: Hans Lehrach), Max Planck Institute for Molecular Genetics, Max Planck Society, ou_1479653              
2Dept. of Vertebrate Genomics (Head: Hans Lehrach), Max Planck Institute for Molecular Genetics, Max Planck Society, ou_1433550              

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 Abstract: The generation of recombinant antibodies by phage display in high-throughput demands fast downstream technologies and streamlined processes for the identification and initial characterisation of individual binders. Next to standard immunological methods such as enzyme-linked immunosorbent assays (ELISA) and Western-blot, protein microarrays offer a wide range of possibilities in the evaluation process of monoclonal binders. Here, we describe the application of a special protein microarray method – the multiple spotting technique (MIST) – for the simultaneous evaluation of hundreds of phage display derived soluble monoclonal antibody fragments on protein microarrays. The standard operating procedures provided include the expression of soluble antibody fragments in microtitre plates, the spotting protocols and data evaluation schemes. Additionally, we show the comparability of this protein microarray application to conventional ELISA on a recent target antigen in our semi-automated selection pipeline. Applying MIST allows to reduce time, material and waste, and extends automation beyond the selection process applying conventional microarray machinery.

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Language(s): eng - English
 Dates: 2010
 Publication Status: Published in print
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 Rev. Type: -
 Identifiers: eDoc: 556527
DOI: 10.1007/978-3-642-01147-4_34
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Title: Antibody Engineering
Source Genre: Book
 Creator(s):
Kontermann, Roland, Editor
Dübel, Stefan, Editor
Affiliations:
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Publ. Info: Berlin et al : Springer-Verlag
Pages: - Volume / Issue: - Sequence Number: - Start / End Page: 447 - 460 Identifier: ISBN: 978-3-642-01143-7

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Title: Springer Protocols
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