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  Seeing better through a MIST : evaluation of monoclonal recombinant antibody fragments on microarrays

Angenendt, P., Wilde, J., Kijanka, G., Baars, S., Cahill, D. J., Kreutzberger, J., et al. (2004). Seeing better through a MIST: evaluation of monoclonal recombinant antibody fragments on microarrays. Analytical Chemistry, 76(10), 2916-2921. doi:10.1021/ac035357a S0003-2700(03)05357-5.

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Genre: Journal Article
Alternative Title : Anal Chem

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 Creators:
Angenendt, Philipp1, Author              
Wilde, Jeannine1, Author              
Kijanka, Gregor, Author
Baars, Sabine, Author
Cahill, Dolores J., Author
Kreutzberger, Jürgen1, Author              
Lehrach, Hans1, Author              
Konthur, Zoltán2, Author              
Glökler, Jörn1, Author              
Affiliations:
1Dept. of Vertebrate Genomics (Head: Hans Lehrach), Max Planck Institute for Molecular Genetics, Max Planck Society, ou_1433550              
2In vitro Ligand Screening (Zoltán Konthur), Dept. of Vertebrate Genomics (Head: Hans Lehrach), Max Planck Institute for Molecular Genetics, Max Planck Society, ou_1479653              

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 Abstract: Automation is the key approach for genomewide and proteomewide screening of function and interaction. Especially for proteomics, antibody microarrays are a useful tool for massive parallel profiling of complex samples. To meet the requirements of antibody microarrays and to obtain a great variety of antibodies, new technologies such as phage display have partly replaced the classical hybridoma method. While the selection process for phage-displayed antibody fragments itself has been automated, the bottleneck was shifted further downstream to the identification of monoclonal binders obtained from the selections. Here, we present a new approach to reduce time, material, and waste to extend automation beyond the selection process by application of conventional microarray machinery. We were able to express recombinant antibody fragments in a single inoculation and expression step and subjected them without purification directly to an automated high-throughput screening procedure based on the multiple spotting technique (MIST). While obtaining comparable sensitivities to enzyme-linked immunosorbent assays, we minimized manual interaction steps and streamlined the technique to be accessible within the automated selection procedure.

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Language(s): eng - English
 Dates: 2004-10
 Publication Status: Published in print
 Pages: -
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 Table of Contents: -
 Rev. Type: -
 Identifiers: eDoc: 229481
DOI: 10.1021/ac035357a S0003-2700(03)05357-5
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Title: Analytical Chemistry
  Alternative Title : Anal Chem
Source Genre: Journal
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Pages: - Volume / Issue: 76 (10) Sequence Number: - Start / End Page: 2916 - 2921 Identifier: ISSN: 0003-2700