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          Institute: MPI für molekulare Genetik     Collection: Department of Vertebrate Genomics     Display Documents

ID: 556527.0, MPI für molekulare Genetik / Department of Vertebrate Genomics
Evaluation of Recombinant Antibodies on Protein Microarrays Applying the Multiple Spotting Technique
Authors:Konthur, Zoltán; Wilde, Jeannine
Place of Publication:Berlin et al
Date of Publication (YYYY-MM-DD):2010
Title of Book:Antibody Engineering
Start Page:447
End Page:460
Full Name of Book-Editor(s):Roland Kontermann und Stefan Dübel
Title of Series:Springer Protocols
Copyright:© Springer, Part of Springer Science+Business Media
Review Status:not specified
Audience:Experts Only
Abstract / Description: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.
External Publication Status:published
Document Type:InBook
Communicated by:Hans Lehrach
Affiliations:MPI für molekulare Genetik
Identifiers:ISBN:978-3-642-01143-7 [ID No:1]
DOI:10.1007/978-3-642-01147-4_34 [ID Nr:2]
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